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United States Patent Application 20180133297
Kind Code A1
Bae; Jooeun ;   et al. May 17, 2018

XBP1, CD138, AND CS1 PEPTIDES, PHARMACEUTICAL COMPOSITIONS THAT INCLUDE THE PEPTIDES, AND METHODS OF USING SUCH PEPTIDES AND COMPOSITIONS

Abstract

The disclosure features, inter alia, immunogenic XBP1-, CD138-, and CS1-derived peptides (and pharmaceutical compositions thereof). The peptides can be used in a variety of methods such as methods for inducing an immune response, methods for producing an antibody, and methods for treating a cancer (e.g., breast cancer, colon cancer, pancreatic cancer, a blood cancer, e.g., leukemia or a plasma cell disorder such as multiple myeloma or Waldenstrom's macroglobulinemia). The peptides (and pharmaceutical compositions comprising the peptides) can be used, e.g., in a method of treating a precancerous condition such as smoldering multiple myeloma. The peptides can also be included in MHC molecule multimer compositions and used in, e.g., methods for detecting a T cell in a population of cells.


Inventors: Bae; Jooeun; (West Roxbury, MA) ; Munshi; Nikhil; (Needham, MA) ; Anderson; Kenneth; (Wellesley, MA)
Applicant:
Name City State Country Type

DANA-FARBER CANCER INSTITUTE, INC.

Boston

MA

US
Family ID: 1000003125121
Appl. No.: 15/689159
Filed: August 29, 2017


Related U.S. Patent Documents

Application NumberFiling DatePatent Number
14440442May 4, 2015
PCT/US13/68582Nov 5, 2013
15689159
61790780Mar 15, 2013
61722446Nov 5, 2012

Current U.S. Class: 1/1
Current CPC Class: A61K 2039/55511 20130101; A61K 2039/505 20130101; A61K 45/06 20130101; A61K 39/39558 20130101; A61K 38/1709 20130101; A61K 39/39 20130101; A61K 39/0011 20130101; A61K 38/1774 20130101; A61K 38/193 20130101
International Class: A61K 39/00 20060101 A61K039/00; A61K 39/395 20060101 A61K039/395; A61K 38/17 20060101 A61K038/17; A61K 45/06 20060101 A61K045/06; A61K 39/39 20060101 A61K039/39; A61K 38/19 20060101 A61K038/19

Goverment Interests



STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This invention was made with government support under grant numbers P01CA078378, P01CA155258, and P50CA100707 awarded by The National Institute of Health. The government has certain rights in the invention.
Claims



1-24. (canceled)

25. A method for inducing an immune response in a subject having prostate cancer, the method comprising delivering to the subject a composition comprising a non spliced XBP1 peptide of 35 amino acids or less in length comprising the amino acid sequence of SEQ ID NO:6, a spliced XBP1 peptide of 35 amino acids or less in length and comprising the amino acid sequence of SEQ ID NO:10, and a CD138 peptide of 35 amino acids or less in length and comprising the amino acid sequence of SEQ ID NO:12.

26. The method of claim 25, further comprising, after delivering the composition to the subject, determining if an immune response occurred in the subject.

27. The method of claim 25 or 26, wherein the subject is a human.

28-31. (canceled)

32. The method of claim 25, comprising administering the composition in combination with an additional treatment selected from one or more of a chemotherapeutic agent, ionizing radiation, or an immunotherapy agent.

33. The method of claim 25, further comprising administering to the subject one or more immune stimulating agents and/or one or more immune modulating agents.

34. The method of claim 33, wherein the one or more immune stimulating agents is selected from an adjuvant comprising carboxymethylcellulose, polyinosinic-polycytidylic acid, and poly-L-lysine double-stranded RNA; an adjuvant comprising a water-and-oil emulsion; and an adjuvant comprising a protein.

35. The method of claim 33, wherein the one or more immune modulating agents is selected from an antibody which activates the immune system and an adjuvant.

36. A method for treating a subject having prostate cancer, the method comprising administering to the subject a composition comprising a non spliced XBP1 peptide of 35 amino acids or less in length comprising the amino acid sequence of SEQ ID NO:6, a spliced XBP1 peptide of 35 amino acids or less in length and comprising the amino acid sequence of SEQ ID NO:10, and a CD138 peptide of 35 amino acids or less in length and comprising the amino acid sequence of SEQ ID NO:12.

37-79. (canceled)

80. The method of claim 25, wherein the composition further comprises a CS-1 peptide of 35 amino acids or less in length comprising the amino acid sequence of SEQ ID NO:16.

81. The method of claim 35, wherein the adjuvant is a thalidomide derivative.

82. The method of claim 81, wherein the adjuvant is lenalidomide.

83. The method of claim 25, wherein the subject expresses HLA-A2.

84. The method of claim 36, wherein the composition further comprises a CS-1 peptide of 35 amino acids or less in length comprising the amino acid sequence of SEQ ID NO:16.

85. A method for inducing an immune response in a subject having multiple myeloma, breast cancer, colon cancer, pancreatic cancer, prostate cancer, or leukemia, the method comprising delivering to the subject a composition comprising a non spliced XBP1 peptide of 35 amino acids or less in length comprising the amino acid sequence of SEQ ID NO: 29, a spliced XBP1 peptide of 35 amino acids or less in length and comprising the amino acid sequence of SEQ ID NO: 30, and a CD138 peptide of 35 amino acids or less in length and comprising the amino acid sequence of SEQ ID NO: 31.

86. The method of claim 85, wherein the composition further comprises a CS-1 peptide of 35 amino acids or less in length comprising the amino acid sequence of SEQ ID NO: 32.

87. The method of claim 85, wherein the multiple myeloma is smoldering multiple myeloma.

88. The method of claim 85, wherein the subject expresses HLA-A24.

89. A method for treating a subject having multiple myeloma, breast cancer, colon cancer, pancreatic cancer, prostate cancer, or leukemia, the method comprising administering to the subject a composition comprising a non spliced XBP1 peptide of 35 amino acids or less in length comprising the amino acid sequence of SEQ ID NO: 29, a spliced XBP1 peptide of 35 amino acids or less in length and comprising the amino acid sequence of SEQ ID NO: 30, and a CD138 peptide of 35 amino acids or less in length and comprising the amino acid sequence of SEQ ID NO: 31.

90. The method of claim 89, wherein the composition further comprises a CS-1 peptide of 35 amino acids or less in length comprising the amino acid sequence of SEQ ID NO: 32.

91. The method of claim 89, wherein the subject expresses HLA-A24.
Description



CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation of Ser. No. 14/440,442, filed Nov. 5, 2013, which is a U.S. National Phase Application under 35 U.S.C. .sctn. 371 of International Application No. PCT/US2013/068582, filed Nov. 5, 2013, which claims priority to U.S. Ser. No. 61/722,446 filed Nov. 5, 2012, and U.S. Ser. No. 61/790,780 filed Mar. 15, 2013 the entire contents of which are incorporated herein by reference.

SEQUENCE LISTING

[0003] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format. Said ASCII copy, created on Dec. 9, 2013, is named 02017-7001WO_SL.txt and is 117,979 bytes in size.

BACKGROUND

[0004] Several types of vaccines have been developed for the prevention of infectious diseases including attenuated microorganisms, recombinant proteins and DNA vaccines. Recently, research has been carried out on the development of vaccine immunotherapy to treat cancer patients.

SUMMARY

[0005] The present disclosure relates to immunogenic peptides that bind to MHC class 1 molecules such as HLA-A molecules. It was found that peptides from X-Box Protein 1 (XBP1)-, CD138-, and CD2 Subset 1 (CS1) are immunogenic and are useful, e.g., to induce an immune response against various cancer cells. In some embodiments the peptides possess elevated affinity for HLA-A molecules, elevated stability within the peptide binding cleft of HLA-A, and the ability, when expressed on the surface of cell (e.g., a cancer cell) in the context of an MHC molecule, to induce the activation and proliferation of T cells (e.g., effector memory T cells and/or central memory T cells).

[0006] In addition, it was found that combinations of these peptides can induce a broad spectrum of immune responses against the target antigens and this broad spectrum response can overcome major therapeutic hurdles including, inter alia, the heterogeneity of tumor associated antigen expression, frequent mutations of specific antigens and the variability of the human T-cell repertoire among individuals. Thus, administration of various combinations of these peptides, e.g., combined in a pharmaceutical composition, may provide an enhanced immune response against various cancers.

[0007] It will be evident from the following description that the peptides (and pharmaceutical compositions thereof) can be used in a variety of applications such as methods for inducing an immune response, methods for activating a T cell (e.g., including effectory memory T cells and/or central memory T cells), methods for producing an antibody, and methods for treating, e.g., a cancer (e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML), multiple myeloma, Waldenstrom's Macroglobulinemia and precancerous conditions, e.g., smoldering multiple myeloma.

[0008] In one aspect, the disclosure features peptides, e.g., XBP1 peptides, CD138 peptides and CS-1 peptides, that have affinity for multiple MHC molecules, e.g., HLA-A molecules such as HLA-A2 and HLA-A24, elevated stability within the peptide binding cleft of multiple MHC molecules, e.g., HLA-A2 and HLA-A24, and the ability, when expressed on the surface of cell (e.g., a cancer cell) in the context of an MHC molecule, e.g., HLA-A2 or HLA-A24, to induce the activation and proliferation of T cells including, e.g., effector memory T cells and/or central memory T cells).

[0009] It will be evident from the following description that the peptides (and pharmaceutical compositions thereof) can be used in a variety of applications such as methods for inducing an immune response, methods for activating a T cell (e.g., effector memory T cells and/or central memory T cells), methods for producing an antibody, and methods for treating, e.g., a cancer (e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML), multiple myeloma and precancerous conditions, e.g., smoldering multiple myeloma.

[0010] In one aspect, the disclosure features an isolated peptide comprising an amino acid sequence that is at least 66 (e.g., at least 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99) % identical to any one of SEQ ID NOS: 51-536. The peptide can bind to a major histocompatibility complex (MHC) molecule such as an MHC class I or class II molecule. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 51-536 or an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to any of SEQ ID NOs: 51-536. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 51-536. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, or one substitution(s) of any one of SEQ ID NOs: 51-536. The substitutions can be conservative or non-conservative. In one embodiment, the peptide is used to treat a subject having or at risk of having cancer, e.g., a cancer described herein, e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma.

[0011] In one embodiment, the peptide is used to treat a subject having a precancerous condition, e.g., smoldering multiple myeloma.

[0012] In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck (e.g, nasopharyngeal cancer). In one embodiment, the cancer is breast cancer (e.g., invasive lobular, invasive ductal, mixed lobular and ductal, intraductal cribriform, invasive ductal and lobular or invasive carcinoma). In one embodiment, the cancer is a colon adenocarcinoma (e.g., mucinous adenocarcinoma).

[0013] In one embodiment, the peptide consists of an amino acid sequence that is at least 66 (e.g., at least 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99) % identical to any one of SEQ ID NOS: 51-536.

[0014] In one embodiment, the peptide consists of an amino acid sequence with three, two or one substitution of any one of SEQ ID NOS:51-536. In one embodiment, the peptide consists of an amino acid sequence of any one of SEQ ID NOS: 51-536.

[0015] In one embodiment, the peptide is a non-spliced XBP1 peptide of Group C (see, e.g., Table 3), e.g., a non-spliced XBP-1 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 51-206 or a non-spliced XBP1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 51-206. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 51-206 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 51-206. In one embodiment, the non-spliced XBP1 peptide of Group C is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of any one of SEQ ID NOS: 51-206. The substitutions can be conservative or non-conservative. In one embodiment, the non-spliced XBP1 peptide of Group C consists of an amino acid sequence of any one of SEQ ID NOS: 51-206.

[0016] In one embodiment, the peptide is a CD138 peptide of Group C, e.g., a CD138 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 207-371 or a CD138 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with an amino acid sequence of any one of SEQ ID NOS: 207-371. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 207-371 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 207-371. In one embodiment, the CD138 peptide of Group C is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of an amino acid sequence of any one of SEQ ID NOS: 207-371. The substitutions can be conservative or non-conservative. In one embodiment, the CD138 peptide of Group C consists of an amino acid sequence of any one of SEQ ID NOS: 207-371.

[0017] In one embodiment, the peptide is a CS-1 peptide of Group C, e.g., a CS-1 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 372-536 or a CS-1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with an amino acid sequence of any one of SEQ ID NOS: 372-536. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 372-536 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 372-536. In one embodiment, the CS-1 peptide of Group C is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of any one of SEQ ID NOS: 372-536. The substitutions can be conservative or non-conservative. In one embodiment, the CS-1 peptide of Group C consists of an amino acid sequence of any one of SEQ ID NOS: 372-536.

[0018] In one embodiment, the peptide of Group C, e.g., the XBP peptide, the CD138 peptide and/or the CS-1 peptide, is used to treat a subject having or at risk of having cancer, e.g., a cancer described herein, e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML. In one embodiment, the peptide of Group C is used to treat a subject having a precancerous condition, e.g., smoldering multiple myeloma. In an embodiment, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck (e.g, nasopharyngeal cancer).

[0019] In some embodiments, any of the isolated peptides described herein can bind to a major histocompatibility complex (MHC) molecule (e.g., an MHC class I or class II molecule). The MHC molecule can be, e.g., a human MHC molecule. The MHC molecule can be, e.g., an HLA-A molecule, an HLA-B molecule and/or an HLA-C molecule. Preferably, the MHC molecule is one or more HLA-A molecule (e.g., HLA-A1, HLA-A2, HLA-A3 and HLA-A24).

[0020] In one aspect, the disclosure features immunogenic X-Box Protein 1 (XBP1)-, CD138-, and CD2 Subset 1 (CS1)-derived peptides, e.g., which possess elevated affinity for HLA-A2 molecules, elevated stability within the peptide binding cleft of HLA-A2, and the ability, when expressed on the surface of cell (e.g., a cancer cell) in the context of an MHC molecule, to induce the activation and proliferation of T cells (e.g., effector memory T cells and/or central memory T cells). For example, all or a subset of these peptides are expressed on the surface of various cancer cells, including multiple myeloma cells, and specifically smoldering multiple myeloma cells, colon cancer cells, breast cancer cells, pancreatic cancer cells, prostate cancer cells and leukemia, e.g., acute myeloid leukemia (AML) cells, in the context of an MHC molecule, and the presence of these peptides induces activation and proliferation of T cells against these and other cancers.

[0021] In addition, it was found that combinations of these peptides can induce a broad spectrum of immune responses against the target antigens and this broad spectrum response can overcome major therapeutic hurdles including, inter alia, the heterogeneity of tumor associated antigen expression, frequent mutations of specific antigens and the variability of the human T-cell repertoire among individuals. Thus, administration of combinations of these peptides, e.g., combined in a pharmaceutical composition, may provide an enhanced immune response against various cancers.

[0022] It will be evident from the following description that the peptides (and pharmaceutical compositions thereof) can be used in a variety of applications such as methods for inducing an immune response, methods for activating a T cell (e.g., effector memory T cells and/or central memory T cells), methods for producing an antibody, and methods for treating, e.g., a cancer (e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML), multiple myeloma and precancerous conditions, e.g., smoldering multiple myeloma.

[0023] In one aspect, the disclosure features an isolated peptide comprising an amino acid sequence that is at least 66 (e.g., at least 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99) % identical to any one of SEQ ID NOS:1-18. The peptide can bind to a major histocompatibility complex (MHC) molecule such as an MHC class I or class II molecule. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS:1-18, or an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence of SEQ ID NOS:1-18. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS:1-18. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, or one substitution(s) of an amino acid sequence of any one of SEQ ID NOS:1-18. The substitutions can be conservative or non-conservative. In one embodiment, the peptide is used to treat a subject having or at risk of having cancer, e.g., a cancer described herein, e.g., breast cancer (e.g., invasive lobular, invasive ductal, mixed lobular and ductal, intraductal cribriform, invasive lobular and ductal, invasive carcinoma), colon cancer (e.g., adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma.

[0024] In one embodiment, the peptide is used to treat a subject having a precancerous condition, e.g., smoldering multiple myeloma. In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck.

[0025] In one embodiment, the peptide consists of an amino acid sequence that is at least 66 (e.g., at least 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99) % identical to any one of SEQ ID NOS:1-18. In one embodiment, the peptide consists of any of amino acid sequences of SEQ ID NOS;1-18 with three, two or one substitution. In one embodiment, the peptide consists of an amino acid sequence of any one of SEQ ID NOS:1-18.

[0026] In one embodiment, the peptide is a non-spliced XBP1 peptide of Group A (see, e.g., Table 1), e.g., a non-spliced XBP-1 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 1-6, a non-spliced XBP1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with an amino acid sequence of any one of SEQ ID NOS:1-6. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS:1-6 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with an amino acid sequence of any one of SEQ ID NOS:1-6. In one embodiment, the non-spliced XBP1 peptide of Group A is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of an amino acid sequence of any one of SEQ ID NOS:1-6. The substitutions can be conservative or non-conservative. In one embodiment, the non-spliced XBP1 peptide of Group A consists of an amino acid sequence of any one of SEQ ID NOS:1-6.

[0027] In one embodiment, the peptide is a spliced XBP1 peptide of Group A, e.g., a spliced XBP-1 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 7-10, a spliced XBP1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with an amino acid sequence of any one of SEQ ID NOS:7-10. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS:7-10 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with an amino acid sequence of any one of SEQ ID NOS:7-10. In one embodiment, the spliced XBP1 peptide of Group A is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of an amino acid sequence of any one of SEQ ID NOS:7-10. The substitutions can be conservative or non-conservative. In one embodiment, the spliced XBP1 peptide of Group A consists of an amino acid sequence of any one of SEQ ID NOS:7-10.

[0028] In one embodiment, the peptide is a CD138 peptide of Group A, e.g., a CD138 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 11-14, a CD138 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with an amino acid sequence of any one of SEQ ID NOS:11-14. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS:11-14 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with an amino acid sequence of any one of SEQ ID NOS:11-14. In one embodiment, the CD138 peptide of Group A is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of an amino acid sequence of any one of SEQ ID NOS:11-14. The substitutions can be conservative or non-conservative. In one embodiment, the CD138 peptide of Group A consists of an amino acid sequence of any one of SEQ ID NOS:11-14.

[0029] In one embodiment, the peptide is a CS-1 peptide of Group A, e.g., a CS-1 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 15-18, a CS-1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with an amino acid sequence of any one of SEQ ID NOS:15-18. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS:15-18 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with an amino acid sequence of any one of SEQ ID NOS:15-18. In one embodiment, the CS-1 peptide of Group A is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of an amino acid sequence of any one of SEQ ID NOS:15-18. The substitutions can be conservative or non-conservative. In one embodiment, the CS-1 peptide of Group A consists of an amino acid sequence of any one of SEQ ID NOS:15-18.

[0030] In one embodiment, the peptide of Group A, e.g., the non-spliced XBP1 peptide, the spliced XBP1 peptide, the CD138 peptide and/or the CS-1 peptide, is used to treat a subject having or at risk of having cancer, e.g., a cancer described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML. In one embodiment, the peptide of Group A, e.g., the non-spliced XBP1 peptide, the spliced XBP1 peptide, the CD138 peptide and/or the CS-1 peptide, is used to treat a subject having a precancerous condition, e.g., smoldering multiple myeloma. In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck.

[0031] In some embodiments, any of the isolated peptides described herein can bind to a major histocompatibility complex (MHC) molecule (e.g., an MHC class I or class II molecule). The MHC molecule can be, e.g., an HLA-A2 molecule. The MHC molecule can be, e.g., a human MHC molecule.

[0032] In another aspect, the disclosure features immunogenic X-Box Protein 1 (XBP1)-, CD138-, and CD2 Subset 1 (CS1)-derived peptides, e.g., which possess elevated affinity for HLA-A24 molecules, elevated stability within the peptide binding cleft of HLA-A24, and the ability, when expressed on the surface of cell (e.g., a cancer cell) in the context of an MHC molecule, to induce the activation and proliferation of T cells (e.g., effector memory T cells and/or central memory T cells). For example, all or a subset of these peptides are expressed on the surface of various cancer cells, including multiple myeloma cells, colon cancer cells, breast cancer cells, pancreatic cancer cells, prostate cancer cells and leukemia, e.g., acute myeloid leukemia (AML) cells, in the context of an MHC molecule, and the presence of these peptides induces activation and proliferation of T cells against these and other cancers.

[0033] It will be evident from the following description that the peptides (and pharmaceutical compositions thereof) can be used in a variety of applications such as methods for inducing an immune response, methods for activating a T cell (e.g., effector memory T cells and/or central memory T cells), methods for producing an antibody, and methods for treating, e.g., a cancer (e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML), multiple myeloma and precancerous conditions, e.g., smoldering multiple myeloma.

[0034] In one aspect, the disclosure features an isolated peptide comprising an amino acid sequence that is at least 66 (e.g., at least 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99) % identical to any one of SEQ ID NOS: 29-50. The peptide can bind to a major histocompatibility complex (MHC) molecule such as an MHC class I or class II molecule. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 29-50, or an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence of SEQ ID NOS: 29-50. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 29-50. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, or one substitution(s) of an amino acid sequence of any one of SEQ ID NOS: 29-50. The substitutions can be conservative or non-conservative. In one embodiment, the peptide is used to treat a subject having or at risk of having cancer, e.g., a cancer described herein, e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer (e.g., invasive lobular, invasive ductal, mixed lobular and ductal, intraductal cribriform, invasive lobular and ductal, invasive carcinoma), colon cancer (e.g., colon adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma.

[0035] In one embodiment, the peptide is used to treat a subject having a precancerous condition, e.g., smoldering multiple myeloma. In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck (e.g., nasopharyngeal cancer).

[0036] In one embodiment, the peptide consists of an amino acid sequence that is at least 66 (e.g., at least 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99) % identical to any one of SEQ ID NOS: 29-50. In one embodiment, the peptide consists of an amino acid sequence of any one of SEQ ID NOS:29-50 with three, two or one substitution. In one embodiment, the peptide consists of an amino acid sequence of any one of SEQ ID NOS: 29-50.

[0037] In one embodiment, the peptide is a non-spliced XBP1 peptide from Group B (see Table 2), e.g., a non-spliced XBP-1 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 29 and 33-37 or a non-spliced XBP1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 29 and 33-37. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 29 and 33-37 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 29 and 33-37. In one embodiment, the non-spliced XBP1 peptide from Group B is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of any one of SEQ ID NOS: 29 and 33-37. The substitutions can be conservative or non-conservative. In one embodiment, the non-spliced XBP1 peptide is from amino acid sequence SEQ ID NO:19 and comprises the amino acid sequence of SEQ ID NO:29 and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3) amino acids C-terminal to SEQ ID NO:29 in SEQ ID NO:19. In one embodiment, the non-spliced XBP1 peptide is from amino acid sequence SEQ ID NO:19 and comprises the amino acid sequence of SEQ ID NO:33 and 1, 2, 3, 4, 5, or more (e.g., 1, or 2) amino acids N-terminal to SEQ ID NO:33 in SEQ ID NO:19 and/or 1, 2, 3 or moire (e.g., 1) amino acids C-terminal to SEQ ID NO:33 in SEQ ID NO:19. In one embodiment, the non-spliced XBP1 peptide is from amino acid sequence SEQ ID NO:19 and comprises the amino acid sequence of SEQ ID NO:36 and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3, or 4) amino acids N-terminal to SEQ ID NO:36 in SEQ ID NO:19 and/or 1, 2, 3, 5, 6 or more (e.g., 1, 2, 3, 4, 5 or 6) amino acids C-terminal to SEQ ID NO:36 in SEQ ID NO:19. In one embodiment, the non-spliced XBP1 peptide is from amino acid sequence SEQ ID NO:19 and comprises the amino acid sequence of any one of SEQ ID NOS: 34, 35 or 37 and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3) amino acids N-terminal and/or C-terminal to any one of SEQ ID NOS: 34, 35 or 37 in SEQ ID NO:19. In one embodiment, the non-spliced XBP1 peptide from Group B consists of an amino acid sequence of any one of SEQ ID NOS: 29 and 33-37.

[0038] In one embodiment, the peptide is a spliced XBP1 peptide from Group B, e.g., a spliced XBP-1 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 30, 38 and 39 or a spliced XBP1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 30, 38 and 39. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 30, 38 and 39 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 30, 38 and 39. In one embodiment, the spliced XBP1 peptide from Group B is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of any one of SEQ ID NOS: 30, 38 and 39. The substitutions can be conservative or non-conservative. In one embodiment, the spliced XBP1 peptide is from amino acid sequence SEQ ID NO:20 and comprises the amino acid sequence of any one of SEQ ID NOS: 30, 38 and 39 and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3) amino acids N-terminal and/or C-terminal to any one of SEQ ID NOS: 30, 38, and 39 in SEQ ID NO:20 In one embodiment, the spliced XBP1 peptide from Group B consists of an amino acid sequence of any one of SEQ ID NOS: 30, 38 and 39.

[0039] In one embodiment, the peptide is a CD138 peptide from Group B, e.g., a CD138 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 31 or a CD138 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 31 and 40-45. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 31 and 40-45 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 31 and 40-45. In one embodiment, the CD138 peptide from Group B is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of any one of SEQ ID NOS: 31 and 40-45. The substitutions can be conservative or non-conservative. In one embodiment, the CD138 peptide is from amino acid sequence SEQ ID NO:21 and comprises the amino acid sequence of SEQ ID NO:31 and 1, 2, 3, 4, 5, 6, 7, 8 or more (e.g., 1, 2, 3, 4) amino acids N-terminal to SEQ ID NO:31 in SEQ ID NO:21. In one embodiment, the CD138 peptide is from amino acid sequence SEQ ID NO:21 and comprises the amino acid sequence of SEQ ID NO:42 or 44 and 1, 2, 3, 4, 5, or more (e.g., 1, 2, 3, 4) amino acids N-terminal to SEQ ID NO:42 or 44 in SEQ ID NO:21 and/or 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3, 4, 5, 6) amino acids C-terminal to SEQ ID NO:42 or 44 in SEQ ID NO:21. In one embodiment, the CD138 peptide is from amino acid sequence SEQ ID NO:21 and comprises the amino acid sequence of SEQ ID NO:45 and 1, 2, 3, 4, 5, 6, 7, 8, or more (e.g., 1, 2, 3, or 4) amino acids N-terminal to SEQ ID NO:45 in SEQ ID NO:21 and/or 1, 2, 3, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3, 4, 5 or 6) amino acids C-terminal to SEQ ID NO:45 in SEQ ID NO:21. In one embodiment, the CD138 peptide is from amino acid sequence SEQ ID NO:21 and comprises the amino acid sequence of any one of SEQ ID NOS: 40, 41 or 43 and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3) amino acids N-terminal and/or C-terminal to any one of SEQ ID NOS: 40, 41 or 43 in SEQ ID NO:21. In one embodiment, the CD138 peptide from Group B consists of an amino acid sequence of any one of SEQ ID NOS: 31 and 40-45.

[0040] In one embodiment, the peptide is a CS-1 peptide from Group B, e.g., a CS-1 peptide comprising an amino acid sequence of any one of SEQ ID NOS: 32 and 46-50 or a CS-1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 32 and 46-50. In one embodiment, the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 32 and 46-50 or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with any one of SEQ ID NOS: 32 and 46-50. In one embodiment, the CS-1 peptide from Group B is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and the peptide comprises an amino acid sequence with three, two, one substitutions of any one of SEQ ID NOS:

[0041] 32 and 46-50. The substitutions can be conservative or non-conservative. In one embodiment, the CS-1 peptide is from amino acid sequence SEQ ID NO:22 and comprises the amino acid sequence of SEQ ID NOS: 32 and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3) amino acids C-terminal to SEQ ID NOS:32 in SEQ ID NO:22. In one embodiment, the CS-1 peptide is from amino acid sequence SEQ ID NO:22 and comprises the amino acid sequence of any one of SEQ ID NOS: 46-50 and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3) amino acids N-terminal and/or C-terminal to any one of SEQ ID NOS: 46-50 in SEQ ID NO:22. In one embodiment, the CS-1 peptide from Group B consists of an amino acid sequence of any one of SEQ ID NOS: 32 and 46-50.

[0042] In one embodiment, the peptide of Group B, e.g., a non-spliced XBP1 peptide, a spliced XBP1 peptide, a CD138 peptide and/or a CS-1 peptide, is used to treat a subject having or at risk of having cancer, e.g., a cancer described herein, e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML. In one embodiment, the peptide of Group B, e.g., a non-spliced XBP1 peptide, a spliced XBP1 peptide, a CD138 peptide and/or a CS-1 peptide, is used to treat a subject having a precancerous condition, e.g., smoldering multiple myeloma. In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck (e.g, nasopharyngeal cancer).

[0043] In some embodiments, any of the isolated peptides described herein can, in association with a major histocompatibility complex (MHC) molecule, be recognized by an antigen specific T cell receptor on a T cell.

[0044] In another aspect, the disclosure features a fusion protein that comprises a first amino acid sequence consisting of a peptide described herein, e.g., a non-spliced XBP1 peptide from Group A, Group B or Group C, a spliced XBP1 peptide from Group A or Group B, a CD138 peptide from Group A, Group B or Group C and/or a CS-1 peptide from Group A, Group B or Group C, e.g., described herein; and a second amino acid sequence that is heterologous to the first amino acid sequence.

[0045] In some embodiments, the second amino acid sequence can comprise, or be, a targeting polypeptide, an immune stimulatory molecule, an immunoglobulin or antigen-binding fragment thereof, an Fc receptor-binding region of an immunoglobulin molecule, or a carrier polypeptide. The targeting polypeptide can be, e.g., one that targets the isolated peptide to an antigen presenting cell (e.g., a dendritic cell, a macrophage, a monocyte, or a B cell). The immune stimulatory molecule can be, e.g., a cytokine or a T helper epitope. The immunoglobulin can be, e.g., a single chain Fv immunoglobulin fragment or an entire immunoglobulin molecule. The carrier polypeptide can comprise, or be, a KLH (keyhole limpet hemocyanin) polypeptide, or an albumin polypeptide.

[0046] In some embodiments, any of the isolated peptides described herein can contain a linker sequence. The linker sequence can directly or indirectly connect a first amino acid sequence to a second amino acid sequence. The linker sequence can comprise, or consist of, one or more amino acids, e.g., at least one, two, three, four, five, six, seven, eight, nine or ten amino acids. In one embodiment, the linker can comprise, or consist of, at least one (e.g., one, two, three, four, five, six, seven, eight, nine, or 10 or more) protease cleavage site.

[0047] In some embodiments, the second amino acid sequence can be amino terminal or carboxy terminal to the first amino acid sequence.

[0048] In some embodiments, any of the isolated peptides or fusion proteins described herein can be detectably labeled. The detectable label can be selected from the group consisting of luminescent labels, fluorescent labels, radioactive labels, and enzymatic labels.

[0049] In yet another aspect, the disclosure features: (i) an isolated nucleic acid encoding any of the isolated peptides described herein; (ii) a vector comprising the isolated nucleic acid of (i); or (iii) a cultured cell comprising the vector of (ii). The vector can be operably linked to an expression control sequence. The cultured cell can be a prokaryotic cell or eukaryotic cell. The cultured cell can be, e.g., a fungal cell, a plant cell, or an animal cell (e.g., a nematode cell, an insect cell, a bird cell, a fish cell, or a mammalian cell (e.g., a human cell)). The cultured cell can be an immune cell such as any of the immune cells described herein.

[0050] In another aspect, the disclosure features a method of producing a peptide. The method involves the step of culturing any of the cultured cells described herein under conditions that permit the expression of the peptide. The method can also include the step of isolating the peptide from the cell or from the medium in which the cell was cultured.

[0051] In another aspect, the disclosure features a pharmaceutical composition comprising one or more of any of the isolated peptides (or fusion proteins) described herein and a pharmaceutically acceptable carrier. In one embodiment, the composition comprises at least two peptides, e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more of the peptides described herein. For example, in one embodiment, the composition comprises at least two, three or four of the peptides described herein.

[0052] In one embodiment, the composition comprises at least two peptides. For example, the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group A, and a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group A; the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group A, and a CD138 peptide, e.g., a CD138 peptide from Group A; the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group A, and a CS-1 peptide, e.g., a CS-1 peptide from Group A; the composition comprises a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group A, and a CD138 peptide, e.g., a CD138 peptide from Group A; the composition comprises a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group A, and a CS-1 peptide, e.g., a CS-1 peptide from Group A; the composition comprises a CD138 peptide, e.g., a CD138 peptide from Group A, and a CS-1 peptide, e.g., a CS-1 peptide from Group A.

[0053] In one embodiment, the composition comprises at least three peptides. For example, the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide described from Group A, a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group A, and a CD138 peptide, e.g., a CD138 peptide from Group A; the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group A, a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group A and a CS-1 peptide, e.g., a CS-1 peptide from Group A; the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group A, a CD138 peptide, e.g., a CD138 peptide from Group A and a CS-1 peptide, e.g., a CS-1 peptide from Group A; the composition comprises a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group A, a CD138 peptide, e.g., a CD138 peptide from Group A and a CS-1 peptide, e.g., a CS-1 peptide from Group A. In one embodiment, the composition comprises at least three peptides, e.g., a non-spliced XBP1 peptide (e.g., a non-spliced XBP1 peptide from Group A), a spliced XBP1 peptide (e.g., a spliced XBP-1 peptide from Group A), and a CD138 peptide (e.g., a CD138 peptide from Group A).

[0054] In one embodiment, the composition comprises four peptides, e.g., the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group A, a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group A, a CD138 peptide, e.g., a CD138 peptide from Group A, and a CS-1 peptide, e.g., a CS-1 peptide from Group A.

[0055] In one embodiment, the composition comprises a non-spliced XBP1 peptide from Group A that is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any of SEQ ID NOS:1-6, e.g., SEQ ID NO:6. In one embodiment, the composition comprises a spliced XBP1 peptide from Group A that is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any of SEQ ID NOS:7-10, e.g., SEQ ID NO:10. In one embodiment, the composition comprises a CD138 peptide from Group A that is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any of SEQ ID NOS:11-14, e.g., SEQ ID NO:12. In one embodiment, the composition comprises a CS-1 peptide from Group A that is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any of SEQ ID NOS:15-18, e.g., SEQ ID NO:16.

[0056] In one embodiment, the composition comprises four peptides and the four peptides are a peptide from Group A that comprises (e.g., consists of) the amino acid sequence of SEQ ID NO:6, a peptide from Group A that comprises (e.g., consists of) the amino acid sequence of SEQ ID NO:10, a peptide from Group A that comprises (e.g., consists of) the amino acid sequence of SEQ ID NO:12, and a peptide from Group A that comprises (e.g., consists of) the amino acid sequence of SEQ ID NO:16.

[0057] In one embodiment, the composition comprises at least two peptides. For example, the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group B, and a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group B; the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group B, and a CD138 peptide, e.g., a CD138 peptide from Group B; the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group B, and a CS-1 peptide, e.g., a CS-1 peptide from Group B; the composition comprises a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group B, and a CD138 peptide, e.g., a CD138 peptide from Group B; the composition comprises a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group B, and a CS-1 peptide, e.g., a CS-1 peptide from Group B; the composition comprises a CD138 peptide, e.g., a CD138 peptide from Group B, and a CS-1 peptide, e.g., a CS-1 peptide from Group B.

[0058] In one embodiment, the composition comprises at least three peptides. For example, the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide described from Group B, a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group B, and a CD138 peptide, e.g., a CD138 peptide from Group B; the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group B, a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group B and a CS-1 peptide, e.g., a CS-1 peptide from Group B; the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group B, a CD138 peptide, e.g., a CD138 peptide from Group B and a CS-1 peptide, e.g., a CS-1 peptide from Group B; the composition comprises a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group B, a CD138 peptide, e.g., a CD138 peptide from Group B and a CS-1 peptide, e.g., a CS-1 peptide from Group B. In one embodiment, the composition comprises at least three peptides, e.g., a non-spliced XBP1 peptide (e.g., a non-spliced XBP1 peptide from Group B), a spliced XBP1 peptide (e.g., a spliced XBP-1 peptide from Group B), and a CD138 peptide (e.g., a CD138 peptide from Group B).

[0059] In one embodiment, the composition comprises four peptides, e.g., the composition comprises a non-spliced XBP1 peptide, e.g., a non-spliced XBP1 peptide from Group B, a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group B, a CD138 peptide, e.g., a CD138 peptide from Group B, and a CS-1 peptide, e.g., a CS-1 peptide from Group B.

[0060] In one embodiment, the composition comprises a non-spliced XBP1 peptide from Group B that is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 29 and 33-37. In one embodiment, the composition comprises a spliced XBP1 peptide from Group B that is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 30, 38 and 39. In one embodiment, the composition comprises a CD138 peptide from Group B that is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 31 and 40-45. In one embodiment, the composition comprises a CS-1 peptide from Group B that is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprises an amino acid sequence of any one of SEQ ID NOS: 32 and 46-50.

[0061] In one embodiment, the composition comprises four peptides and the four peptides are a peptide from Group B that comprises (e.g., consists of) the amino acid sequence of any one of SEQ ID NOS: 29 and 33-37, a peptide from Group B that comprises (e.g., consists of) the amino acid sequence of any one of SEQ ID NOS: 30, 38 and 39, a peptide from Group B that comprises (e.g., consists of) the amino acid sequence of any one of SEQ ID NOS: 31 and 40-45, and a peptide from Group B that comprises (e.g., consists of) the amino acid sequence of any one of SEQ ID NOS: 32 and 46-50.

[0062] In one embodiment, the composition comprises peptides from Group A and Group B. For example, the composition comprises 1, 2, 3, 4 or more peptides from Group A and 1, 2, 3, 4, or more peptides from Group B.

[0063] The composition can also include, e.g., one or more additional agents, e.g., one or more therapeutic agents, diagnostic agents, or prophylactic agents, or immune stimulating or modulating agents. Immune stimulating agents include, but are not limited to, e.g., a T helper epitope, an altered peptide ligand, an adjuvant, or any other immune stimulating agent described herein. The T helper epitope can be, e.g., a PADRE sequence or a universal Tetanus Toxoid T helper (TT Th) epitope. The adjuvant can be selected from the group consisting of Freund's complete adjuvant, Freund's incomplete adjuvant, alum, a ligand for a Toll receptor, saponin (e.g., QS21), RIBI, cholera toxin (CT), E. coli heat labile toxin (LT), mutant CT (MCT), mutant E. coli heat labile toxin (MLT), an adjuvant comprising carboxymethylcellulose, polyinosinic-polycytidylic acid, and poly-L-lysine double-stranded RNA (e.g., poly IC-LC, e.g., hiltonol), an adjuvant comprising a water-and-oil emulsion (e.g., montanide), and an adjuvant comprising a protein (e.g., cytokines, complements, GCSF, GM-CSF). In one embodiment, the immune stimulating agent is an adjuvant comprising carboxymethylcellulose, polyinosinic-polycytidylic acid, and poly-L-lysine double-stranded RNA, e.g., poly IC-LC, e.g., hiltonol. In one embodiment the adjuvant is a water-and-oil emulsion, e.g., montanide. In one embodiment, the adjuvant is a protein, e.g., a cytokine, a complement, GCSF, GM-CSF. In one embodiment, the immune modulating agent can be a protein, e.g., an antibody which modulates the immune system. For example, an antibody which modulates the immune system can be an anti-CTLA4 antibody, e.g., ipilimumab or tremelimumab, or an anti-PD-1 antibody, or anti-PDL-1 antibody. In one embodiment, the immune modulating agent can be a small molecule adjuvant, e.g., thalidomide or a thalidomide derivative, e.g., lenalidomide.

[0064] The composition may also include an immunogenic peptide other than one disclosed above, e.g., an immunogenic peptide from WT1 or a derivative thereof. Exemplary WT1 peptides are described in U.S. Pat. No. 7,598,221, the contents of which is incorporated herein by reference. In one embodiment, the composition comprises one or more immunogenic peptide from WT1 or a derivative thereof, e.g., selected from one or more of: a WT1 class 1 epitope; a peptide comprising (or consisting of) RMFPNAPYL (SEQ ID NO: 538) (WT1 126-134); a peptide comprising (or consisting of) YMFPNAPYL (SEQ ID NO: 539); a peptide comprising (or consisting of) RSDELVRHHNMHQRNMTKL (SEQ ID NO: 540) (WT1 427-445); a peptide comprising (or consisting of) PGCNKRYFKLSHLQMHSRKHTG (SEQ ID NO: 541) (WT1 331-352); a peptide comprising (or consisting of) SGQARMFPNAPYLPSCLES (SEQ ID NO: 542) (WT1 122-140); and a peptide comprising (or consisting of) SGQAYMFPNAPYLPSCLES (SEQ ID NO: 543). Other immunogenic peptides include, but are not limited to, an immunogenic peptide from MUC1, an immunogenic peptide from gp100, an immunogenic peptide from TRP-2, an immunogenic peptide from MAGI, an immunogenic peptide from NY-ESO1, an immunogenic peptide from HER-2; and an immunogenic peptide from AIM2.

[0065] In one embodiment, the composition described herein is used to treat a subject having or at risk of having cancer, e.g., a cancer described herein, e.g., breast cancer (e.g., invasive lobular, invasive ductal, mixed lobular and ductal, intraductal cribriform, invasive lobular and ductal, invasive carcinoma), colon cancer (e.g., colon adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma. In one embodiment, the composition described herein is used to treat a subject having a precancerous condition, e.g., smoldering multiple myeloma. In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck.

[0066] In yet another aspect, the disclosure features a kit comprising: (i) one or more of any of the isolated peptides from Group A, from Group B and/or Group C; and instructions for administering the peptide to a subject, e.g., a subject having cancer, e.g., a cancer described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma or a subject having a precancerous condition, e.g., smoldering multiple myeloma; (ii) a composition described herein and instructions for administering the peptide to a subject, e.g., a subject having cancer, e.g., a cancer described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma or a subject having a precancerous condition, e.g., smoldering multiple myeloma; and/or (iii) one or more of the isolated nucleic acids encoding the isolated peptides, one or more vectors containing the isolated nucleic acids, or one or more cultured cells containing the vectors, and instructions for producing the isolated peptides. In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck.

[0067] In some embodiments, the kits can also include, e.g., one or more pharmaceutically acceptable carriers, one or more immune stimulating or modulating agents, or one or more therapeutic agents, diagnostic agents, or prophylactic agents. In one embodiment, the immune stimulating agent is an immune stimulating agent described herein. The one or more immune stimulating agents can be selected from the group consisting of a T helper epitope, an altered peptide ligand, and an adjuvant. In one embodiment, the immune stimulating agent is an adjuvant comprising carboxymethylcellulose, polyinosinic-polycytidylic acid, and poly-L-lysine double-stranded RNA (e.g., poly IC-LC t, e.g., hiltonol); an adjuvant comprising a water-and-oil emulsion (e.g., montanide); an adjuvant comprising a protein (e.g., a cytokine, a complement, GCSF, GM-CSF). In one embodiment, the immune modulating agent is an immune modulating agent described herein, e.g., a protein, e.g., an antibody which modulates the immune system (e.g., an anti-CTLA4 antibody, e.g., ipilimumab or tremelimumab, an anti-PD-1 antibody, an anti-PDL-1 antibody), a small molecule adjuvant (e.g., thalidomide or a thalidomide derivative, e.g., lenalidomide). In one embodiment, the kit further comprises instructions for administering an immune stimulating agent and/or immune modulating agent in combination with a peptide or peptides described herein or a composition described herein.

[0068] In one embodiment, the kit further comprises an additional immunogenic peptide, e.g., an immunogenic peptide from WO or a derivative thereof, e.g., an immunogenic WT1 peptide described herein. Other immunogenic peptides include, but are not limited to, an immunogenic peptide from MUC1, an immunogenic peptide from gp100, an immunogenic peptide from TRP-2, an immunogenic peptide from MAGI, an immunogenic peptide from NY-ESO1, an immunogenic peptide from HER-2; and an immunogenic peptide from AIM2. In one embodiment, the kit further comprises instructions for administering an additional immunogenic peptide, e.g., a WT1 peptide, in combination with a peptide or peptides described herein or a composition described herein.

[0069] In another aspect, the disclosure features an article of manufacture comprising: a container; and a composition contained within the container, wherein the composition is a composition described herein. The container can have a label indicating that the composition is for use in inducing an immune response in a mammal, e.g., a human. The label can further indicate that the composition is to be administered to a mammal having, suspected of having, or at risk of developing, cancer, e.g., a cancer described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma or a subject having a precancerous condition, e.g., smoldering multiple myeloma. The article of manufacture can also include instructions for administering the composition to the mammal, e.g., human. The composition can be, e.g., in solution, dried, or lyophilized.

[0070] In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck.

[0071] In yet another aspect, the disclosure features a method for inducing an immune response in a subject, which method includes the step of delivering, e.g., administering, to a subject one or more of any of the isolated peptides described herein and/or a composition described herein. In one embodiment, the subject is administered at least two, e.g., 2, 3 or 4, of the peptides from Group A. For example, the subject can be administered two or more of a non-spliced XBP1 peptide from Group A, a spliced XBP1 peptide from Group A, a CD138 peptide from Group A, a CS-1 peptide from Group A, and combinations thereof. In one embodiment, the subject is administered a non-spliced XBP1 peptide from Group A (e.g., a non-spliced XBP1 peptide comprising SEQ ID NO:6), a spliced XBP1 peptide from Group A (e.g., a spliced XBP1 peptide comprising SEQ ID NO:10), a CD138 peptide from Group A (e.g., a CD138 peptide comprising SEQ ID NO:12) and a CS-1 peptide from Group A (e.g., a CS-1 peptide comprising SEQ ID NO:16). In one embodiment, the subject is administered at least two, e.g., 2, 3 or 4, of the peptides from Group B. For example, the subject can be administered a non-spliced XBP1 peptide from Group B, a spliced XBP1 peptide from Group B, a CD138 peptide from Group B, a CS-1 peptide from Group B, and combinations thereof. In one embodiment, the subject is administered a non-spliced XBP1 peptide from Group B (e.g., a non-spliced XBP1 peptide comprising SEQ ID NO:29), a spliced XBP1 peptide from Group B (e.g., a spliced XBP1 peptide comprising SEQ ID NO:30), a CD138 peptide from Group B (e.g., a CD138 peptide comprising SEQ ID NO:31) and a CS-1 peptide from Group B (e.g., a CS-1 peptide comprising SEQ ID NO:32). In one embodiment, the subject is administered at least two (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more) of the peptides from Group C. In another embodiment, the subject is administered two or more peptides from a Group A and Group C or Group B and Group C.

[0072] The method can also include the step of, after delivering the one or more peptides or composition to the subject, determining if an immune response occurred in the subject. The one or more peptides can be delivered to the subject as a pharmaceutical composition, e.g., a pharmaceutical composition described herein. The subject can be, e.g., a mammal (e.g., a human) or any other subject described herein. The subject can have, be suspected of having, at risk of developing, or in remission from cancer, e.g., a cancer described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma. In one embodiment, the subject has a precancerous condition, e.g., smoldering multiple myeloma. In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck.

[0073] In some embodiments, the method can include determining whether the cancer cell (or cells) expresses one or more of XBP1, CD138, or CS-1.

[0074] In some embodiments, the method can further include administering to the subject one or more additional treatment, e.g., a chemotherapeutic agent, ionizing radiation, surgery or one or more additional immunotherapy agents. The one or more forms of ionizing radiation can be, e.g., gamma-irradiation, X-irradiation, or beta-irradiation. The one or more chemotherapeutic agents can be a chemotherapeutic agent described herein, e.g., a chemotherapeutic agent selected from the group consisting of a platinum based agent, a taxane, a topisomerase inhibitor, an antimetabolite, an alkylating agent, a protease inhibitor, and a vinca alkaloid. Exemplary chemotherapeutic agents, include, but are not limited to: cisplatin, carboplatin procarbazine, mechlorethamine, cyclophosphamide, camptothecin, adriamycin, ifosfamide, melphalan, chlorambucil, bisulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, mitomycin, etoposide, verampil, podophyllotoxin, taxol, transplatinum, 5-flurouracil, vincristin, vinblastin, methotrexate, and an analog of any of the aforementioned. The method can also include administering to the subject one or more immune stimulating agents, e.g., one or more immune stimulating agents described herein.

[0075] In one embodiment, the method further comprises administering an additional immunogenic peptide, e.g., an immunogenic peptide from WT1 or a derivative thereof, e.g., an immunogenic WT1 peptide described herein, in combination with the one or more peptides described herein. Other immunogenic peptides include, but are not limited to, an immunogenic peptide from MUC1, an immunogenic peptide from gp100, an immunogenic peptide from TRP-2, an immunogenic peptide from MAGI, an immunogenic peptide from NY-ESO1, an immunogenic peptide from HER-2; and an immunogenic peptide from AIM2.

[0076] In some embodiments, the delivering comprises administering to the subject the one or more peptides from Group A, Group B and/or Group C or a composition described herein. In some embodiments, the delivering comprises administering to the subject one or more nucleic acids, each of which comprises a nucleotide sequence encoding the one or more peptides, the nucleotide sequence being operably-linked to an expression control sequence. The nucleic acid can be in a recombinant cell transfected with the nucleic acid and expressing the one or more peptides. The recombinant cell can be a transfected cell, or the progeny of a transfected cell, made by transfecting a cell obtained from the subject. The recombinant cell can be an antigen presenting cell such as, but not limited to, a dendritic cell, a macrophage, a monocyte, or a B cell.

[0077] In some embodiments of any of the above-described methods, the delivering includes: contacting the one or more peptides to a cell; and after contacting the one or more peptides to the cell, delivering the cell to the subject. The cell can be, e.g., an antigen presenting cell such as any of those described herein. The cell can be, e.g., a cell, or the progeny of a cell, obtained from the subject. In some embodiments, the cell can be a cell, or the progeny of a cell, obtained from another subject of the same species as the subject. The other subject can express at least one MHC molecule in common with the subject. The at least one MHC molecule can be, e.g., an MHC class I molecule such as an HLA-A2 molecule and/or an HLA-A24 molecule.

[0078] In another aspect, the disclosure features a method for treating a subject having cancer, e.g., a cancer described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma or a subject having a precancerous condition, e.g., smoldering multiple myeloma. In one embodiment, the method includes the step of administering to a subject one or more (e.g., one, two, three, four, five, six, seven, eight, nine, 10, 11, 12, 13, 14, 15, 16, 17, or 18) of any of the peptides from Group A or a composition described herein, wherein the subject has, or is at risk of developing, a cancer, e.g., a cancer described herein, e.g., breast cancer (e.g., invasive lobular, invasive ductal, mixed lobular and ductal, intraductal cribriform, invasive lobular and ductal, invasive carcinoma), colon cancer (e.g., colon adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma or the subject has a precancerous condition, e.g., smoldering multiple myeloma. In one embodiment, the method includes the step of administering to a subject one or more (e.g., one, two, three, or four) of any of the peptides from Group B or a composition described herein, wherein the subject has, or is at risk of developing, a cancer, a cancer described herein, e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer (e.g., invasive lobular, invasive ductal, mixed lobular and ductal, intraductal cribriform, invasive lobular and ductal, invasive carcinoma), colon cancer (e.g., colon adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma. In one embodiment, the method includes the step of administering to a subject one or more (e.g., one, two, three, four, five, six, seven, eight, nine, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 20 or more) of any of the peptides from Group C or a composition described herein, wherein the subject has, or is at risk of developing, a cancer, e.g., a cancer described herein, e.g., breast cancer (e.g., invasive lobular, invasive ductal, mixed lobular and ductal, intraductal cribriform, invasive lobular and ductal, invasive carcinoma), colon cancer (e.g., colon adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma or the subject has a precancerous condition, e.g., smoldering multiple myeloma.

[0079] In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck.

[0080] In one embodiment, the subject is administered at least two, e.g., 2, 3 or 4, of the peptides from Group A. For example, the subject can be administered two or more of a non-spliced XBP1 peptide from Group A, a spliced XBP1 peptide from Group A, a CD138 peptide from Group A, a CS-1 peptide from Group A, and combinations thereof. In one embodiment, the subject is administered a non-spliced XBP1 peptide from Group A (e.g., a non-spliced XBP1 peptide comprising SEQ ID NO:6), a spliced XBP1 peptide from Group A (e.g., a spliced XBP1 peptide comprising SEQ ID NO:10), a CD138 peptide from Group A (e.g., a CD138 peptide comprising SEQ ID NO:12) and a CS-1 peptide (e.g., a CS-1 peptide comprising SEQ ID NO:16). The one or more peptides can be delivered to the subject as a pharmaceutical composition, e.g., a pharmaceutical composition from Group A.

[0081] In one embodiment, the subject is administered at least two, e.g., 2, 3 or 4, of the peptides from Group B. For example, the subject can be administered a non-spliced XBP1 peptide from Group B, a spliced XBP1 peptide from Group B, a CD138 peptide from Group B, a CS-1 peptide from Group B, and combinations thereof. In one embodiment, the subject is administered a non-spliced XBP1 peptide from Group B (e.g., a non-spliced XBP1 peptide comprising SEQ ID NO:29), a spliced XBP1 peptide from Group B (e.g., a spliced XBP1 peptide comprising SEQ ID NO:30), a CD138 peptide from Group B (e.g., a CD138 peptide comprising SEQ ID NO:31) and a CS-1 peptide from Group B (e.g., a CS-1 peptide comprising SEQ ID NO:32).

[0082] In one embodiment, the subject is administered at least two (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more) of the peptides from Group C. In another embodiment, the subject is administered two or more peptides from a Group A and Group C or Group B and Group C.

[0083] In one embodiment, the method further comprises administering an additional agent to the subject, e.g., administering a chemotherapeutic agent and/or an immune stimulating agent and/or an immune modulating agent. In one embodiment, the additional agent is an immune stimulating agent, e.g., an immune stimulating agent described herein. In one embodiment, the immune stimulating agent is an adjuvant comprising carboxymethylcellulose, polyinosinic-polycytidylic acid, and poly-L-lysine double-stranded RNA (e.g., poly IC-LC, e.g., hiltonol); an adjuvant comprising a water-and-oil emulsion (e.g., montanide); an adjuvant comprising a protein (e.g., a cytokine, a complement, GCSF, GM-CSF). In one embodiment, the additional agent is an immune modulating agent, e.g., an immune modulating agent described herein. In one embodiment, the immune modulating agent is a protein, e.g., an antibody which activates the immune system (e.g., an anti-CTLA4 antibody, e.g., ipilimumab or tremelimumab, an anti-PD-1 antibody, an anti-PDL-1 antibody); a small molecule adjuvant (e.g., thalidomide or a thalidomide derivative, e.g., lenalidomide. In one embodiment, the method comprises administering an additional immunogenic peptide, e.g., an immunogenic peptide from WT1 or a derivative thereof, e.g., a WT1 peptide described herein, in combination with the one or more of the peptides. Other immunogenic peptides include, but are not limited to, an immunogenic peptide from MUC1, an immunogenic peptide from gp100, an immunogenic peptide from TRP-2, an immunogenic peptide from MAGI, an immunogenic peptide from NY-ESO1, an immunogenic peptide from HER-2; and an immunogenic peptide from AIM2.

[0084] In one embodiment, the method further comprises administered one or more additional dose of a peptide from Group A. Group B and/or Group C or composition described herein. In one embodiment, the subject is administered one or more additional dose about 14 days after the previous dose, e.g., the subject is administered 2, 3, 4, 5, 6, 7, 8, 9 or 10 doses of a peptide from Group A, Group B and/or Group C or composition described herein, every other week.

[0085] In another aspect, the disclosure features a method for selecting a treatment for a mammal in need thereof. The method includes the steps of: determining if one or more cancer cells of a cancer in a mammal express XBP1, e.g., a cancer described herein, e.g., a breast cancer cell, a colon cancer cell, a pancreatic cancer cell, prostate cancer, a blood cell, e.g., a plasma cell; and if one or more of the cancer cells express XBP1, selecting as a therapeutic agent for the mammal one or more of the peptides from Group A, Group B and/or Group C, fusion proteins comprising such peptides, or compositions described herein. The method can also include the step of, after determining that one or more of the cells of the cancer express XBP1, delivering to the subject one or more of the peptides from Group A, Group B and/or Group C, fusion proteins comprising such peptides, or compositions described herein.

[0086] In yet another aspect, the disclosure features a method for selecting a treatment for a mammal with cancer. The method includes the steps of: determining if one or more cancer cells of a cancer in a mammal express CD138, e.g., a cancer described herein, e.g., a breast cancer cell, a colon cancer cell, a pancreatic cancer cell, a prostate cancer cell, a blood cell, e.g., a plasma cell; and if one or more of the cancer cells express CD138, selecting as a therapeutic agent for the mammal one or more of the peptides from Group A, Group B and/or Group C, fusion proteins comprising such peptides, or compositions described herein. The method can also include the step of, after determining that one or more of the cells of the cancer express CD138, delivering to the subject the one or more of the peptides from Group A, group B and/or Group C, fusion proteins comprising such peptides, or compositions described herein.

[0087] In another aspect, the disclosure features a method for selecting a treatment for a mammal in need thereof. The method includes determining if one or more cancer cells of a cancer in a mammal express CS-1, e.g., a cancer described herein, e.g., a breast cancer cell, a colon cancer cell, a pancreatic cancer cell, a prostate cancer cell, a blood cell, e.g., a plasma cell; and if one or more of the cancer cells express CS-1, selecting as a therapeutic agent for the mammal one or more of the peptides from Group A, Group B and/or Group C, fusion proteins comprising such peptides, or compositions described herein. The method can also include the step of, after determining that one or more of the cells of the cancer express CS1, delivering to the subject the selected one or more of the peptides from Group A, group B and/or Group C, fusion proteins comprising such peptides, or compositions described herein.

[0088] In yet another aspect, the disclosure features a method for selecting a therapeutic agent for a mammal having a cancer, e.g., a cancer described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, or a precancerous disorder, e.g., smoldering multiple myeloma. The method includes the step of, if one or more cancer cells of the mammal expresses XBP1, CD138 and/or CS-1, selecting as a therapeutic agent for the mammal one or more of the peptides from Group A, Group B and/or Group C, fusion proteins comprising such peptides, or compositions described herein. The method can also include the step of, after determining that one or more of the cells of the cancer express XBP1, CD138 and/or CS-1, delivering to the subject the selected one or more of the peptides from Group A, Group B and/or group C, fusion proteins comprising such peptides, or compositions described herein.

[0089] In some embodiments of any of the above methods, the subject or mammal can be one who has received a therapy for a cancer, e.g., a cancer described herein, e.g., breast, colon, pancreatic, prostate cancer, and was non-responsive to the therapy, e.g., the peptides from Group A, Group B and/or Group C, fusion proteins comprising such peptides, or compositions described herein may be a second-line, third line or fourth-line treatment.

[0090] In yet another aspect, the disclosure features a composition described herein and (ii) a major histocompatibility complex (MHC) molecule multimer, wherein the multimer comprises two or more (e.g., two, three, four, five, six, seven, eight, nine, or 10 or more) peptide-binding regions of an MHC molecule. In some embodiments, each peptide-binding region has a peptide from Group A, Group B, and/or Group C bound to it. In some embodiments, each peptide-binding region has a peptide from Group A, Group B and/or Group C non-covalently or covalently bound to it. The MHC molecule multimer can comprise two or more (e.g., two, three, four, five, six, seven, eight, nine, or 10 or more) entire MHC molecules. The MHC molecule multimer can comprise a human MHC molecule. The MHC molecule multimer can comprise an MHC class I molecule such as an HLA-A molecule, e.g., an HLA-A2 molecule or HLA-A24 molecule.

[0091] In some embodiments, the two or more peptide-binding regions can be from the same MHC molecule. In some embodiments, the two or more peptide-binding regions are from different MHC molecules. In some embodiments, the two or more peptide-binding regions can be a mixture of at least two (e.g., two, three, four, five, six, seven, eight, nine, or 10 or more) regions from the same MHC molecule and at least one (e.g., one, two, three, four, five, six, seven, eight, nine, or 10 or more) regions from a different MHC molecule.

[0092] In some embodiments, the MHC molecule multimer is capable of binding to at least one of the one of more peptides of the composition.

[0093] In some embodiments, the composition can be detectably labeled. For example, one or more of the peptides and/or one or more of the peptide-binding regions can be detectably labeled. In some embodiments, at least one of the one or more MHC molecule multimers or at least one of the one or more peptides are detectably labeled.

[0094] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.

[0095] Other features and advantages of the invention, e.g., methods for inducing an immune response in a subject, will be apparent from the following description, from the drawings and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

[0096] FIG. 1a is a bar graph showing the HLA-A2 binding capacity of a multipeptide cocktail. The Y-axis represents the mean fluorescence intensity, and the X-axis represents the peptide concentration of the cocktail. Influenza virus matrix protein.sub.58-66 (IVMP.sub.58-66;GILGFVFTL) (SEQ ID NO: 25) was used as an HLA-A2-specific positive control peptide.

[0097] FIG. 1b is a bar graph showing the HLA-A2 stability of a multipeptide cocktail using T2 cells. The Y-axis represents the mean fluorescence intensity, and the X-axis represents the time after Brefeldin A (BFA) treatment. Influenza virus matrix protein.sub.58-66 (WMP.sub.58-66;GILGFVFTL) (SEQ ID NO: 25) was used as an HLA-A2-specific positive control peptide.

[0098] FIG. 2 is a series of bar graphs showing the distinct phenotype of multipeptide-specific CTL (MP-CTL). The Y-axes indicate the percentage of cells in a given population.

[0099] FIG. 3 is a series of bar graphs showing IFN-.gamma. production by MP-CTL in response to HLA-A2.sup.+ MM cell lines. The Y-axes indicates the percentage of IFN-.gamma.+ cells in a given population.

[0100] FIG. 4 shows the induction of MP-CTL proliferation by stimulation with HLA-A2+MM cells, including primary myeloma cells and multiple myeloma cell lines. The upper panels are representative dot plots from flow cytometry analyses. The Y-axes indicate CD8 expression, and the X-axes indicate a decrease in CFSE staining which is a direct measure of cell proliferation. The lower panels are bar graphs showing the MP-CTL proliferative responses to primary multiple myeloma cells (lower left panel) and multiple myeloma cell lines (lower right panel). The Y-axes in the bar graphs indicate the percentage of proliferating MP-CTL, and the X-axes indicate the source of the stimulatory MM cells tested.

[0101] FIG. 5 is a series of graphs showing the cytotoxic activity of MP-CTL against HLA-A2+MM cells, including primary MM cells and cell lines. The Y-axes indicate the percent cytotoxicity, and the X-axes indicate the ratio of effector cells (MP-CTL) to target cells.

[0102] FIG. 6a is a series of dot plots showing the peptide-specific response of multipeptide-specific CTL generated from a single donor (Donor A). The Y-axes indicate CD107.alpha. expression levels, and the X-axes indicate IFN-.gamma. expression levels.

[0103] FIG. 6b is a series of bar graphs showing peptide-specific response of multipeptide-specific CTL generated from three donors (Donor B, Donor C and Donor D). The Y-axes indicate the percentage of CD107.alpha.+ cells (upper panels) or IFN-.gamma.+ cells (lower panels), and the X-axes indicate the peptide presented by K562-A2 cells.

[0104] FIG. 7 is a table showing the relative expression of unspliced XBP1 and spliced XBP1 in various cancer cell lines. Relative expression levels are indicated with plus or minus signs and also with numbers indicating number of plus signs.

[0105] FIG. 8a is a series of histograms showing the proliferation response at day 6 of XBP1-CTL to HLA-A2+ breast cancer cell lines. The X-axes indicate a decrease in CFSE staining, a direct measure of cell proliferation.

[0106] FIG. 8b is a series of histograms showing the proliferation response at day 7 of XBP1-CTL to HLA-A2+ breast cancer cells. The X-axes indicate a decrease in CFSE staining, a direct measure of cell proliferation FIG. 9 is a series of dot plots showing IFN-.gamma. production and cell activation (CD69 expression) of XBP1-CTL to HLA-A2+ breast cancer cell lines. The Y-axes indicate CD69 expression and the X-axes indicate IFN-.gamma. expression.

[0107] FIG. 10 is a series of dot plots showing the degranulation (CD107.alpha.) of XBP1-CTL to HLA-A2+ breast cancer cell lines. The Y-axes indicate CD107.alpha. expression, and the X-axes indicate CD8 expression.

[0108] FIG. 11 is a series of histograms showing the proliferation response of XBP1-CTL to HLA-A2+ pancreatic and colon cancer cell lines. The X-axes indicate a decrease in CFSE staining, a direct measure of cell proliferation.

[0109] FIG. 12 is a series of dot plots showing the IFN-.gamma. production and degranulation responses of XBP1-CTL to HLA-A2+ pancreatic and colon cancer cell lines. The Y-axes indicate CD107.alpha. expression, and the X-axes indicate IFN-.gamma. expression.

[0110] FIG. 13 is a table showing the relative CD138 expression in various cancer cell lines. Relative expression is indicated with plus or minus signs and also with numbers indicating number of plus signs.

[0111] FIG. 14 is a table showing the relative CS1 expression in various cancer cell lines. Relative expression is indicated with plus or minus signs and also with numbers indicating number of plus signs.

[0112] FIGS. 15a and b are a series of bar graphs showing the increase in CD8+ CTL induced by a cocktail of immunogenic XBP1-unspliced, XBP1-spliced, CD138, and CS-1 HLA-A2-specific peptides from T cells of different smoldering multiple myeloma patients. The Y-axes in the panels on the left indicate the percentage of CD3+CD8+ CTL, the Y-axes in the panels on the right indicate the percentage of CD4+Th cells, and the X-axes indicate the number of peptide stimulations prior to phenotypic analysis.

[0113] FIG. 16a is a series of histograms showing the proliferation response of MP-CTL generated from a smoldering multiple myeloma patient to myeloma cells in a HLA-A2-restricted manner. The X-axes indicate a decrease in CFSE staining, a direct measure of cell proliferation. The response 5 days after stimulation is shown in the upper panels, response after 6 days is shown in the middle panels, and the response after 7 days is shown in the lower panels.

[0114] FIG. 16b is a series of histograms showing the proliferation response of MP-CTL generated from a second smoldering multiple myeloma patient in response to myeloma cells in a HLA-A2-restricted manner. The Y-axes indicate numbers of cells, and the X-axes indicate CFSE staining. The response 5 days after stimulation is shown in the upper panels, response after 6 days is shown in the middle panels, and the response after 7 days is shown in the lower panels.

[0115] FIG. 17a is a series of dot plots showing the IFN-.gamma. production of MP-CTL generated from a smoldering multiple myeloma patient in response to myeloma cell lines in a HLA-A2-restricted manner. The Y-axes indicate IFN-.gamma. expression, and the X-axes indicate CD8 expression.

[0116] FIG. 17b is a series of bar graphs showing the IFN-.gamma. production of MP-CTL generated from four smoldering multiple myeloma patients in response to myeloma cell lines in a HLA-A2-restricted manner. The Y-axes indicate the percentage of IFN-.gamma.+ cells, and the X-axes indicate the type of cells used to stimulate the MP-CTL.

[0117] FIG. 18a is a series of dot plots showing the degranulation of MP-CTL generated from a smoldering multiple myeloma patient in response to myeloma cell lines in a HLA-A2-restricted manner. The Y-axes indicate CD107.alpha. expression, and the X-axes indicate CD8 expression.

[0118] FIG. 18b is a series of bar graphs showing the degranulation of MP-CTL generated from four smoldering multiple myeloma patients in response to myeloma cell lines in a HLA-A2-restricted manner. The Y-axes indicate the percentage of CD107.alpha.+ cells, and the X-axes indicate the type of cells used to stimulate the MP-CTL.

[0119] FIG. 19a is a series of dot plots showing polyfunctional IFN-.gamma. production and degranulation (CD107.alpha.) among CD3+CD8+CD137+MP-CTL generated from a smoldering multiple myeloma patient in response to K562-A2 cells presenting an individual peptide. The Y-axes indicates CD107.alpha. expression and the Y-axes indicates + cells and the X-axes indicate IFN-.gamma. expression.

[0120] FIG. 19b is a series of bar graphs showing polyfunctional IFN-.gamma. production and degranulation (CD107.alpha.) among CD3+CD8+CD137+MP-CTL generated from three smoldering multiple myeloma patients in response to K562-A2 cells presenting an individual peptide. The Y-axes indicate the percentage of CD3+CD8+CD137+ cells that express both IFN-.gamma. and CD107.alpha., and the X-axes indicate the peptide presented by the K562 A2+ cells.

[0121] FIG. 19c is a summary bar graph showing both IFN-.gamma. production and degranulation (CD107.alpha.) among CD3+CD8+CD137+MP-CTL generated from three smoldering multiple myeloma patients in response to K562-A2 cells presenting an individual peptide. The Y-axes indicate the percentage of CD3+CD8+CD137+ cells that express both IFN-.gamma. and CD107.alpha. and the X-axes indicate the peptide presented by the K562-A2 cells.

[0122] FIG. 19d is a summary bar graph from two separate experiments showing both IFN-.gamma. production and degranulation (CD107.alpha.) among CD3+CD8+CD137+MP-CTL generated from three smoldering multiple myeloma patients in response to K562-A2 cells presenting an individual peptide. The Y-axes indicate the percentage of CD3+CD8+CD137+ cells that express both IFN-.gamma. and CD107.alpha. and the X-axes indicate the peptide presented by the K562-A2 cells.

[0123] FIG. 20a is a series of dot plots (upper panels) and bar graphs (lower panels) showing an increase of memory CD8.sup.+ T cells in MP-CTL generated from four smoldering multiple myeloma patients. The Y-axes in the dot plots show CCR7 expression, and the X-axes indicate CD45RP expression. The Y-axes in the bar graphs indicate the percentage of cells positive for the T cell subsets defined on the, X-axes.

[0124] FIG. 20b is a series of bar graphs showing the increase in effector memory (EM) cells among MP-CTL generated from two smoldering multiple myeloma patients. Increasing the number of peptide stimulations from 4 to 7 resulted in an increase in the percentage of EM type cells. The Y-axes in the bar graphs indicate the percentage of cells positive for the T cell subsets defined on the X-axes.

[0125] FIG. 20c is a series of histograms showing the increase in IFN-.gamma.+CD107.alpha.+ among effector memory cells (EM) and terminal effector cells (TE) from three smoldering multiple myeloma patients. The Y-axes indicate the percentage of double positive IFN-.gamma.+CD107.alpha.+ cells, and the X-axes indicate the type of cells used to stimulate the MP-CTL.

[0126] FIG. 20d is a series of bar graphs showing CD69 activation for the naive and memory CD8+ T cell subsets in MP-CTL generated from four smoldering multiple myeloma patients in response to MM cell lines (HLA-A2+U266 cells or HLA-A2- RPMI cells). The Y-axes show the percentage of CD69+ cell, and the X-axes indicate the CTL subpopulations.

[0127] FIG. 21 is a graph showing the affinity of peptides from unspliced XBP1, spliced XBP1, CD138, and CS1 for HLA-A24. T2 cells were exposed to indicated peptides at a concentration of 1 mg/ml. HIV envelope protein.sub.583-591 (RYLKDQQLL; SEQ ID NO: 537) was used as an HLA-A24-specific positive control peptide.

[0128] FIG. 22 is a graph showing the affinity of unspliced XBP1 peptide 4 (SEQ ID NO: 35), unspliced XBP1 peptide 7 (SEQ ID NO: 29), and spliced peptide 1 (SEQ ID NO: 30) for HLA-A24. T2 cells were exposed to peptides at concentrations indicated.

[0129] FIG. 23 is a graph showing the affinity of CD138 peptides 1 (SEQ ID NO: 31), 3 (SEQ ID NO: 41), and 4 (SEQ ID NO: 42) for HLA-A24. T2 cells were exposed to peptides at concentrations indicated.

[0130] FIG. 24 is a graph showing the affinity of CS1 peptides 3 (SEQ ID NO: 48) and 5 (SEQ ID NO: 32) for HLA-A24. T2 cells were exposed to peptides at concentrations indicated.

[0131] FIG. 25 is a schematic representation of the method used to generate peptide-specific CTL. APCs presenting indicated peptides were used to stimulate CD3+T lymphocytes from donors to generate peptide-specific CTL.

[0132] FIG. 26a is a bar graph showing the increase in CD8+ T cells induced by unspliced XBP1 peptide 7 presented on T lymphocytes from two donors (Donor A and Donor B). The Y-axes indicate the percentage of CD8+T lymphocytes, and the X-axes indicate the number of peptide stimulations prior to phenotypic analysis.

[0133] FIG. 26b is a bar graph showing the increase in CD8+ T cells induced by spliced XBP1 peptide 1 presented on T lymphocytes from two donors (Donor A and Donor B). The Y-axes indicate the percentage of CD8+T lymphocytes, and the X-axes indicate the number of peptide stimulations prior to phenotypic analysis.

[0134] FIG. 26c is a bar graph showing the increase in CD8+ T cells induced by CD138 peptide 1 presented on T lymphocytes from two donors (Donor A and Donor B). The Y-axes indicate the percentage of CD8+T lymphocytes, and the X-axes indicate the number of peptide stimulations prior to phenotypic analysis.

[0135] FIG. 26d is a bar graph showing the increase in CD8+ T cells induced by CS1 peptide 1 presented on T lymphocytes from two donors (Donor A and Donor B). The Y-axes indicate the percentage of CD8+T lymphocytes, and the X-axes indicate the number of peptide stimulations prior to phenotypic analysis.

[0136] FIG. 27 is a schematic representation of the method used to evaluate the response of peptide-specific CTL to various multiple myeloma tumor cells. Peptide-specific CTL were incubated for 5 hours with KMS, OPM1, or U266 multiple myeloma cells and assayed for IFN-.gamma. production, CD107.alpha. upregulation, CD8 T cell proliferation, or IL-2 production.

[0137] FIG. 28 is a series of dot plots showing IFN-.gamma. production and CD8 expression of peptide-specific CTL in response to multiple myeloma cells. The Y-axes indicate the level of IFN-.gamma. expression, and the X-axes indicate CD8 expression. The peptide specificity of the CTL population analyzed is shown on the left. CD8+, IFN-.gamma.+ populations are represented in boxed regions.

[0138] FIG. 29a is a series of dot plots showing IFN-.gamma. expression in populations and subpopulations of CTL. CTL specific for unspliced XBP1 peptide 7 (SEQ ID NO: 29) were stimulated with KMS11 cells. Populations represented in boxed regions are indicated on graphs.

[0139] FIG. 29b is a series of dot plots showing IFN-.gamma. expression in populations and subpopulations of CTL. CTL specific for spliced XBP1 peptide 1 (SEQ ID NO: 30) were stimulated with KMS11 cells. Populations represented in boxed regions are indicated on graphs.

[0140] FIG. 29c is a series of dot plots showing IFN-.gamma. expression in populations and subpopulations of CTL. CTL specific for CD138 peptide 1 (SEQ ID NO: 31) were stimulated with KMS11 cells. Populations represented in boxed regions are indicated on graphs.

[0141] FIG. 29d is a series of dot plots showing IFN-.gamma. expression in populations and subpopulations of CTL. CTL specific for CS1 peptide 5 (SEQ ID NO: 32) were stimulated with KMS11 cells. Populations represented in boxed regions are indicated on graphs.

[0142] FIG. 30 is a schematic representation of the CD107.alpha. detection assay used to measure degranulation. Release of lytic granules due to degranulation of tumor cells causes upregulation of CD107.alpha. in CTL, which can be detected by anti-CD107.alpha. antibodies.

[0143] FIG. 31 is a series of dot plots showing IFN-.gamma. expression and degranulation in peptide-specific CTL. CTL were either unstimulated or stimulated with KMS11 or OPM1 cells, as indicated at top. Peptide specificities of CTL are shown on the left. Y-axes represent CD107.alpha. expression, and X-axes represent IFN-.gamma. expression.

[0144] FIG. 32 is a schematic representation of the assay used to measure proliferation of peptide-specific CTL in response to multiple myeloma cells. Peptide-specific CTL and irradiated multiple myeloma cells were incubated together for 6 or 8 days, and proliferation of CTL was measured by incorporation of CFSE.

[0145] FIG. 33a is a series of histograms showing the proliferation response at day 6 of peptide-specific CTL to myeloma cells. Peptide specificities of CTL are shown on the left. The X-axes indicate a decrease in CFSE staining, a direct measure of cell proliferation.

[0146] FIG. 33b is a series of histograms showing the proliferation response at day 8 of peptide-specific CTL to myeloma cells. Peptide specificities of CTL are shown on the left. The X-axes indicate a decrease in CFSE staining, a direct measure of cell proliferation.

[0147] FIG. 34 is a series of dot plots showing IL-2 production of peptide-specific CTL in response to myeloma cells. Peptide specificities of CTL are shown on the left. Y-axes indicate IL-2 expression, and X-axes indicate CD8 expression. Boxed regions represent IL-2+CD8+ populations.

[0148] FIG. 35 is a series of dot plots showing IFN-.gamma. expression of peptide-specific CTL from Donor A in response to various colon cancer cell lines. CTL were either unstimulated or stimulated with SW80, WiDr, or LS180 cells, as indicated at top. Peptide specificities of CTL are shown on the left. Y-axes indicate IFN-.gamma. expression, and X-axes indicate CD8 expression. Boxed regions represent IFN-.gamma.+CD8+ populations.

[0149] FIG. 36 is a series of dot plots showing IFN-.gamma. expression and degranulation in peptide-specific CTL from Donor A in response to various colon cancer cell lines. CTL were either unstimulated or stimulated with SW80, WiDr, or LS180 cells, as indicated at top. Peptide specificities of CTL are shown on the left. Y-axes represent CD107.alpha. expression, and X-axes represent IFN-.gamma. expression.

[0150] FIG. 37 is a series of dot plots showing IFN-.gamma. expression of peptide-specific CTL from Donor B in response to various colon cancer cell lines. CTL were either unstimulated or stimulated with SW80, WiDr, or LS180 cells, as indicated at top. Peptide specificities of CTL are shown on the left. Y-axes indicate IFN-.gamma. expression, and X-axes indicate CD8 expression. Boxed regions represent IFN-.gamma.+CD8+ populations.

[0151] FIG. 38 is a series of dot plots showing IFN-.gamma. expression and degranulation in peptide-specific CTL from Donor B. CTL were either unstimulated or stimulated with SW80, WiDr, or LS180 cells, as indicated at top. Peptide specificities of CTL are shown on the left. Y-axes represent CD107.alpha. expression, and X-axes represent IFN-.gamma. expression.

[0152] FIG. 39a is a series of histograms showing the degranulation response of CD138 peptide-specific CTL to SW480 tumor cells. CTL and SW480 tumor cells were co-incubated at various cell:cell ratios, as indicated on the left, and analyzed for expression of CD107.alpha., IFN-.gamma., and IL-2, as indicated at top.

[0153] FIG. 39b is a series of histograms showing various response of CD138 peptide-specific CTL to LS180 tumor cells. CTL and LS180 tumor cells were co-incubated at various cell:cell ratios, as indicated on the left, and analyzed for expression of CD107.alpha., IFN-.gamma., and IL-2, as indicated at top.

[0154] FIG. 40a is a series of dot plots showing IFN-.gamma. expression of peptide-specific CTL from Donor A in response to different cancer cell types. CTL were incubated alone or co-incubated with either SW480 colon cancer cells or KMS11 multiple myeloma cells and analyzed for IFN-.gamma. expression. Peptide specificities of CTL are shown on the left. Y-axes indicate IFN-.gamma. expression, and X-axes indicate CD8 expression. Boxed regions represent IFN-.gamma.+CD8+ populations.

[0155] FIG. 40b is a series of dot plots showing degranulation and IFN-.gamma. expression of peptide-specific CTL from Donor A in response to different cancer cell types. CTL were incubated alone or co-incubated with either SW480 colon cancer cells or KMS11 multiple myeloma cells and analyzed for IFN-.gamma. expression. Peptide specificities of CTL are shown on the left. Y-axes indicate CD107.alpha. expression, and X-axes indicate IFN-.gamma. expression.

[0156] FIG. 41a is a series of dot plots showing IFN-.gamma. expression of peptide-specific CTL from Donor B in response to different cancer cell types. CTL were incubated alone or co-incubated with either SW480 colon cancer cells or KMS11 multiple myeloma cells and analyzed for IFN-.gamma. expression. Peptide specificities of CTL are shown on the left. Y-axes indicate IFN-.gamma. expression, and X-axes indicate CD8 expression. Boxed regions represent IFN-.gamma.+CD8+ populations.

[0157] FIG. 41b is a series of dot plots showing degranulation and IFN-.gamma. expression of peptide-specific CTL from Donor B in response to different cancer cell types. CTL were incubated alone or co-incubated with either SW480 colon cancer cells or KMS11 multiple myeloma cells and analyzed for IFN-.gamma. expression. Peptide specificities of CTL are shown on the left. Y-axes indicate CD107.alpha. expression, and X-axes indicate IFN-.gamma. expression.

[0158] FIG. 42 is a series of dot plots showing IFN-.gamma. expression of peptide-specific CTL from Donor B in response to various pancreatic cancer cell lines. CTL were incubated alone or co-incubated with 8902, PL45, or MiaPaca cells and analyzed for IFN-.gamma. expression. Peptide specificities of CTL are shown on the left. Y-axes indicate IFN-.gamma. expression, and X-axes indicate CD8 expression. Boxed regions represent IFN-.gamma.+CD8+ populations.

[0159] FIG. 43 is a series of histograms and dot plots showing various response of CD138 peptide-specific CTL to Panc1 pancreatic tumor cells. CTL and Panc1 cells were co-incubated at various cell:cell ratios of 1:1 or 1:5, as indicated on the left, and analyzed for expression of CD107.alpha., IFN-.gamma., and IL-2, as indicated at top. Y-axes of dot plots indicate CD107.alpha. expression, and X-axes of histograms and dot plots indicate IFN-.gamma. or IL-2 expression, as indicated below each graph.

[0160] FIG. 44 is a series of dot plots showing the phenotypic changes of T cells (naive T cells, central memory (CM), effector cells and effector memory (EM) CD8.sup.+ T cells) in CTL induced with a cocktail of heteroclitic unspliced XBP1 and heteroclitic spliced XBP1 peptides. The Y-axes in the dot plots show CCR7 expression, and the X-axes indicate CD45RP expression.

[0161] FIG. 45 is a series of bar graphs depicting the generation of central memory CD3+CD8+ T cells in three donors in response to a cocktail of heteroclitic unspliced XBP1 and heteroclitic spliced XBP1 peptides.

[0162] FIG. 46 is a series of bar graphs depicting the generation of effector memory CD3+CD8+ T cells in three donors in response to a cocktail of heteroclitic unspliced XBP1 and heteroclitic spliced XBP1 peptides.

[0163] FIG. 47 is a series of histograms showing the proliferation response of the XBP1 peptide cocktail-specific CTL to MB231 breast cancer cells. The response is broken into proliferation in CD45RO- non-memory cells and CD45RO+ memory cells, and within the memory cells which proportion are CD45RO+, CCR7+ central memory T cells and which are CD45RO+, CCR7- effector memory T cells.

[0164] FIG. 48 is a series of histograms showing the proliferation response of the XBP1 peptide cocktail-specific CTL to LS180 colon cancer cells. The response is broken into proliferation in CD45RO- non-memory cells and CD45RO+ memory cells, and within the memory cells which proportion are CD45RO+, CCR7+ central memory T cells and which are CD45RO+, CCR7- effector memory T cells.

[0165] FIG. 49 is a series of histograms showing the proliferation response of the XBP1 peptide cocktail-specific CTL to Panc1 pancreatic cancer cells. The response is broken into proliferation in CD45RO- non-memory cells and CD45RO+ memory cells, and within the memory cells which proportion are CD45RO+, CCR7+ central memory T cells and which are CD45RO+, CCR7- effector memory T cells.

[0166] FIG. 50 is a series of histograms showing various response of the XBP1 peptide cocktail-specific CTL to various tumor cells (MB231, MCF7, LS180, SW480, Panc1 and PL45). Central memory T-cells and effector memory T cells were analyzed for expression of IFN-.gamma..

[0167] FIG. 51 are bar graphs showing expression of IFN-.gamma. by effector memory T cells and central memory T cells of XBP1 cocktail specific-CTL to various tumor cells (MB231, MC7, LS180, SW480, Panc1 and PL45).

[0168] FIG. 52 is a series of histograms showing various response of the XBP1 peptide cocktail-specific CTL to various tumor cells (MB231, MCF7, LS180, SW480, Panc1 and PL45). Central memory T-cells and effector memory T cells were analyzed for expression of IL-2.

[0169] FIG. 53 depicts bar grafts showing expression of IL-2 by effector memory T cells and central memory T cells of XBP1 cocktail specific-CTL to various tumor cells (MB231, MC7, LS180, SW480, Panc1 and PL45).

[0170] FIG. 54 is a series of histograms showing various response of the XBP1 peptide cocktail-specific CTL to various tumor cells (MB231, MCF7, LS180, SW480, Panc1 and PL45). Central memory T-cells and effector memory T cells were analyzed for cytotoxicity.

[0171] FIG. 55 depicts bar graphs showing cytotoxicity of effector memory T cells and central memory T cells of XBP1 cocktail specific-CTL to various tumor cells (MB231, MC7, LS180, SW480, Panc1 and PL45).

[0172] FIG. 56 depicts bar graphs showing expression of Tbet by non-memory and memory T cells of XBP1 cocktail specific-CTL.

[0173] FIG. 57 depicts bar graphs showing expression of Tbet by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL.

[0174] FIG. 58 depicts a bar graphs showing expression of Tbet and IFN-.gamma. by non-memory T cells and memory T cells of XBP1 cocktail specific-CTL to various tumor cells (MB231, SW480 and Panc1).

[0175] FIG. 59 depicts bar graphs showing expression of Tbet and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to MB231 breast cancer cells.

[0176] FIG. 60 depicts bar graphs showing expression of Tbet and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to Panc1 pancreatic cancer cells.

[0177] FIG. 61 depicts bar graphs showing expression of Tbet and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to SW480 colon cancer cells.

[0178] FIG. 62 depicts bar graphs showing expression of Eomes by non-memory and memory T cells of XBP1 cocktail specific-CTL.

[0179] FIG. 63 depicts bar graphs showing expression of Eomes by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL.

[0180] FIG. 64 depicts a bar graph showing expression of Eomes and IFN-.gamma. by non-memory T cells and memory T cells of XBP1 cocktail specific-CTL to various tumor cells (MB231, SW480 and Panc1).

[0181] FIG. 65 depicts bar graphs showing expression of Eomes and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to MB231 breast cancer cells.

[0182] FIG. 66 depicts bar graphs showing expression of Eomes and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to Panc1 pancreatic cancer cells.

[0183] FIG. 67 depicts bar graphs showing expression of Eomes and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to SW480 colon cancer cells.

[0184] FIG. 68 depicts a bar graph showing expression of granzyme B by non-memory T cells and memory T cells of XBP1 cocktail specific-CTL to various tumor cells (MB231, SW480 and Panc1).

[0185] FIG. 69 depicts bar graphs showing expression of granzyme B and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to MB231 breast cancer cells.

[0186] FIG. 70 depicts bar graphs showing expression of granzyme B and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to Panc1 pancreatic cancer cells.

[0187] FIG. 71 depicts bar graphs showing expression of granzyme B and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to SW480 colon cancer cells.

[0188] FIG. 72 depicts a bar graph showing the number of non-memory T cells and memory T cells of XBP1 cocktail specific-CTL in the presence and absence of adjuvant, lenalidomide.

[0189] FIG. 73 depicts a bar graph showing the number of central memory T cells and effector memory T cells of XBP1 cocktail specific-CTL in the presence and absence of adjuvant, lenalidomide.

[0190] FIG. 74 depicts bar graphs showing the expression of CD40L, CD69 and CD38 in XBP1 cocktail specific-CTL in the presence and absence of adjuvant, lenalidomide.

[0191] FIG. 75 depicts bar graphs showing expression of Tbet and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to MB231 breast cancer cells in the presence or absence of adjuvant, lenalidomide.

[0192] FIG. 76 depicts bar graphs showing expression of Eomes and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to MB231 breast cancer cells in the presence or absence of adjuvant, lenalidomide.

[0193] FIG. 77 depicts bar graphs showing expression of Tbet and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to Panc1 pancreatic cancer cells in the presence or absence of adjuvant, lenalidomide.

[0194] FIG. 78 depicts bar graphs showing expression of Eomes and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to Panc1 pancreatic cancer cells in the presence or absence of adjuvant, lenalidomide.

[0195] FIG. 79 depicts bar graphs showing expression of Tbet and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to SW480 colon cancer cells in the presence or absence of adjuvant, lenalidomide.

[0196] FIG. 80 depicts bar graphs showing expression of Eomes and IFN-.gamma. by naive T-cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail specific-CTL to SW480 colon cancer cells in the presence or absence of adjuvant, lenalidomide.

[0197] FIG. 81 depicts bar graphs showing expression of granzyme and IFN-.gamma. by non-memory and memory T cells of XBP1 cocktail specific-CTL to MB231 breast cancer cells in the presence or absence of adjuvant, lenalidomide.

[0198] FIG. 82 depicts bar graphs showing expression of granzyme and IFN-.gamma. by non-memory and memory T cells of XBP1 cocktail specific-CTL to Panc1 pancreatic cancer cells in the presence or absence of adjuvant, lenalidomide.

[0199] FIG. 83 depicts bar graphs showing expression of granzyme and IFN-.gamma. by non-memory and memory T cells of XBP1 cocktail specific-CTL to SW480 colon cancer cells in the presence or absence of adjuvant, lenalidomide.

DETAILED DESCRIPTION

[0200] The disclosure features immunogenic XBP1-, CD138-, and CS-1-derived peptides (and pharmaceutical compositions thereof), which can be used to, e.g., induce an immune response (e.g., stimulate a CTL response), or stimulate the production of an antibody, in a subject. The peptides can be used in a variety of applications such as methods for inducing an immune response, methods for producing an antibody, and methods for treating a cancer (e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia (e.g., AML or CML) and a plasma cell disorder such as multiple myeloma or Waldenstrom's macroglobulinemia) or a precancerous disorder (e.g., smoldering multiple myeloma)). The peptides can also be included in MHC molecule multimer compositions and used in, e.g., methods for detecting a T cell in a population of cells.

[0201] A detailed description of the peptides as well as exemplary methods for making and using the peptides are set forth below.

[0202] Peptides

[0203] Group A Peptides.

[0204] The disclosure features isolated peptides ("Group A peptides") comprising an amino acid sequence that has sufficient identity with or is identical to any one of SEQ ID NOS:1-18 as depicted in Table 1.

TABLE-US-00001 TABLE 1 Examples of Group A peptides Amino Acid Amino Acid SEQ Protein of Origin Position Sequence ID NO: non-spliced XBP1 118-126 LLREKTHGL 1 non-spliced XBP1 185-193 NISPWILAV 2 non-spliced XBP1 190-198 ILAVLTLQI 3 non-spliced XBP1 193-201 VLTLQIQSL 4 non-spliced XBP1 111-119 KLLLENQLL 5 non-spliced XBP1 185-193 YISPWILAV 6 spliced XBP1 197-205 GILDNLDPV 7 spliced XBP1 194-202 ILLGILDNL 8 spliced XBP1 368-376 ELFPQLISV 9 spliced XBP1 368-376 YLFPQLISV 10 CD138 256-264 VIAGGLVGL 11 CD138 260-268 GLVGLIFAV 12 CD138 5-13 ALWLWLCAL 13 CD138 7-15 WLWLCALAL 14 CS1 236-245 LLLSLFVLGL 15 CS1 239-247 SLFVLGLFL 16 CS1 232-240 LLVPLLLSL 17 CS1 9-17 TLIYILWQL 18 Bolded residues indicate amino acid changes from the corresponding wild-type amino acid sequence.

[0205] Preferably, the isolated peptide from Group A is at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or 35 amino acids in length (e.g., between 9 and 35 amino acids in length, e.g., 9-30, 9-25, 9-20, 9-15 amino acids in length) and comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or is identical to an amino acid sequence of SEQ ID NOS:1-18. Other preferred peptides can be at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or 35 amino acids in length (e.g., between 9 and 35 amino acids in length, e.g., 9-30, 9-25, 9-20, 9-15 amino acids in length) and comprise an amino acid sequence of SEQ ID NOS:1-18, or an amino acid sequence with one, two, three or four substitutions of the amino acid sequence of SEQ ID NOs:1-18 The substitution can be a conservative or nonconservative substitution.

[0206] "Non-spliced XBP1" peptides from Group A include those peptides depicted in Table 1 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) consecutive amino acids from the non-spliced form of human XBP1 protein having 261 amino acids and the following sequence: MVVVAAAPNPADGTPKVLLLSGQPASAAGAPAGQALPLMVPAQRGASPEAASGGLPQ ARKRQRLTHLSPEEKALRRKLKNRVAAQTARDRKKARMSELEQQVVDLEEENQKLLLE NQLLREKTHGLVVENQELRQRLGMDALVAEEEAEAKGNEVRPVAGSAESAALRLRAPL QQVQAQLSPLQNISPWILAVLTLQIQSLISCWAFWTTWTQSCSSNALPQSLPAWRSSQRS TQKDPVPYQPPFLCQWGRHQPSWKPLMN (SEQ ID NO:19; Genbank Accession No.

[0207] NP_005071), and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:19. The amino acid positions referred to in Table 1 are based on SEQ ID NO: 19.

[0208] "Spliced XBP1" peptides from Group A include those peptides depicted in Table 1 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 31, 32, 33, 34 or 35) consecutive amino acids from the spliced form of human XBP1 (XBP1 spliced) protein having 376 amino acids and the following sequence: MVVVAAAPNPADGTPKVLLLSGQPASAAGAPAGQALPLMVPAQRGASPEAASGGLPQ ARKRQRLTHLSPEEKALRRKLKNRVAAQTARDRKKARMSELEQQVVDLEEENQKLLLE NQLLREKTHGLVVENQELRQRLGMDALVAEEEAEAKGNEVRPVAGSAESAAGAGPVV TPPEHLPMDSGGIDSSDSESDILLGILDNLDPVMFFKCPSPEPASLEELPEVYPEGPSSLPA SLSLSVGTSSAKLEAINELIRFDHIYTKPLVLEIPSETESQANVVVKIEEAPLSPSENDHPEF IVSVKEEPVEDDLVPELGISNLLSSSHCPKPSSCLLDAYSDCGYGGSLSPFSDMSSLLGVN HSWEDTFANELFPQLISV (SEQ ID NO: 20; Genbank Accession No. NP_001073007), and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:20. The amino acid positions referred to in Table 1 are based on SEQ ID NO: 20.

[0209] "CD138" peptides from Group A include those peptides depicted in Table 1 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) consecutive amino acids from the human CD138 protein having 310 amino acids and the following sequence: MRRAALWLWLCALALSLQPALPQIVATNLPPEDQDGSGDDSDNFSGSGAGALQDITLS QQTPSTWKDTQLLTAIPTSPEPTGLEATAAS TSTLPAGEGPKEGEAVVLPEVEPGLTARE QEATPRPRETTQLPTTHQASTTTATTAQEPATSHPHRDMQPGHHETSTPAGPS QADLHTP HTEDGGPSATERAAEDGASSQLPAAEGSGEQDFTFETSGENTAVVAVEPDRRNQSPVDQ GATGASQGLLDRKEVLGGVIAGGLVGLIFAVCLVGFMLYRMKKKDEGSYSLEEPKQAN GGAYQKPTKQEEFYA (SEQ ID NO:21; Genbank Accession No. NP_002988) and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:21. The amino acid positions referred to in Table 1 are based on SEQ ID NO: 21.

[0210] "CS-1" peptides from Group A include those peptides depicted in Table 1 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) consecutive amino acids from the human CS-1 protein having 335 amino acids and the following sequence: MAGSPTCLTLIYILWQLTGSAASGPVKELVGSVGGAVTFPLKSKVKQVDSIVWTFNTTP LVTIQPEGGTIIVTQNRNRERVDFPDGGYSLKLSKLKKNDSGIYYVGIYSSSLQQPSTQEY VLHVYEHLSKPKVTMGLQSNKNGTCVTNLTCCMEHGEEDVIYTWKALGQAANESHNG SILPISWRWGESDMTFICVARNPVSRNFSSPILARKLCEGAADDPDSSMVLLCLLLVPLLL SLFVLGLFLWFLKRERQEEYIEEKKRVDICRETPNICPHS GENTEYDTIPHTNRTILKEDPA NTVYSTVEIPKKMENPHSLLTMPDTPRLFAYENVI (SEQ ID NO:22; Genbank Accession No. NP_067004) and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:22. The amino acid positions referred to in Table 1 are based on SEQ ID NO: 22.

[0211] Group B Peptides.

[0212] The disclosure also features isolated peptides ("Group B peptides") comprising an amino acid sequence that has sufficient identity with or is identical to any one of SEQ ID NOS:29-50 as depicted in Table 2.

TABLE-US-00002 TABLE 2 Examples of Group B peptides Amino Acid Amino Acid SEQ Protein of Origin Position Sequence ID NO: non-spliced XBP1 188-196 PWILAVLTL 33 non-spliced XBP1 111-119 KLLLENQLL 5 non-spliced XBP1 129-137 ENQELRQRL 34 non-spliced XBP1 12-20 DGTPKVLLL 35 non-spliced XBP1 104-112 DLEEENQKL 36 non-spliced XBP1 216-224 SSNALPQSL 37 non-spliced XBP1 186-194 ISPWILAVL 29 spliced XBP1 224-232 VYPEGPSSL 30 spliced XBP1 340-348 GYGGSLSPF 38 spliced XBP1 347-355 PFSDMSSLL 39 spliced XBP1 111-119 KLLLENQLL 5 CD138 265-273 IFAVCLVGF 31 CD138 106-114 VLPEVEPGL 40 CD138 21-29 LPQIVATNL 41 CD138 13-21 LALSLQPAL 42 CD138 245-253 GLLDRKEVL 43 CD138 262-270 VGLIFAVCL 44 CD138 16-24 SLQPALPQI 45 CS1 87-95 GYSLKLSKL 46 CS1 82-90 DFPDGGYSL 47 CS1 321-329 TMPDTPRLF 48 CS1 185-193 RWGESDMTF 49 CS1 240-248 LFVLGLFLW 32 CS1 312-320 KMENPHSLL 50

[0213] Preferably, the isolated peptide from Group B is at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or 35 amino acids in length (e.g., between 9 and 35 amino acids in length, e.g., 9-30, 9-25, 9-20, 9-15 amino acids in length) and comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or is identical to an amino acid sequence of SEQ ID NOS:29-50. Other preferred peptides can be at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or 35 amino acids in length (e.g., between 9 and 35 amino acids in length, e.g., 9-30, 9-25, 9-20, 9-15 amino acids in length) and comprise an amino acid sequence of SEQ ID NOS:29-50, or an amino acid sequence with one, two, three or four substitutions of the amino acid sequence of SEQ ID NOs: 29-50. The substitution can be a conservative or nonconservative substitution.

[0214] "Non-spliced XBP1" peptides from Group B include those peptides depicted in Table 2 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, or 12) consecutive amino acids from the non-spliced form of human XBP1 protein having 261 amino acids and the amino acid sequence of SEQ ID NO:19, and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:19. Non-spliced XBP1 peptides from Group B include peptides having an amino acid sequence from SEQ ID NO: 19 that comprises part or all of any one of SEQ ID NOS: 29 and 33-37, e.g., a sequence that comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids N-terminal and/or C-terminal to any one of SEQ ID NOS: 29 and 33-37. The amino acid positions referred to in Table 2 are based SEQ ID NO: 19.

[0215] "Spliced XBP1" peptides from Group B include those peptides depicted in Table 2 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 31, 32, 33, 34 or 35) consecutive amino acids from the spliced form of human XBP1 (XBP1 spliced) protein having 376 amino acids and amino acid sequence of SEQ ID NO: 20, and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:20. Spliced XBP1 peptides from Group B include peptides having an amino acid sequence from SEQ ID NO: 20 that comprises part or all of any one of SEQ ID NOS: 30, 38 and 39, e.g., a sequence that comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids N-terminal and/or C-terminal to any one of SEQ ID NOS: 30, 38, and 39. The amino acid positions referred to in Table 2 are based on SEQ ID NO: 20.

[0216] "CD138" peptides from Group B include those peptides depicted in Table 2 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) consecutive amino acids from the human CD138 protein having 310 amino acids and the amino acid sequence of SEQ ID NO:21, and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:21. CD138 peptides from Group B include peptides having an amino acid sequence from SEQ ID NO: 21 that comprises part or all of any one of SEQ ID NOS: 31 and 40-45, e.g., a sequence that comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids N-terminal and/or C-terminal to any one of SEQ ID NOS: 31 and 46-50. The amino acid positions referred to in Table 2 are based on SEQ ID NO: 21.

[0217] "CS-1" peptides from Group B include those peptides depicted in Table 2 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) consecutive amino acids from the human CS-1 protein having 335 amino acids and the amino acid sequence of SEQ ID NO:22, and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:22. CS1 peptides from Group B include peptides having an amino acid sequence from SEQ ID NO: 22 that comprises part or all of any one of SEQ ID NOS: 32 and 46-50, e.g., a sequence that comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids N-terminal and/or C-terminal to any one of SEQ ID NOS: 32 and 46-50. The amino acid positions referred to in Table 2 are based on SEQ ID NO: 22.

[0218] Group C Peptides.

[0219] The disclosure features isolated peptides ("Group C peptides") comprising an amino acid sequence that has sufficient identity with or is identical to any one of SEQ ID NOS: 51-536 as depicted in Table 3.

TABLE-US-00003 TABLE 3 Examples of Group C peptides Protein of Origin Amino Acid Position Amino Acid Sequence SEQ ID NO non-spliced XBP1 180-194 LSPLQNISPWILAVL 51 non-spliced XBP1 180-195 LSPLQNISPWILAVLT 52 non-spliced XBP1 180-196 LSPLQNISPWILAVLTL 53 non-spliced XBP1 180-197 LSPLQNISPWILAVLTLQ 54 non-spliced XBP1 180-198 LSPLQNISPWILAVLTLQI 55 non-spliced XBP1 180-199 LSPLQNISPWILAVLTLQIQ 56 non-spliced XBP1 180-200 LSPLQNISPWILAVLTLQIQS 57 non-spliced XBP1 180-201 LSPLQNISPWILAVLTLQIQSL 58 non-spliced XBP1 180-202 LSPLQNISPWILAVLTLQIQSLI 59 non-spliced XBP1 180-203 LSPLQNISPWILAVLTLQIQSLIS 60 non-spliced XBP1 180-204 LSPLQNISPWILAVLTLQIQSLISC 61 non-spliced XBP1 180-205 LSPLQNISPWILAVLTLQIQSLISCW 62 non-spliced XBP1 180-206 LSPLQNISPWILAVLTLQIQSLISCWA 63 non-spliced XBP1 181-194 SPLQNISPWILAVL 64 non-spliced XBP1 181-195 SPLQNISPWILAVLT 65 non-spliced XBP1 181-196 SPLQNISPWILAVLTL 66 non-spliced XBP1 181-197 SPLQNISPWILAVLTLQ 67 non-spliced XBP1 181-198 SPLQNISPWILAVLTLQI 68 non-spliced XBP1 181-199 SPLQNISPWILAVLTLQIQ 69 non-spliced XBP1 181-200 SPLQNISPWILAVLTLQIQS 70 non-spliced XBP1 181-201 SPLQNISPWILAVLTLQIQSL 71 non-spliced XBP1 181-202 SPLQNISPWILAVLTLQIQSLI 72 non-spliced XBP1 181-203 SPLQNISPWILAVLTLQIQSLIS 73 non-spliced XBP1 181-204 SPLQNISPWILAVLTLQIQSLISC 74 non-spliced XBP1 181-205 SPLQNISPWILAVLTLQIQSLISCW 75 non-spliced XBP1 181-206 SPLQNISPWILAVLTLQIQSLISCWA 76 non-spliced XBP1 182-194 PLQNISPWILAVL 77 non-spliced XBP1 182-195 PLQNISPWILAVLT 78 non-spliced XBP1 182-196 PLQNISPWILAVLTL 79 non-spliced XBP1 182-197 PLQNISPWILAVLTLQ 80 non-spliced XBP1 182-198 PLQNISPWILAVLTLQI 81 non-spliced XBP1 182-199 PLQNISPWILAVLTLQIQ 82 non-spliced XBP1 182-200 PLQNISPWILAVLTLQIQS 83 non-spliced XBP1 182-201 PLQNISPWILAVLTLQIQSL 84 non-spliced XBP1 182-202 PLQNISPWILAVLTLQIQSLI 85 non-spliced XBP1 182-203 PLQNISPWILAVLTLQIQSLIS 86 non-spliced XBP1 182-204 PLQNISPWILAVLTLQIQSLISC 87 non-spliced XBP1 182-205 PLQNISPWILAVLTLQIQSLISCW 88 non-spliced XBP1 182-206 PLQNISPWILAVLTLQIQSLISCWA 89 non-spliced XBP1 183-194 LQNISPWILAVL 90 non-spliced XBP1 183-195 LQNISPWILAVLT 91 non-spliced XBP1 183-196 LQNISPWILAVLTL 92 non-spliced XBP1 183-197 LQNISPWILAVLTLQ 93 non-spliced XBP1 183-198 LQNISPWILAVLTLQI 94 non-spliced XBP1 183-199 LQNISPWILAVLTLQIQ 95 non-spliced XBP1 183-200 LQNISPWILAVLTLQIQS 96 non-spliced XBP1 183-201 LQNISPWILAVLTLQIQSL 97 non-spliced XBP1 183-202 LQNISPWILAVLTLQIQSLI 98 non-spliced XBP1 183-203 LQNISPWILAVLTLQIQSLIS 99 non-spliced XBP1 183-204 LQNISPWILAVLTLQIQSLISC 100 non-spliced XBP1 183-205 LQNISPWILAVLTLQIQSLISCW 101 non-spliced XBP1 183-206 LQNISPWILAVLTLQIQSLISCWA 102 non-spliced XBP1 184-194 QNISPWILAVL 103 non-spliced XBP1 184-195 QNISPWILAVLT 104 non-spliced XBP1 184-196 QNISPWILAVLTL 105 non-spliced XBP1 184-197 QNISPWILAVLTLQ 106 non-spliced XBP1 184-198 QNISPWILAVLTLQI 107 non-spliced XBP1 184-199 QNISPWILAVLTLQIQ 108 non-spliced XBP1 184-200 QNISPWILAVLTLQIQS 109 non-spliced XBP1 184-201 QNISPWILAVLTLQIQSL 110 non-spliced XBP1 184-202 QNISPWILAVLTLQIQSLI 111 non-spliced XBP1 184-203 QNISPWILAVLTLQIQSLIS 112 non-spliced XBP1 184-204 QNISPWILAVLTLQIQSLISC 113 non-spliced XBP1 184-205 QNISPWILAVLTLQIQSLISCW 114 non-spliced XBP1 184-206 QNISPWILAVLTLQIQSLISCWA 115 non-spliced XBP1 185-194 NISPWILAVL 116 non-spliced XBP1 185-195 NISPWILAVLT 117 non-spliced XBP1 185-196 NISPWILAVLTL 118 non-spliced XBP1 185-197 NISPWILAVLTLQ 119 non-spliced XBP1 185-198 NISPWILAVLTLQI 120 non-spliced XBP1 185-199 NISPWILAVLTLQIQ 121 non-spliced XBP1 185-200 NISPWILAVLTLQIQS 122 non-spliced XBP1 185-201 NISPWILAVLTLQIQSL 123 non-spliced XBP1 185-202 NISPWILAVLTLQIQSLI 124 non-spliced XBP1 185-203 NISPWILAVLTLQIQSLIS 125 non-spliced XBP1 185-204 NISPWILAVLTLQIQSLISC 126 non-spliced XBP1 185-205 NISPWILAVLTLQIQSLISCW 127 non-spliced XBP1 185-206 NISPWILAVLTLQIQSLISCWA 128 non-spliced XBP1 185-194 YISPWILAVL 129 non-spliced XBP1 185-195 YISPWILAVLT 130 non-spliced XBP1 185-196 YISPWILAVLTL 131 non-spliced XBP1 185-197 YISPWILAVLTLQ 132 non-spliced XBP1 185-198 YISPWILAVLTLQI 133 non-spliced XBP1 185-199 YISPWILAVLTLQIQ 134 non-spliced XBP1 185-200 YISPWILAVLTLQIQS 135 non-spliced XBP1 185-201 YISPWILAVLTLQIQSL 136 non-spliced XBP1 185-202 YISPWILAVLTLQIQSLI 137 non-spliced XBP1 185-203 YISPWILAVLTLQIQSLIS 138 non-spliced XBP1 185-204 YISPWILAVLTLQIQSLISC 139 non-spliced XBP1 185-205 YISPWILAVLTLQIQSLISCW 140 non-spliced XBP1 185-206 YISPWILAVLTLQIQSLISCWA 141 non-spliced XBP1 186-194 ISPWILAVL 142 non-spliced XBP1 186-195 ISPWILAVLT 143 non-spliced XBP1 186-196 ISPWILAVLTL 144 non-spliced XBP1 186-197 ISPWILAVLTLQ 145 non-spliced XBP1 186-198 ISPWILAVLTLQI 146 non-spliced XBP1 186-199 ISPWILAVLTLQIQ 147 non-spliced XBP1 186-200 ISPWILAVLTLQIQS 148 non-spliced XBP1 186-201 ISPWILAVLTLQIQSL 149 non-spliced XBP1 186-202 ISPWILAVLTLQIQSLI 150 non-spliced XBP1 186-203 ISPWILAVLTLQIQSLIS 151 non-spliced XBP1 186-204 ISPWILAVLTLQIQSLISC 152 non-spliced XBP1 186-205 ISPWILAVLTLQIQSLISCW 153 non-spliced XBP1 186-206 ISPWILAVLTLQIQSLISCWA 154 non-spliced XBP1 187-194 SPWILAVL 155 non-spliced XBP1 187-195 SPWILAVLT 156 non-spliced XBP1 187-196 SPWILAVLTL 157 non-spliced XBP1 187-197 SPWILAVLTLQ 158 non-spliced XBP1 187-198 SPWILAVLTLQI 159 non-spliced XBP1 187-199 SPWILAVLTLQIQ 160 non-spliced XBP1 187-200 SPWILAVLTLQIQS 161 non-spliced XBP1 187-201 SPWILAVLTLQIQSL 162 non-spliced XBP1 187-202 SPWILAVLTLQIQSLI 163 non-spliced XBP1 187-203 SPWILAVLTLQISSLIS 164 non-spliced XBP1 187-204 SPWILAVLTLQISSLISC 165 non-spliced XBP1 187-205 SPWILAVLTLQISSLISCW 166 non-spliced XBP1 187-206 SPWILAVLTLQISSLISCWA 167 non-spliced XBP1 188-194 PWILAVL 168 non-spliced XBP1 188-195 PWILAVLT 169 non-spliced XBP1 188-196 PWILAVLTL 170 non-spliced XBP1 188-197 PWILAVLTLQ 171 non-spliced XBP1 188-198 PWILAVLTLQI 172 non-spliced XBP1 188-199 PWILAVLTLQIQ 173

non-spliced XBP1 188-200 PWILAVLTLQIQS 174 non-spliced XBP1 188-201 PWILAVLTLQIQSL 175 non-spliced XBP1 188-202 PWILAVLTLQIQSLI 176 non-spliced XBP1 188-203 PWILAVLTLQIQSLIS 177 non-spliced XBP1 188-204 PWILAVLTLQIQSLISC 178 non-spliced XBP1 188-205 PWILAVLTLQIQSLISCW 179 non-spliced XBP1 188-206 PWILAVLTLQIQSLISCWA 180 non-spliced XBP1 189-194 WILAVL 181 non-spliced XBP1 189-195 WILAVLT 182 non-spliced XBP1 189-196 WILAVLTL 183 non-spliced XBP1 189-197 WILAVLTLQ 184 non-spliced XBP1 189-198 WILAVLTLQI 185 non-spliced XBP1 189-199 WILAVLTLQIQ 186 non-spliced XBP1 189-200 WILAVLTLQIQS 187 non-spliced XBP1 189-201 WILAVLTLQIQSL 188 non-spliced XBP1 189-202 WILAVLTLQIQSLI 189 non-spliced XBP1 189-203 WILAVLTLQIQSLIS 190 non-spliced XBP1 189-204 WILAVLTLQIQSLISC 191 non-spliced XBP1 189-205 WILAVLTLQIQSLISCW 192 non-spliced XBP1 189-206 WILAVLTLQIQSLISCWA 193 non-spliced XBP1 190-194 ILAVL 194 non-spliced XBP1 190-195 ILAVLT 195 non-spliced XBP1 190-196 ILAVLTL 196 non-spliced XBP1 190-197 ILAVLTLQ 197 non-spliced XBP1 190-198 ILAVLTLQI 198 non-spliced XBP1 190-199 ILAVLTLQIQ 199 non-spliced XBP1 190-200 ILAVLTLQIQS 200 non-spliced XBP1 190-201 ILAVLTLQIQSL 201 non-spliced XBP1 190-202 ILAVLTLQIQSLI 202 non-spliced XBP1 190-203 ILAVLTLQIQSLIS 203 non-spliced XBP1 190-204 ILAVLTLQIQSLISC 204 non-spliced XBP1 190-205 ILAVLTLQIQSLISCW 205 non-spliced XBP1 190-206 ILAVLTLQIQSLISCWA 206 CD138 251-268 EVLGGVIAGGLVGLIFAV 207 CD138 251-269 EVLGGVIAGGLVGLIFAVC 208 CD138 251-270 EVLGGVIAGGLVGLIFAVCL 209 CD138 251-271 EVLGGVIAGGLVGLIFAVCLV 210 CD138 251-272 EVLGGVIAGGLVGLIFAVCLVG 211 CD138 251-273 EVLGGVIAGGLVGLIFAVCLVGF 212 CD138 251-274 EVLGGVIAGGLVGLIFAVCLVGFM 213 CD138 251-275 EVLGGVIAGGLVGLIFAVCLVGFML 214 CD138 251-276 EVLGGVIAGGLVGLIFAVCLVGFMLY 215 CD138 251-277 EVLGGVIAGGLVGLIFAVCLVGFMLYR 216 CD138 251-278 EVLGGVIAGGLVGLIFAVCLVGFMLYRM 217 CD138 252-268 VLGGVIAGGLVGLIFAV 218 CD138 252-269 VLGGVIAGGLVGLIFAVC 219 CD138 252-270 VLGGVIAGGLVGLIFAVCL 220 CD138 252-271 VLGGVIAGGLVGLIFAVCLV 221 CD138 252-272 VLGGVIAGGLVGLIFAVCLVG 222 CD138 252-273 VLGGVIAGGLVGLIFAVCLVGF 223 CD138 252-274 VLGGVIAGGLVGLIFAVCLVGFM 224 CD138 252-275 VLGGVIAGGLVGLIFAVCLVGFML 225 CD138 252-276 VLGGVIAGGLVGLIFAVCLVGFMLY 226 CD138 252-277 VLGGVIAGGLVGLIFAVCLVGFMLYR 227 CD138 252-278 VLGGVIAGGLVGLIFAVCLVGFMLYRM 228 CD138 253-268 LGGVIAGGLVGLIFAV 229 CD138 253-269 LGGVIAGGLVGLIFAVC 230 CD138 253-270 LGGVIAGGLVGLIFAVCL 231 CD138 253-271 LGGVIAGGLVGLIFAVCLV 232 CD138 253-272 LGGVIAGGLVGLIFAVCLVG 233 CD138 253-273 LGGVIAGGLVGLIFAVCLVGF 234 CD138 253-274 LGGVIAGGLVGLIFAVCLVGFM 235 CD138 253-275 LGGVIAGGLVGLIFAVCLVGFML 236 CD138 253-276 LGGVIAGGLVGLIFAVCLVGFMLY 237 CD138 253-277 LGGVIAGGLVGLIFAVCLVGFMLYR 238 CD138 253-278 LGGVIAGGLVGLIFAVCLVGFMLYRM 239 CD138 254-268 GGVIAGGLVGLIFAV 240 CD138 254-269 GGVIAGGLVGLIFAVC 241 CD138 254-270 GGVIAGGLVGLIFAVCL 242 CD138 254-271 GGVIAGGLVGLIFAVCLV 243 CD138 254-272 GGVIAGGLVGLIFAVCLVG 244 CD138 254-273 GGVIAGGLVGLIFAVCLVGF 245 CD138 254-274 GGVIAGGLVGLIFAVCLVGFM 246 CD138 254-275 GGVIAGGLVGLIFAVCLVGFML 247 CD138 254-276 GGVIAGGLVGLIFAVCLVGFMLY 248 CD138 254-277 GGVIAGGLVGLIFAVCLVGFMLYR 249 CD138 254-278 GGVIAGGLVGLIFAVCLVGFMLYRM 250 CD138 255-268 GVIAGGLVGLIFAV 251 CD138 255-269 GVIAGGLVGLIFAVC 252 CD138 255-270 GVIAGGLVGLIFAVCL 253 CD138 255-271 GVIAGGLVGLIFAVCLV 254 CD138 255-272 GVIAGGLVGLIFAVCLVG 255 CD138 255-273 GVIAGGLVGLIFAVCLVGF 256 CD138 255-274 GVIAGGLVGLIFAVCLVGFM 257 CD138 255-275 GVIAGGLVGLIFAVCLVGFML 258 CD138 255-276 GVIAGGLVGLIFAVCLVGFMLY 259 CD138 255-277 GVIAGGLVGLIFAVCLVGFMLYR 260 CD138 255-278 GVIAGGLVGLIFAVCLVGFMLYRM 261 CD138 256-268 VIAGGLVGLIFAV 262 CD138 256-269 VIAGGLVGLIFAVC 263 CD138 256-270 VIAGGLVGLIFAVCL 264 CD138 256-271 VIAGGLVGLIFAVCLV 265 CD138 256-272 VIAGGLVGLIFAVCLVG 266 CD138 256-273 VIAGGLVGLIFAVCLVGF 267 CD138 256-274 VIAGGLVGLIFAVCLVGFM 268 CD138 256-275 VIAGGLVGLIFAVCLVGFML 269 CD138 256-276 VIAGGLVGLIFAVCLVGFMLY 270 CD138 256-277 VIAGGLVGLIFAVCLVGFMLYR 271 CD138 256-278 VIAGGLVGLIFAVCLVGFMLYRM 272 CD138 257-268 IAGGLVGLIFAV 273 CD138 257-269 IAGGLVGLIFAVC 274 CD138 257-270 IAGGLVGLIFAVCL 275 CD138 257-271 IAGGLVGLIFAVCLV 276 CD138 257-272 IAGGLVGLIFAVCLVG 277 CD138 257-273 IAGGLVGLIFAVCLVGF 278 CD138 257-274 IAGGLVGLIFAVCLVGFM 279 CD138 257-275 IAGGLVGLIFAVCLVGFML 280 CD138 257-276 IAGGLVGLIFAVCLVGFMLY 281 CD138 257-277 IAGGLVGLIFAVCLVGFMLYR 282 CD138 257-278 IAGGLVGLIFAVCLVGFMLYRM 283 CD138 258-268 AGGLVGLIFAV 284 CD138 258-269 AGGLVGLIFAVC 285 CD138 258-270 AGGLVGLIFAVCL 286 CD138 258-271 AGGLVGLIFAVCLV 287 CD138 258-272 AGGLVGLIFAVCLVG 288 CD138 258-273 AGGLVGLIFAVCLVGF 289 CD138 258-274 AGGLVGLIFAVCLVGFM 290 CD138 258-275 AGGLVGLIFAVCLVGFML 291 CD138 258-276 AGGLVGLIFAVCLVGFMLY 292 CD138 258-277 AGGLVGLIFAVCLVGFMLYR 293 CD138 258-278 AGGLVGLIFAVCLVGFMLYRM 294 CD138 259-268 GGLVGLIFAV 295 CD138 259-269 GGLVGLIFAVC 296 CD138 259-270 GGLVGLIFAVCL 297 CD138 259-271 GGLVGLIFAVCLV 298

CD138 259-272 GGLVGLIFAVCLVG 299 CD138 259-273 GGLVGLIFAVCLVGF 300 CD138 259-274 GGLVGLIFAVCLVGFM 301 CD138 259-275 GGLVGLIFAVCLVGFML 302 CD138 259-276 GGLVGLIFAVCLVGFMLY 303 CD138 259-277 GGLVGLIFAVCLVGFMLYR 304 CD138 259-278 GGLVGLIFAVCLVGFMLYRM 305 CD138 260-268 GLVGLIFAV 306 CD138 260-269 GLVGLIFAVC 307 CD138 260-270 GLVGLIFAVCL 308 CD138 260-271 GLVGLIFAVCLV 309 CD138 260-272 GLVGLIFAVCLVG 310 CD138 260-273 GLVGLIFAVCLVGF 311 CD138 260-274 GLVGLIFAVCLVGFM 312 CD138 260-275 GLVGLIFAVCLVGFML 313 CD138 260-276 GLVGLIFAVCLVGFMLY 314 CD138 260-277 GLVGLIFAVCLVGFMLYR 315 CD138 260-278 GLVGLIFAVCLVGFMLYRM 316 CD138 261-268 LVGLIFAV 317 CD138 261-269 LVGLIFAVC 318 CD138 261-270 LVGLIFAVCL 319 CD138 261-271 LVGLIFAVCLV 320 CD138 260-272 LVGLIFAVCLVG 321 CD138 261-273 LVGLIFAVCLVGF 322 CD138 261-274 LVGLIFAVCLVGFM 323 CD138 261-275 LVGLIFAVCLVGFML 324 CD138 261-276 LVGLIFAVCLVGFMLY 325 CD138 261-277 LVGLIFAVCLVGFMLYR 326 CD138 261-278 LVGLIFAVCLVGFMLYRM 327 CD138 262-268 VGLIFAV 328 CD138 262-269 VGLIFAVC 329 CD138 262-270 VGLIFAVCL 330 CD138 262-271 VGLIFAVCLV 331 CD138 262-272 VGLIFAVCLVG 332 CD138 262-273 VGLIFAVCLVGF 333 CD138 262-274 VGLIFAVCLVGFM 334 CD138 262-275 VGLIFAVCLVGFML 335 CD138 262-276 VGLIFAVCLVGFMLY 336 CD138 262-277 VGLIFAVCLVGFMLYR 337 CD138 262-278 VGLIFAVCLVGFMLYRM 338 CD138 263-268 GLIFAV 339 CD138 263-269 GLIFAVC 340 CD138 263-270 GLIFAVCL 341 CD138 263-271 GLIFAVCLV 342 CD138 263-272 GLIFAVCLVG 343 CD138 263-273 GLIFAVCLVGF 344 CD138 263-274 GLIFAVCLVGFM 345 CD138 263-275 GLIFAVCLVGFML 346 CD138 263-276 GLIFAVCLVGFMLY 347 CD138 263-277 GLIFAVCLVGFMLYR 348 CD138 263-278 GLIFAVCLVGFMLYRM 349 CD138 264-268 LIFAV 350 CD138 264-269 LIFAVC 351 CD138 264-270 LIFAVCL 352 CD138 264-271 LIFAVCLV 353 CD138 264-272 LIFAVCLVG 354 CD138 264-273 LIFAVCLVGF 355 CD138 264-274 LIFAVCLVGFM 356 CD138 264-275 LIFAVCLVGFML 357 CD138 264-276 LIFAVCLVGFMLY 358 CD138 264-277 LIFAVCLVGFMLYR 359 CD138 264-278 LIFAVCLVGFMLYRM 360 CD138 265-268 IFAV 361 CD138 265-269 IFAVC 362 CD138 265-270 IFAVCL 363 CD138 265-271 IFAVCLV 364 CD138 265-272 IFAVCLVG 365 CD138 265-273 IFAVCLVGF 366 CD138 265-274 IFAVCLVGFM 367 CD138 265-275 IFAVCLVGFML 368 CD138 265-276 IFAVCLVGFMLY 369 CD138 265-277 IFAVCLVGFMLYR 370 CD138 265-278 IFAVCLVGFMLYRM 371 CS1 227-242 VLLCLLLVPLLLSLFV 372 CS1 227-243 VLLCLLLVPLLLSLFVL 373 CS1 227-244 VLLCLLLVPLLLSLFVLG 374 CS1 227-245 VLLCLLLVPLLLSLFVLGL 375 CS1 227-246 VLLCLLLVPLLLSLFVLGLF 376 CS1 227-247 VLLCLLLVPLLLSLFVLGLFL 377 CS1 227-248 VLLCLLLVPLLLSLFVLGLFLW 378 CS1 227-249 VLLCLLLVPLLLSLFVLGLFLWF 379 CS1 227-250 VLLCLLLVPLLLSLFVLGLFLWFL 380 CS1 227-251 VLLCLLLVPLLLSLFVLGLFLWFLK 381 CS1 227-252 VLLCLLLVPLLLSLFVLGLFLWFLKR 382 CS1 227-253 VLLCLLLVPLLLSLFVLGLFLWFLKRE 383 CS1 228-242 LLCLLLVPLLLSLFV 384 CS1 228-243 LLCLLLVPLLLSLFVL 385 CS1 228-244 LLCLLLVPLLLSLFVLG 386 CS1 228-245 LLCLLLVPLLLSLFVLGL 387 CS1 228-246 LLCLLLVPLLLSLFVLGLF 388 CS1 228-247 LLCLLLVPLLLSLFVLGLFL 389 CS1 228-248 LLCLLLVPLLLSLFVLGLFLW 390 CS1 228-249 LLCLLLVPLLLSLFVLGLFLWF 391 CS1 228-250 LLCLLLVPLLLSLFVLGLFLWFL 392 CS1 228-251 LLCLLLVPLLLSLFVLGLFLWFLK 393 CS1 228-252 LLCLLLVPLLLSLFVLGLFLWFLKR 394 CS1 228-253 LLCLLLVPLLLSLFVLGLFLWFLKRE 395 CS1 229-242 LCLLLVPLLLSLFV 396 CS1 229-243 LCLLLVPLLLSLFVL 397 CS1 229-244 LCLLLVPLLLSLFVLG 398 CS1 229-245 LCLLLVPLLLSLFVLGL 399 CS1 229-246 LCLLLVPLLLSLFVLGLF 400 CS1 229-247 LCLLLVPLLLSLFVLGLFL 401 CS1 229-248 LCLLLVPLLLSLFVLGLFLW 402 CS1 229-249 LCLLLVPLLLSLFVLGLFLWF 403 CS1 229-250 LCLLLVPLLLSLFVLGLFLWFL 404 CS1 229-251 LCLLLVPLLLSLFVLGLFLWFLK 405 CS1 229-252 LCLLLVPLLLSLFVLGLFLWFLKR 406 CS1 229-253 LCLLLVPLLLSLFVLGLFLWFLKRE 407 CS1 230-242 CLLLVPLLLSLFV 408 CS1 230-243 CLLLVPLLLSLFVL 409 CS1 230-244 CLLLVPLLLSLFVLG 410 CS1 230-245 CLLLVPLLLSLFVLGL 411 CS1 230-246 CLLLVPLLLSLFVLGLF 412 CS1 230-247 CLLLVPLLLSLFVLGLFL 413 CS1 230-248 CLLLVPLLLSLFVLGLFLW 414 CS1 230-249 CLLLVPLLLSLFVLGLFLWF 415 CS1 230-250 CLLLVPLLLSLFVLGLFLWFL 416 CS1 230-251 CLLLVPLLLSLFVLGLFLWFLK 417 CS1 230-252 CLLLVPLLLSLFVLGLFLWFLKR 418 CS1 230-253 CLLLVPLLLSLFVLGLFLWFLKRE 419 CS1 231-242 LLLVPLLLSLFV 420 CS1 231-243 LLLVPLLLSLFVL 421 CS1 231-244 LLLVPLLLSLFVLG 422 CS1 231-245 LLLVPLLLSLFVLGL 423 CS1 231-246 LLLVPLLLSLFVLGLF 424

CS1 231-247 LLLVPLLLSLFVLGLFL 425 CS1 231-248 LLLVPLLLSLFVLGLFLW 426 CS1 231-249 LLLVPLLLSLFVLGLFLWF 427 CS1 231-250 LLLVPLLLSLFVLGLFLWFL 428 CS1 231-251 LLLVPLLLSLFVLGLFLWFLK 429 CS1 231-252 LLLVPLLLSLFVLGLFLWFLKR 430 CS1 231-253 LLLVPLLLSLFVLGLFLWFLKRE 431 CS1 232-242 LLVPLLLSLFV 432 CS1 232-243 LLVPLLLSLFVL 433 CS1 232-244 LLVPLLLSLFVLG 434 CS1 232-245 LLVPLLLSLFVLGL 435 CS1 232-246 LLVPLLLSLFVLGLF 436 CS1 232-247 LLVPLLLSLFVLGLFL 437 CS1 232-248 LLVPLLLSLFVLGLFLW 438 CS1 232-249 LLVPLLLSLFVLGLFLWF 439 CS1 232-250 LLVPLLLSLFVLGLFLWFL 440 CS1 232-251 LLVPLLLSLFVLGLFLWFLK 441 CS1 232-252 LLVPLLLSLFVLGLFLWFLKR 442 CS1 232-253 LLVPLLLSLFVLGLFLWFLKRE 443 CS1 233-242 LVPLLLSLFV 444 CS1 233-243 LVPLLLSLFVL 445 CS1 233-244 LVPLLLSLFVLG 446 CS1 233-245 LVPLLLSLFVLGL 447 CS1 233-246 LVPLLLSLFVLGLF 448 CS1 233-247 LVPLLLSLFVLGLFL 449 CS1 233-248 LVPLLLSLFVLGLFLW 450 CS1 233-249 LVPLLLSLFVLGLFLWF 451 CS1 233-250 LVPLLLSLFVLGLFLWFL 452 CS1 233-251 LVPLLLSLFVLGLFLWFLK 453 CS1 233-252 LVPLLLSLFVLGLFLWFLKR 454 CS1 233-253 LVPLLLSLFVLGLFLWFLKRE 455 CS1 234-242 VPLLLSLFV 456 CS1 234-243 VPLLLSLFVL 457 CS1 234-244 VPLLLSLFVLG 458 CS1 234-245 VPLLLSLFVLGL 459 CS1 234-246 VPLLLSLFVLGLF 460 CS1 234-247 VPLLLSLFVLGLFL 461 CS1 234-248 VPLLLSLFVLGLFLW 462 CS1 234-249 VPLLLSLFVLGLFLWF 463 CS1 234-250 VPLLLSLFVLGLFLWFL 464 CS1 234-251 VPLLLSLFVLGLFLWFLK 465 CS1 234-252 VPLLLSLFVLGLFLWFLKR 466 CS1 234-253 VPLLLSLFVLGLFLWFLKRE 467 CS1 235-242 PLLLSLFV 468 CS1 235-243 PLLLSLFVL 469 CS1 235-244 PLLLSLFVLG 470 CS1 235-245 PLLLSLFVLGL 471 CS1 235-246 PLLLSLFVLGLF 472 CS1 235-247 PLLLSLFVLGLFL 473 CS1 235-248 PLLLSLFVLGLFLW 474 CS1 235-249 PLLLSLFVLGLFLWF 475 CS1 235-250 PLLLSLFVLGLFLWFL 476 CS1 235-251 PLLLSLFVLGLFLWFLK 477 CS1 235-252 PLLLSLFVLGLFLWFLKR 478 CS1 235-253 PLLLSLFVLGLFLWFLKRE 479 CS1 236-242 LLLSLFV 480 CS1 236-243 LLLSLFVL 481 CS1 236-244 LLLSLFVLG 482 CS1 236-245 LLLSLFVLGL 483 CS1 236-246 LLLSLFVLGLF 484 CS1 236-247 LLLSLFVLGLFL 485 CS1 236-248 LLLSLFVLGLFLW 486 CS1 236-249 LLLSLFVLGLFLWF 487 CS1 236-250 LLLSLFVLGLFLWFL 488 CS1 236-251 LLLSLFVLGLFLWFLK 489 CS1 236-252 LLLSLFVLGLFLWFLKR 490 CS1 236-253 LLLSLFVLGLFLWFLKRE 491 CS1 237-242 LLSLFV 492 CS1 237-243 LLSLFVL 493 CS1 237-244 LLSLFVLG 494 CS1 237-245 LLSLFVLGL 495 CS1 237-246 LLSLFVLGLF 496 CS1 237-247 LLSLFVLGLFL 497 CS1 237-248 LLSLFVLGLFLW 498 CS1 237-249 LLSLFVLGLFLWF 499 CS1 237-250 LLSLFVLGLFLWFL 500 CS1 237-251 LLSLFVLGLFLWFLK 501 CS1 237-252 LLSLFVLGLFLWFLKR 502 CS1 237-253 LLSLFVLGLFLWFLKRE 503 CS1 238-242 LSLFV 504 CS1 238-243 LSLFVL 505 CS1 238-244 LSLFVLG 506 CS1 238-245 LSLFVLGL 507 CS1 238-246 LSLFVLGLF 508 CS1 238-247 LSLFVLGLFL 509 CS1 238-248 LSLFVLGLFLW 510 CS1 238-249 LSLFVLGLFLWF 511 CS1 238-250 LSLFVLGLFLWFL 512 CS1 238-251 LSLFVLGLFLWFLK 513 CS1 238-252 LSLFVLGLFLWFLKR 514 CS1 238-253 LSLFVLGLFLWFLKRE 515 CS1 239-243 SLFVL 516 CS1 239-244 SLFVLG 517 CS1 239-245 SLFVLGL 518 CS1 239-246 SLFVLGLF 519 CS1 239-247 SLFVLGLFL 520 CS1 239-248 SLFVLGLFLW 521 CS1 239-249 SLFVLGLFLWF 522 CS1 239-250 SLFVLGLFLWFL 523 CS1 239-251 SLFVLGLFLWFLK 524 CS1 239-252 SLFVLGLFLWFLKR 525 CS1 239-253 SLFVLGLFLWFLKRE 526 CS1 240-244 LFVLG 527 CS1 240-245 LFVLGL 528 CS1 240-246 LFVLGLF 529 CS1 240-247 LFVLGLFL 530 CS1 240-248 LFVLGLFLW 531 CS1 240-249 LFVLGLFLWF 532 CS1 240-250 LFVLGLFLWFL 533 CS1 240-251 LFVLGLFLWFLK 534 CS1 240-252 LFVLGLFLWFLKR 535 CS1 240-253 LFVLGLFLWFLKRE 536

[0220] Preferably, the isolated peptide from Group C is at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or 35 amino acids in length (e.g., between 9 and 35 amino acids in length, e.g., 9-30, 9-25, 9-20, 9-15 amino acids in length) and comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or is identical to an amino acid sequence of SEQ ID NOS: 51-536. Other preferred peptides can be at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or 35 amino acids in length (e.g., between 9 and 35 amino acids in length, e.g., 9-30, 9-25, 9-20, 9-15 amino acids in length) and comprise an amino acid sequence of SEQ ID NOs: 51-536, or an amino acid sequence with one, two, three or four substitutions of the amino acid sequence of SEQ ID NOs: 51-536. The substitution can be a conservative or nonconservative substitution.

[0221] "Non-spliced XBP1" peptides from Group C include those peptides depicted in Table 3 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) consecutive amino acids from the non-spliced form of human XBP1 protein having 261 amino acids and the amino acid sequence of SEQ ID NO:19, and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:19. Non-spliced XBP1 peptides from Group C include peptides having an amino acid sequence from SEQ ID NO: 19 that comprises part or all of SEQ ID NOS: 51-206. The amino acid positions referred to in Table 3 are based on SEQ ID NO: 19.

[0222] "CD138" peptides from Group C include those peptides depicted in Table 3 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) consecutive amino acids from the human CD138 protein having 310 amino acids and the amino acid sequence of SEQ ID NO:21, and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:21. CD138 peptides from Group C include peptides having an amino acid sequence from SEQ ID NO: 21 that comprises part or all of SEQ ID NOS: 207-371.

[0223] The amino acid positions referred to in Table 3 are based on SEQ ID NO: 21.

[0224] "CS-1" peptides from Group C include those peptides depicted in Table 3 and refer to a peptide having an amino acid sequence of at least 5 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) consecutive amino acids from the human CS-1 protein having 335 amino acids and the amino acid sequence of SEQ ID NO:22, and peptides having no more than one, two, three, four, five substitutions (e.g., conservative substitutions) of the amino acids derived from the amino acid sequence of SEQ ID NO:22. CS-1 peptides from Group C include peptides having an amino acid sequence from SEQ ID NO: 22 that comprises part or all of SEQ ID NOS: 372-536. The amino acid positions referred to in Table 3 are based on SEQ ID NO: 22.

[0225] Peptides Generally.

[0226] The peptides described herein are often referred to using the residue number of the N and C terminal amino acids of the peptides (e.g., XBP1.sub.118-126) as the relevant sequences occur in the wild-type, full length, mature human proteins having SEQ ID NOS: 19-22. These peptides will frequently have identical sequences to the corresponding segments of the wild-type, full-length, mature proteins having SEQ ID NOS: 19-22. It is understood, however, that the terms "nonspliced XBP1 peptides" (e.g., nonspliced XBP1 peptides having amino acid positions: 118-136, 185-193, 186-194, 190-198, 193-200, or 111-119), "spliced XBP1 peptides" (e.g., spliced XBP1 peptides having amino acid positions: 197-205, 194-202, 224-232, 368-376), "CD138 peptides" (e.g., CD138 peptides having amino acid positions: 256-264, 265-273, 260-268, 5-13, or 7-15), and CS1 peptides (e.g., CS-1 peptides have amino acid positions 236-245, 240-248, 239-247, 232-240, or 9-17) can be peptide fragments of the XBP1 nonspliced peptide, the XBP1 spliced peptide, the CD138, or CS-1 polypeptide (respectively) of a species other than human. As will be appreciated by those skilled in the art, the numbers of the N and C terminal amino acids of peptide fragments of such non-human polypeptides are not necessarily the same as those in the corresponding peptide fragments of human polypeptides. Moreover, the lengths and/or amino acids of peptide fragments of non-human polypeptides will not necessarily be the same as those in the corresponding peptide fragments of human polypeptides. Those of skill in the art will know how to establish the N and C terminal amino acids, the lengths, and amino acid sequences of peptides derived from non-human nonspliced XBP1, spliced XBP1, CD138, and CS-1 polypeptides. One useful method for doing this is sequence alignment and, in particular, maximum homology sequence alignment.

[0227] Percent identity between two peptide sequences (e.g., a peptide of SEQ ID NOS: 1-18 and 29-536) and another amino acid sequence that may be at least 66% identical to the peptide) can be determined using a variety of algorithms and computer programs including, but not limited to, Clustal W (The European Bioinformatics Institute (EMBL-EBI), BLAST-Protein (National Center for Biotechnology Information (NCBI), United States National Institutes of Health), and PSAlign (University of Texas A&M; Sze et al. (2006) Journal of Computational Biology 13:309-319).

[0228] Also disclosed herein are variants of the human and non-human peptides described above. Variants of the human and non-human peptides described herein can include forms of the peptides having: (i) not more than 4 (e.g., 3, 2, or 1) amino acid substitutions (e.g., conservative or non-conservative substitutions); (ii) terminal or internal deletions; or (iii) terminal or internal additions, all of which are elaborated on below.

[0229] The disclosure also features peptides comprising, consisting of, or consisting essentially of, an amino acid sequence of any of SEQ ID NOs: 1-18 and 29-536 (as depicted in Tables 1-3), but with not more than four (e.g., not more than three, not more than two, or not more than 1) substitutions. The substitutions can be, e.g., conservative or non-conservative (as described above).

[0230] Conservative substitutions include substitutions within the following groups: valine, alanine and glycine; leucine, valine, and isoleucine; aspartic acid and glutamic acid; asparagine and glutamine; serine, cysteine, and threonine; lysine and arginine; and phenylalanine and tyrosine. The non-polar hydrophobic amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and methionine. The polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine and glutamine. The positively charged (basic) amino acids include arginine, lysine, and histidine. The negatively charged (acidic) amino acids include aspartic acid and glutamic acid. Any substitution of one member of the above-mentioned polar, basic or acidic groups by another member of the same group can be deemed a conservative substitution. By contrast, a non-conservative substitution is a substitution of one amino acid for another with dissimilar characteristics.

[0231] In some embodiments, one or more (e.g., one, two, three, four, or all five) of positions three, four, five, six, seven, and eight of any of the peptides are not substituted. In some embodiments, one or more of positions three, four, five, six, seven, and eight of any of the peptides are identical to the amino acids of the peptides in Tables 1-3.

[0232] Also featured are fusion proteins comprising: a first amino acid sequence of a peptide described herein (e.g., a nonspliced XBP1 peptide described herein, a spliced XBP1 peptide described herein, a CD138 peptide described herein and/or a CS-1 peptide described herein); and a second amino acid sequence that is heterologous to the first amino acid sequence.

[0233] The second, heterologous amino acid sequence(s) of the peptide generally do not (and are selected such that do not) adversely affect the generation in the cell of an immunogenic peptide of any of SEQ ID NOs: 1-18 and 29-536. The cellular machinery is expected to remove any additional sequences in the peptide to yield an immunogenic peptide of any of SEQ ID NOs: 1-18 and 29-536, which peptide is presented by a class I or class II MHC molecule to stimulate an immune response against XBP1-, CD138-, or CS1-expressing cancer cells.

[0234] An amino acid sequence that is "heterologous" to a first amino acid sequence, or the term "heterologous amino acid sequence," is any amino acid sequence other than the amino acid sequence(s) flanking the first amino acid sequence as it occurs in nature. For example, two or more (e.g., two, three, four, five, six, seven, eight, nine, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more) and/or less than 20 (e.g., 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, nine, eight, seven, six, five, four, three, two, or one) carboxy- and/or amino-terminal amino acid(s) immediately flanking LLREKTHGL (SEQ ID NO:1) in a human XBP1 are not considered to be heterologous to SEQ ID NO:1. It is understood that a fusion protein containing a first amino acid sequence that is less than 100% identical to, or contains from one to four conservative substitutions in, an amino acid sequence of any of SEQ ID NOs: 1-18 and 29-536, may not occur in nature at all.

[0235] In some embodiments, the second amino acid sequence can be a single amino acid. It is understood that an amino acid that is "heterologous" to a first amino acid sequence, or the term "heterologous amino acid," is any amino acid other than the amino acid(s) flanking the first amino acid sequence as it occurs in nature. For example, the two amino acid(s) immediately flanking LLREKTHGL (SEQ ID NO:1) in a human XBP1 are not considered to be heterologous to SEQ ID NO:1.

[0236] A heterologous sequence can be, for example, a sequence used for purification of the recombinant protein (e.g., FLAG, polyhistidine (e.g., hexahistidine) (SEQ ID NO: 544), hemagluttanin (HA), glutathione-S-transferase (GST), or maltose-binding protein (MBP)). Heterologous sequences can also be proteins useful as diagnostic or detectable markers, for example, luciferase, green fluorescent protein (GFP), or chloramphenicol acetyl transferase (CAT). In some embodiments, the fusion protein can contain a signal sequence from another protein such as a KDEL (SEQ ID NO:23) sequence or any other described herein. In some embodiments, the fusion protein can contain all or part of an immunoglobulin molecule (e.g., all or part of an immunoglobulin heavy chain constant region; see below). In some embodiments, the fusion protein can contain a therapeutic or immune-stimulating polypeptide (e.g., a T helper epitope (e.g., a PADRE epitope or a Tetanus Toxoid universal T helper cell epitope) or all or part of a cytokine or chemokine) and/or a carrier (e.g., KLH) useful, e.g., in eliciting an immune response (e.g., for antibody generation). In some embodiments, the fusion protein can contain one or more linkers, e.g., a linker comprising a peptide sequence (see below). The fusion protein can also contain a targeting polypeptide. Heterologous sequences can be of varying length and in some cases can be longer sequences than the first amino acid sequences to which the heterologous amino acid sequences are attached. It is understood that a fusion protein containing a first amino acid sequence and a second amino acid sequence that is heterologous to the first does not correspond in sequence to a naturally occurring protein.

[0237] Targeting polypeptides, as used herein, are polypeptides that target the moiety (or moieties) they are attached to (e.g., the first amino acid sequence) to specific tissues (e.g., to a lymph node) or cells (e.g., to an antigen presenting cell or other immune cell), or where in vitro, specific isolated molecules or molecular complexes. Targeting polypeptides can be, e.g., an antibody (immunoglobulin) or antigen binding fragment thereof or a ligand for a cell surface receptor. An antibody (or antigen-binding fragment thereof) can be, e.g., a monoclonal antibody, a polyclonal antibody, a humanized antibody, a fully human antibody, a single chain antibody, a chimeric antibody, or an Fab fragment, an F(ab').sub.2 fragment, an Fab' fragment, an Fv fragment, or an scFv fragment of an antibody. Antibody fragments that include, or are, Fc regions (with or without antigen-binding regions) can also be used to target the reagents to Fc receptor-expressing cells (e.g., antigen presenting cells such as interdigitating dendritic cells, macrophages, monocytes, or B cells). A ligand for a cell surface receptor can be, e.g., a chemokine, a cytokine (e.g., interleukins 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16), or a death receptor ligand (e.g., FasL or TNF.alpha.).

[0238] In some embodiments, the heterologous sequence can be, e.g., a "transportation sequence" that aids in the delivery of the peptide to the cell or to a specific compartment of a cell (e.g., the endoplasmic reticulum or Golgi apparatus). Transportation sequences can include, e.g., membrane translocating sequence, a transportan sequence, an antennapedia sequence, a cyclic integrin-binding peptide, and a Tat-mediated peptide, or modified versions thereof.

[0239] A linker, e.g., a linker peptide, can, directly or indirectly, connect the first amino acid sequence to one or more heterologous amino acid sequences. For example, a linker can connect the first amino acid sequence to a second amino acid sequence. A linker peptide can be, or contain, e.g., stretches of amino acids where at least four to six amino acids are glycine. (See, e.g., Mancebo et al. (1990) Mol. Cell. Biol. 10:2492-2502). A linker peptide can also be, or contain, six or more (e.g., seven, eight, nine, 10, 11, or 12 or more) histidine residues. The linker peptide can be, or contain, at least one (e.g., one, two, three, four, five, six, seven, or eight or more) protease cleavage site(s). The protease sites can be, e.g., a trypsin, a chymotrypin, or a factor Xa cleavage site. Such protease sites can be useful, e.g., to separate a first amino acid sequence from a heterologous sequence. For example, after expression and purification of a fusion protein containing a first amino acid sequence joined to a polyhistidine sequence (in this case used for purification) by a trypsin protease cleavage site, the polyhistidine sequence can be removed from first amino acid sequence by contacting the fusion protein with trypsin.

[0240] The first amino acid sequence and the second amino acid sequence can be associated with each other in a variety of ways. As used herein, "associated with" in the context of an interaction between two or more atoms or molecular units, includes any covalent or non-covalent bonding, or physical admixture, of two or more atoms or molecular units (e.g., a first amino acid sequence and a second amino acid sequence). The chemical nature of covalent bonds (two atoms sharing one or more pairs of valence electrons) are known in the art and include, e.g., disulfide bonds or peptide bonds. A non-covalent bond is a chemical bond between atoms or molecules that does not involve the sharing of pairs of valence electrons. For example, non-covalent interactions include, e.g., hydrophobic interactions, hydrogen-bonding interactions, ionic bonding, Van der Waals bonding, or dipole-dipole interactions. Examples of such non-covalent interactions include antibody-antigen complexing or binding pair interactions (interactions of a first and second member of a binding pair such as the interaction between streptavidin and biotin). It is understood that the term "associated with" (e.g., in the context of a first amino acid sequence and a second amino acid sequence) is thus coextensive with the term "comprising."

[0241] In some embodiments, the first amino acid sequence and second amino acid sequence can be encoded by (and expressed as fusion protein from) a single nucleic acid sequence. In some instances, the first amino acid sequence and second amino acid sequence can be encoded by two or more (e.g., three, four, five, or six or more) different nucleic acid sequences. For example, the first amino acid sequence can be encoded by a first nucleic acid sequence and the second amino acid sequence can be encoded by a second nucleic acid sequence (see below under "Nucleic Acids and Methods for Producing the Peptides").

[0242] When expressed or produced separately, a first amino acid sequence and a second amino acid sequence can be cross-linked together using any of a number of known chemical cross linkers. Examples of such chemical cross-linkers are those which link two amino acid residues via a linkage that includes a "hindered" disulfide bond. In these linkages, a disulfide bond within the cross-linking unit is protected (by hindering groups on either side of the disulfide bond) from reduction by the action, for example, of reduced glutathione or the enzyme disulfide reductase. One suitable chemical cross-linker, 4-succinimidyloxycarbonyl-.alpha.-methyl-.alpha.(2-pyridyldithio)toluene (SMPT), forms such a linkage between the two amino acid sequences utilizing a terminal lysine on one of the amino acid sequences and a terminal cysteine on the other. Heterobifunctional reagents which cross-link by a different coupling moiety on each amino acid sequence. In this way, the resulting "dimers" will be heterodimers (peptides containing the first and second amino acid sequences) rather than either homodimers (e.g., two first amino acid sequences or two second amino acid sequences) or a mixture of homodimers and heterodimers. Thus, the coupling moiety on a first amino acid sequence could be a cysteine residue and on the other a lysine residue. Other useful cross-linkers include, without limitation, chemicals that link two amino groups (e.g., N-5-Azido-2-nitrobenzoyloxysuccinimide), two sulfhydryl groups (e.g., 1,4-Bis-maleimidobutane) an amino group and a sulfhydryl group (e.g., m-Maleimidobenzoyl-N-hydroxysuccinimide ester), an amino group and a carboxyl group (e.g., 4-[p-Azidosalicylamido]butylamine), and an amino group and a guanadium group that is present in the side chain of arginine (e.g., p-Azidophenyl glyoxal monohydrate).

[0243] The coupling moieties will preferably be at the termini (C or N) of each amino acid sequence. They can be, as indicated above, a cysteine residue on each amino acid sequence, or a cysteine on one and a lysine on the other. Where they are two cysteine residues, cross-linking can be effected by, for example, exposing amino acid sequences to oxidizing conditions.

[0244] A fusion protein can contain a first amino acid sequence and a second amino acid sequence or the fusion protein can contain more than one (e.g., two, three, four, five, six, seven, or eight or more) additional heterologous amino acid sequences. The additional heterologous amino acid sequences can flank, or be joined to, the amino terminus and/or the carboxy-terminus of the first amino acid sequence.

[0245] Where more than two amino acid sequences are to be joined, at least one of the amino acid sequences can have more than one cross-linking moiety. For example, a first amino acid sequence can have a cross-linking moiety at the amino-terminus and carboxy-terminus. Such multimers can be constructed "sequentially." Thus, each amino acid sequence is joined to the next such that the terminal amino acid sequences in the chain only have one residue involved in an inter-domain (or inter-agent) bond while the "internal" amino acid sequence(s) each have two moieties involved in inter-domain bonds. Alternatively, one amino acid sequence (such as the first amino acid sequence) could be linked to multiple (e.g., 2, 3, 4, or 5) other amino acid sequences.

[0246] Also featured are peptide compositions comprising: a first component and a second component, wherein the first component is a peptide described herein. The second component can be, e.g., a heterologous amino acid sequence (as described above), any other antigenic peptide (e.g., a peptide other than those described herein, a detectable label (see below), a therapeutic agent, a diagnostic agent, or a prophylactic agent (see below). For example, a peptide composition can contain an amino acid sequence consisting of, or consisting essentially of, any of SEQ ID NOs: 1-18 and 29-536 and a detectable label such as a radionuclide.

[0247] It is understood that in some embodiments, a peptide described herein can have at the amino-terminal end and/or carboxy-terminal end up to 200 (e.g., one, two, three, four, five, six, seven, eight, nine, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200) amino acids that are heterologous.

[0248] The peptides described herein can bind to a major histocompatibility complex (MHC) molecule (e.g., an MHC class I molecule or an MHC class II molecule). The "Major Histocompatibility Complex" or "MHC" is a cluster of genes that plays a role in control of the cellular interactions responsible for physiologic immune responses. In humans, the MHC is known as the HLA complex (see, e.g., Paul et al., FUNDAMENTAL IMMUNOLOGY, 3.sup.rd Edition, Raven Press, New York, (1993) and Stites, et al., IMMUNOLOGY, 8.sup.th Edition, Lange Publishing, Los Altos, Calif. (1994)).

[0249] An "HLA supertype or family," as used herein, refers to sets of HLA molecules grouped on the basis of shared peptide-binding specificities. HLA class I molecules that share somewhat similar binding affinity for peptides bearing certain amino acid motifs are grouped into HLA supertypes. The terms HLA superfamily, HLA supertype family, HLA family, and HLA xx-like molecules (where xx denotes a particular HLA type), are synonyms. Types of HLA class I molecules include, e.g., HLA-A1, HLA-A2, HLA-A3, HLA-A24, HLA-B7, HLA-B27, HLA-B44, HLA-B58, or HLA-B62. Such HLA molecules are described in detail in U.S. Pat. No. 7,026,443, the entire disclosure of which is incorporated by reference in its entirety.

[0250] A peptide can bind to an MHC molecule with high affinity or intermediate affinity. As used herein, "high affinity" binding of a peptide to an HLA class I molecule is defined as a binding with a dissociation constant (K.sub.D) of less than 50 (e.g., 45, 40, 35, 30, 25, 20, 15, 10, 5, 1, 0.5, 0.1, or less than 0.05) nM. "Intermediate affinity" is a binding of a peptide to an HLA class I molecule with a K.sub.D of between about 50 nM and about 500 nM (e.g., 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 115, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, or 500 nM). "High affinity" binding of a peptide to HLA class II molecules is defined as binding with a K.sub.D of less than 100 (e.g., 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, 5, 1, 0.5, 0.1, or less than 0.05) nM. "Intermediate affinity" of a peptide for an HLA class II molecule is binding with a K.sub.D of between about 100 and about 1000 nM (e.g., 100, 110, 115, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, or 1000 nM). Methods for determining the binding affinity of a peptide and an MHC molecule are known in the art and set forth in the accompanying Examples. Suitable methods are also described in, e.g., U.S. Pat. No. 7,026,443.

[0251] The peptides described herein can also be, in association with an MHC molecule, recognized by an antigen specific T cell receptor on a T cell. A variety of suitable methods can be used to determine whether a peptide, in association with an MHC molecule, is recognized by a T cell receptor on a T cell. For example, peripheral blood lymphocytes (PBL) from normal subjects can be cultured with a test peptide in the presence of antigen presenting cells in vitro over a period of several weeks. T cells specific for the peptide become activated during this time and can be detected using, e.g., proliferation assays (carboxyfluoroscein succinimidyl ester (CFSE) assays or .sup.3H-thymidine assays), limiting dilution assays, cytotoxicity assays (e.g., calcein-release assays), or cytokine- (e.g., IFN.gamma.), lymphokine-, or .sup.51Cr-release assays (see, e.g., Wentworth, P. A. et al., Mol. Immunol. 32:603, 1995; Celis, E. et al., Proc. Natl. Acad. Sci. USA 91:2105, 1994; Tsai, V. et al., J. Immunol. 158:1796, 1997; Kawashima, I. et al., Human Immunol. 59:1, 1998, the disclosures of each of which are incorporated by reference in their entirety). A suitable in vivo method involves immunizing HLA transgenic mice, wherein peptides in adjuvant are administered subcutaneously to HLA transgenic mice and several weeks following immunization, splenocytes are removed and cultured in vitro in the presence of test peptide for approximately one week and peptide-specific T cells are detected using, e.g., a .sup.51Cr-release assay (see, e.g., Wentworth, P. A. et al., J. Immunol. 26:97, 1996; Wentworth, P. A. et al., Int. Immunol. 8:651, 1996; Alexander, J. et al., J. Immunol. 159:4753, 1997, the disclosures of each of which are incorporated by reference in their entirety). Suitable methods are also set forth in the accompanying Examples. For example, the activation of a T cell by a peptide (in the context of an MHC molecule) can be determined by IFN-.gamma. cytokine production, CD107.alpha. degranulation or calcein release cytotoxicity assays (see, e.g., Examples 13 and 14).

[0252] Additionally, direct quantification of antigen-specific T cells can be performed by staining T cells with detectably-labeled MHC complexes such as any of the MHC molecule multimer compositions described herein (see below) or HLA-I tetramers (e.g., as described in Altman, J. D. et al., Proc. Natl. Acad. Sci. USA 90:10330, 1993 and Altman, J. D. et al., Science 274:94, 1996, the disclosures of each of which are incorporated by reference in their entirety).

[0253] In some embodiments, the peptides can be modified (e.g., amino acids of the peptides can be substituted) in order to modulate (e.g., increase or decrease) one of more properties of the peptides. For example, one or more (e.g., two, three, or four) amino acids of one of the peptides depicted in Table 1 can be substituted in order to increase the affinity of the peptide for an MHC molecule. In some embodiments, an amino acid of one of the peptides described herein (e.g., a T cell Receptor contacting amino acid residue of the peptide) can be modified in order to enhance a binding interaction between a T cell receptor and the peptide (in the context of an MHC molecule). Such modified peptides are often referred to as "altered peptide ligands." (See, e.g., Kalergis et al. (2000) J Immunol. 165(1):280; Conlon et al. (2002) Science 1801; and International Publication No. WO02070003, the disclosure of each of which is incorporated by reference in their entirety).

[0254] Suitable methods for modifying the peptides as well as determining the effect of the modification are set forth in the accompanying Examples and are described in, e.g., Collins et al. (Immunological Reviews (1998) 163:151-160, the disclosure of which is incorporated by reference in its entirety).

[0255] Nucleic Acids and Methods for Producing the Peptides

[0256] The disclosure also features nucleic acid sequences (as well as nucleic acid vectors containing nucleic acid sequences) encoding, and methods for producing, one or more (e.g., one, two, three, four, five, six, seven, eight, nine, 10, 11, 12, 13, or 14) of any of the peptides described herein (or a fusion protein described herein). Such methods can include the steps of: optionally, providing a cell (or group of cells) comprising a nucleic acid vector containing a nucleic acid sequence encoding one of more of any of the peptides described herein (or a fusion protein described herein), the nucleic acid sequence being operably linked to an expression control sequence, and culturing the cell under conditions that permit the expression of the peptides (or fusion protein). The methods can also include the step of isolating the one or more peptides (or protein) from the cell, or from the medium in which the cell was cultured.

[0257] Suitable methods for constructing nucleic acid sequences and vectors (e.g., expression vectors) for recombinant expression of one or more of the peptides (or fusion proteins) described herein are well known to those skilled in the art and described in, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual Second Edition vol. 1, 2 and 3. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, N.Y., USA, November 1989, the disclosure of which is incorporated by reference in its entirety. The nucleic acids and vectors can be used, e.g., to express the peptides (or fusion proteins) in a wide variety of host cells including, e.g., a bacterial, a yeast, or a mammalian cell. The nucleic acids and vectors can also be used in, e.g., in vivo and ex vivo methods as described below.

[0258] The peptide-coding sequences (or fusion protein-coding sequences) can be operably-linked to promoter and/or enhancer elements that direct the expression of the peptides (or fusion proteins) encoded by the nucleic acids. Enhancers provide expression specificity in terms of time, location, and level. Unlike a promoter, an enhancer can function when located at variable distances from the transcription initiation site, provided a promoter is present. An enhancer can also be located downstream of the transcription initiation site or in an exon of the relevant gene. To bring a coding sequence under the control of a promoter, it is necessary to position the translation initiation site of the translational reading frame of the peptide between one and about fifty nucleotides downstream (3') of the promoter. Promoters of interest include, but are not limited to, the cytomegalovirus hCMV immediate early gene, the early or late promoters of SV40 adenovirus, the lac system, the trp system, the TAC system, the TRC system, the major operator and promoter regions of phage A, the control regions of fd coat protein, the promoter for 3 phosphoglycerate kinase, the promoters of acid phosphatase, and the promoters of the yeast .alpha. mating factors, the adenoviral E1b minimal promoter, or the thymidine kinase minimal promoter.

[0259] The peptide-coding sequences, fusion protein-coding sequences, or vectors containing the such sequences, can contain a leader sequence that encodes a signal peptide. The leader sequence can be at the 5' end of the sequence encoding one or more of the peptides or fusion proteins described herein. The signal peptide can be immediately N-terminal of a given peptide (or fusion protein) or can be separated from it by one or more (e.g., 2, 3, 4, 6, 8, 10, 15 or 20) amino acids, provided that the leader sequence is in frame with the nucleic acid sequence encoding the peptide or fusion protein. The signal peptide, which is generally cleaved from the peptide (or fusion protein) prior to secretion (unless of course the signal peptide directs the insertion of a trasmembrane protein), directs the peptide (or fusion protein) to which it is attached into the lumen of the host cell endoplasmic reticulum (ER) during translation and the peptide (or fusion protein) is then secreted, via secretory vesicles, into the environment of the host cell. Useful signal peptides include, e.g., native leader sequences of cytokines or growth factors, KDEL (SEQ ID NO:23), or any signal sequences described in, e.g., U.S. Pat. No. 5,827,516, the disclosure of which is incorporated herein by reference in its entirety.

[0260] In some embodiments, the 5' end of a peptide-coding sequence (or fusion protein-coding sequence) can include a non-native ATG "start sequence." That is, e.g., an ATG sequence can be added to a nucleic acid encoding a peptide (or fusion protein) to ensure that the peptide (or fusion protein) is properly transcribed and translated. Although a leader sequence generally includes an ATG start sequence, in embodiments where it does not, the ATG sequence can be added at the 5' end of a nucleic acid encoding the leader sequence.

[0261] Suitable methods for constructing peptide-coding sequences and expression vectors are well known to those skilled in the art and described in, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual Second Edition vol. 1, 2 and 3. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, N.Y., USA, November 1989; the disclosure of which is incorporated herein by reference in its entirety.

[0262] A recombinant vector can be introduced into a cell using a variety of methods, which methods can depend, at least in part, on the type of cell into which the nucleic acid is introduced. For example, bacterial cells can be transformed using methods such as electroporation or heat shock. Methods for transfecting yeast cells include, e.g., the spheroplast technique or the whole-cell lithium chloride yeast transformation method (see, e.g., U.S. Pat. No. 4,929,555; Hinnen et al. (1978) Proc. Nat. Acad. Sci. USA 75:1929; Ito et al. (1983) J. Bacteriol. 153:163; U.S. Pat. No. 4,879,231; and Sreekrishna et al. (1987) Gene 59:115, the disclosures of each of which are incorporated herein by reference in their entirety). Transfection of animal cells can feature, for example, the introduction of a vector to the cells using calcium phosphate, electroporation, heat shock, liposomes, or transfection reagents such as FUGENE.RTM. or LIPOFECTAMINE.RTM., or by contacting naked nucleic acid vectors with the cells in solution (see, e.g., Sambrook et al., supra).

[0263] Expression systems that can be used for small or large scale production of the peptides (or fusion proteins) described herein include, but are not limited to, microorganisms such as bacteria (for example, E. coli and B. subtilis) transformed with recombinant bacteriophage DNA, plasmid DNA, or cosmid DNA expression vectors; fungus (e.g., yeast (for example, Saccharomyces and Pichia)) transformed with recombinant yeast expression vectors; insect cell systems infected with recombinant virus expression vectors (for example, baculovirus); plant cell systems infected with recombinant virus expression vectors (for example, cauliflower mosaic virus (CaMV) and tobacco mosaic virus (TMV)) or transformed with recombinant plasmid expression vectors (for example, Ti plasmid); or mammalian cell systems (for example, COS, CHO, BHK, 293, VERO, HeLa, MDCK, WI38, and NIH 3T3 cells) harboring recombinant expression constructs containing promoters derived from the genome of mammalian cells (for example, the metallothionein promoter) or from mammalian viruses (for example, the adenovirus late promoter, a CMV promoter, an SV40 promoter, or the vaccinia virus 7.5K promoter). Also useful as host cells are primary or secondary cells obtained directly from a mammal, transfected with a plasmid vector or infected with a viral vector (e.g., viral vectors such as herpes viruses, retroviruses, vaccinia viruses, attenuated vaccinia viruses, canary pox viruses, adenoviruses and adeno-associated viruses, among others).

[0264] As described above, following the expression of any of the peptides (or fusion proteins) described herein, the peptides (or fusion proteins) can be isolated from the cultured cells, or from the media in which the cells were cultured, using standard techniques (see Sambrook et al., supra). Methods of isolating proteins are known in the art and include, e.g., liquid chromatography (e.g., HPLC), affinity chromatography (e.g., metal chelation or immunoaffinity chromatography), ion-exchange chromatography, hydrophobic-interaction chromatography, precipitation, or differential solubilization.

[0265] Smaller peptides (e.g., peptides having less than 200 (e.g., less than 175, less than 150, less than 125, less than 100, less than 90, less than 80, less than 70, or less than 60) amino acids) can be chemically synthesized by standard chemical means such as FMOC solid-phase synthesis (see Example 1).

[0266] The peptides (and fusion proteins) described herein can, but need not, be isolated. The term "isolated," as applied to any of the peptides (or fusion proteins) described herein, refers to a peptide, a fragment thereof, (or for compositions, a macromolecular complex), that has been separated or purified from components (e.g., proteins or other naturally-occurring biological or organic molecules) which naturally accompany it. It is understood that recombinant molecules (e.g., recombinant peptides) will always be "isolated." Typically, a peptide (or fragment or macromolecular complex) is isolated when it constitutes at least 60%, by weight, of the total molecules of the same type in a preparation, e.g., 60% of the total molecules of the same type in a sample. For example, a peptide described herein is considered isolated when it constitutes at least 60%, by weight, of the total protein in a preparation or sample. In some embodiments, a molecule in the preparation consists of at least 75%, at least 90%, or at least 99%, by weight, of the total molecules of the same type in a preparation.

[0267] Similarly, the peptide-coding sequences, fusion protein-coding sequence or vectors containing such sequences described herein can also be isolated. The term "isolated," as applied to any of the peptide-coding sequences, fusion protein coding sequences or vectors described herein, refers to a peptide-coding sequence, a fusion protein coding sequence or vector, a fragment thereof that has been separated or purified from components (e.g., nucleic acids, proteins, or other naturally-occurring biological or organic molecules) which naturally accompany it. It is understood that recombinant molecules (e.g., recombinant vectors or peptide-coding or fusion protein-coding sequences) will always be "isolated." Typically, a peptide-coding sequence, fusion protein-coding sequence or vector (or fragment thereof) is isolated when it constitutes at least 60%, by weight, of the total molecules of the same type in a preparation, e.g., 60% of the total molecules of the same type in a sample. For example, a peptide-coding sequence or vector described herein is considered isolated when it constitutes at least 60%, by weight, of the total nucleic acid in a preparation or sample. In some embodiments, a molecule in the preparation consists of at least 75%, at least 90%, or at least 99%, by weight, of the total molecules of the same type in a preparation.

[0268] In some embodiments, the isolated peptides, fusion proteins, peptide-coding sequences, fusion protein-coding sequences or vectors can be frozen, lyophilized, or immobilized and stored under appropriate conditions, which allow the molecules to retain activity (e.g., the ability of a peptide to bind to an MHC molecule such as an MHC class I molecule or the ability of a vector to support expression of a peptide in a cell).

[0269] Additional Processing of the Peptides

[0270] Following the expression or synthesis of any of the peptides (or fusion proteins) described herein, the peptides (or fusion proteins) can be further processed. The further processing can include chemical or enzymatic modifications to peptides (or fusion protein) or, in cases where the peptides (or fusion proteins) are modified, the processing can include enzymatic or chemical alterations of existing modifications, or both. The additional processing of the peptides can include the addition (covalent or non-covalent joining) of a heterologous amino acid sequence such as, but not limited to, any of the heterologous amino acid sequences described above. Enzymatic treatment can involve contacting a peptide with, e.g., one or more proteases, phosphatases, or kinases under conditions that allow the peptide to be modified. Enzymatic treatment can involve contacting a peptide with one or more enzymes (e.g., an oligosaccharyltransferase or a mannosidase) capable of glycosylating, or modifying the glycosylation of, the peptide.

[0271] The processing can include the addition of, e.g., a detectable label to a peptide. For example, a peptide can be detectably labeled with an enzyme (e.g., horseradish peroxidase, alkaline phosphatase, .beta.-galactosidase, or acetylcholinesterase), a fluorescent material (e.g., umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine, fluorescein, dansyl chloride, allophycocyanin (APC), or phycoerythrin), a luminescent material (e.g., a lanthanide or chelate thereof), a bioluminescent material (e.g., luciferase, luciferin, or aequorin), or a radionuclide (e.g., .sup.3H, .sup.32P, .sup.33P, .sup.125I, or .sup.35S).

[0272] The processing can also involve the coupling of the peptide (or fusion protein) to a polymer (e.g., a polyalkylene glycol moiety such as a polyethylene glycol moiety). In some embodiments, the polymer is coupled to the peptide at a site on the peptide that is an N terminus. In some embodiments, a peptide can contain one or more internal amino acid insertions that provide an internal polymer conjugation site to which a polymer can be conjugated.

[0273] Pharmaceutical Compositions

[0274] Any of the peptides, fusion proteins and nucleic acids encoding the peptides or fusion proteins described herein can be incorporated into pharmaceutical compositions. Such compositions typically include one or more of the peptides (and/or nucleic acids encoding the peptides) and a pharmaceutically acceptable carrier. As used herein the language "pharmaceutically acceptable carrier" includes solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. One or more peptides can be formulated as a pharmaceutical composition in the form of a syrup, an elixir, a suspension, a powder, a granule, a tablet, a capsule, a lozenge, a troche, an aqueous solution, a cream, an ointment, a lotion, a gel, an emulsion, etc. Supplementary active compounds (e.g., one or more chemotherapeutic agents) can also be incorporated into the compositions. Preferably, the composition comprises two or more (e.g., 2, 3, 4, 5, or 6) of the peptides described herein. The composition may also include an immunogenic peptide other than one disclosed herein, e.g., a peptide from WT1 or a derivative thereof, e.g., as described herein. Other immunogenic peptides include, but are not limited to, an immunogenic peptide from MUC1, an immunogenic peptide from gp100, an immunogenic peptide from TRP-2, an immunogenic peptide from MAGI, an immunogenic peptide from NY-ESO1, an immunogenic peptide from HER-2; and an immunogenic peptide from AIM2.

[0275] A pharmaceutical composition is generally formulated to be compatible with its intended route of administration. Examples of routes of administration include oral, rectal, and parenteral, e.g., intravenous, intramuscular, intradermal, subcutaneous, inhalation, transdermal, or transmucosal. Solutions or suspensions used for parenteral application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The compositions can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0276] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL.TM. (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the pharmaceutical composition must be sterile and should be fluid to the extent that easy syringability exists. It should be stable under the conditions of manufacture and storage and must be preserved against any contamination by microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of contamination by microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be desirable to include isotonic agents, for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be facilitated by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

[0277] Sterile injectable solutions can be prepared by incorporating one or more of the peptides (or one or more the nucleic acids encoding the peptides) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the peptide(s) (or fusion proteins or nucleic acid(s) encoding the peptide(s)) into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation can include vacuum drying or freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0278] Oral compositions generally include an inert diluent or an edible carrier. For the purpose of oral therapeutic administration, the one or more peptides (or fusion proteins) can be incorporated with excipients and used in the form of tablets, troches, or capsules, e.g., gelatin capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

[0279] The powders and tablets can contain from 1% to 95% (w/w) of an individual peptide or a mixture of two or more peptides. In certain embodiments, the peptide can range from about 5% to 70% (w/w). Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term "preparation" is intended to include the formulation of the peptide (or nucleic acid) with encapsulating material as a carrier providing a capsule in which the peptide with or without other carriers, is surrounded by a carrier, which is thus in association with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0280] Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizers, and thickening agents as desired. Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents.

[0281] For administration by inhalation, the peptides (or fusion proteins or nucleic acids) can be delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.

[0282] Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the peptides (or fusion proteins or nucleic acids) can be formulated into ointments, salves, gels, or creams as generally known in the art.

[0283] The peptides (or fusion proteins or nucleic acids) can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

[0284] In one embodiment, the peptides (or fusion proteins or nucleic acids) can be prepared with carriers that will protect the peptides (fusion protein or nucleic acid) against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to, e.g., APCs with monoclonal antibodies to APC-specific antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811.

[0285] It can be advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form, as used herein, refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of the peptides (or fusion protein or nucleic acids) calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Dosage units can also be accompanied by instructions for use.

[0286] The nucleic acid molecules encoding the peptides (or fusion protein) can be inserted into vectors and used as gene therapy vectors (as described above). Gene therapy vectors can be delivered to a subject by, for example, intravenous injection, local administration (see, e.g., U.S. Pat. No. 5,328,470) or by stereotactic injection (see, e.g., Chen, et al. (1994) Proc. Natl. Acad. Sci. USA 91: 3054-3057). The pharmaceutical preparation of the gene therapy vector can include the gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. Alternatively, where the complete gene delivery vector can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical preparation can include one or more cells that produce the gene delivery system (see below under "Ex Vivo Methods").

[0287] Additional examples of gene delivery vehicles include, but are not limited to, liposomes, biocompatible polymers, including natural polymers and synthetic polymers; lipoproteins; polypeptides; polysaccharides; lipopolysaccharides; artificial viral envelopes; metal particles; bacteria; viruses such as baculovirus, adenovirus, and retrovirus; bacteriophage; cosmids;

[0288] plasmids; fungal vectors and other recombination vehicles typically used in the art which have been described for expression in a variety of eukaryotic and prokaryotic hosts, and may be used for gene therapy as well as for simple protein expression.

[0289] Examples of viral vectors include retroviral vectors, adenovirus vectors, adeno-associated virus vectors, alphavirus vectors and the like. Liposomes that comprise a targeting moiety such as an antibody or fragment thereof can also be used to prepare pharmaceutical compositions of nucleic acids for delivery to a subject.

[0290] Any of the pharmaceutical compositions described herein can be included in a container, pack, or dispenser together with instructions for administration as described below.

[0291] MHC Molecule Multimer Compositions and Methods for Using the Compositions

[0292] The disclosure also features compositions comprising: (i) one or more of any of the peptides described above and (ii) a major histocompatibility complex (MHC) molecule multimer. The multimer contains two or more (e.g., three, four, five, six, seven, eight, nine, or 10 or more) entire MHC molecules or peptide-binding regions of an MHC molecule. The one or more peptides can be associated with (e.g., covalently or non-covalently bound to) the MHC molecule multimer.

[0293] An MHC molecule of the multimer can be an MHC class I molecule (e.g., an HLA-A molecule such as an HLA-A2 or an HLA-A24 molecule) or an MHC class II molecule. The MHC molecule can be a mammalian (e.g., a rodent, a non-human primate, a human, or any other mammal described herein) MHC molecule.

[0294] The two or more MHC molecules (or the peptide-binding regions of the MHC molecules) in the multimer can be from the same MHC molecule or from different MHC molecules. For example, an MHC molecule multimer can contain five MHC molecules, three of which are the same MHC molecules and two of which are different from the first three. In another example, each MHC molecule of the multimer is different. At least one of the MHC molecules can bind to at least one of the peptides.

[0295] In some embodiments, the above compositions can contain at least two (e.g., two, three, four, five, six, seven, eight, nine, 10, 11, or 15 or more) of any of the peptides described herein. The composition may also include an immunogenic peptide other than one disclosed herein, e.g., a peptide from WT1 or a derivative thereof. Other immunogenic peptides include, but are not limited to, an immunogenic peptide from MUC1, an immunogenic peptide from gp100, an immunogenic peptide from TRP-2, an immunogenic peptide from MAGI, an immunogenic peptide from NY-ESO1, an immunogenic peptide from HER-2; and an immunogenic peptide from AIM2.

[0296] The compositions can also be associated with a detectable label. For example, one or more of the MHC molecules of the multimer can be covalently or non-covalently bound to a detectable label. Suitable detectable labels (e.g., enzymes, fluorescent materials, luminescent materials, bioluminescent materials, or radionuclides) as well as methods for joining detectable labels to a peptide or an MHC molecule are described above.

[0297] An MHC multimer composition can be generated using a peptide described above as follows: a peptide that binds to an HLA molecule is refolded in the presence of the corresponding HLA heavy chain and .beta..sub.2-microglobulin to generate a trimolecular complex. The complex is then biotinylated at the carboxyl terminal end of the heavy chain at a site that was previously engineered into the heavy chain. Multimer formation is then induced by the addition of streptavidin.

[0298] As T cell receptors are capable of recognizing a specific peptide-MHC complex on a target cell among a wide variety of other peptide-MHC complexes, the MHC multimer compositions described herein can be used to, e.g., detect antigen-specific T cells in a population of unrelated T cells (see below). For such assays, the multimers will generally be detectably labeled (see above).

[0299] For example, a multimeric MHC molecule/peptide complex can be used in an assay to assess peripheral blood mononuclear cells for the presence of antigen-specific CTL following exposure to an immunogen. The MHC multimer complex can be used to directly visualize antigen-specific CTL (see, e.g., Ogg et al., Science 279:2103-2106, 1998; and Altman et al., Science 174:94-96, 1996) and determine the frequency of the antigen-specific CTL population in a sample of peripheral blood mononuclear cells. In one example, a detectably-labeled streptavidin used to multimerize the MHC multimer can be used to label T cells that bind to the MHC molecule/peptide complexes of the multimer. To do this, cells treated with the multimer are exposed, e.g., to a label (e.g., a fluorophore conjugated to biotin). The cells can then be readily isolated or detected, e.g., using flow cytometry.

[0300] Applications

[0301] The peptides, fusion proteins (and pharmaceutical compositions thereof), MHC multimer containing compositions, kits, and articles of manufacture described herein can be used in a variety of methods. For example, the peptides described herein can be used to: (i) induce an immune response in a subject (e.g., a subject with a cancer); (ii) activate a T cell in culture (e.g., a central memory T cell and/or effector memory T cell); and/or (iii) treat or even prevent a cancer. Cancers include, e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, blood cell cancers, e.g., plasma cell cancers such as multiple myeloma and leaukemia such as AML or CML. The peptides described herein can also be used to treat precancerous conditions such as smoldering multiple myeloma. As described above, the MHC multimer containing compositions can be used to, e.g., detect antigen-specific T cells in a population of unrelated T cells.

[0302] While the utility of the peptides (or pharmaceutical compositions thereof), MHC multimer containing compositions, kits, or articles of manufacture is in no way limited to any of the particular embodiments described herein, exemplary methods in which these reagents can be used are provided below.

[0303] Methods for Inducing an Immune Response

[0304] The disclosure also features a variety of methods for inducing an immune response in a subject. The methods for inducing an immune response in a subject can include the step of administering to a subject one or more of any of peptides described herein or any of the pharmaceutical compositions described herein. The immune response can be a CD8.sup.+ T cell, a CD4.sup.+ T cell, a cytotoxic T lymphocyte (CTL), a T.sub.H1 response, a T.sub.H2 response, or a combination of both types of responses.

[0305] Any of the above methods can also be, e.g., methods for treating or preventing (prophylaxis against) a cancer (e.g., plasma cell disorder such as multiple myeloma or Waldenstrom's macroglobulinemia, or any other cancer expressing XBP1, CD138, or CS1) in a subject. When the terms "prevent," "preventing," or "prevention" are used herein in connection with a given treatment for a given condition, they mean that the treated subject does not develop a clinically observable level of the condition at all (e.g., the subject does not exhibit one or more symptoms of the condition or, in the case of a cancer, the subject does not develop a detectable level of the cancer),

[0306] As used herein, the term "treat" "treatment," or "treating" a subject having a disorder, e.g., cancer, are used in connection with a given treatment for a given disorder, wherein at least one symptom of the disorder is cured, healed, alleviated, relieved, altered, remedied, ameliorated, or improved. Treating includes administering an amount of a composition effective to alleviate, relieve, alter, remedy, ameliorate, improve or affect the disorder or the symptoms of the disorder. The treatment may inhibit deterioration or worsening of a symptom of a disorder or may cause the condition to develop more slowly and/or to a lesser degree (e.g., fewer symptoms or lower numbers of cancer cells in the subject) in the subject than it would have absent the treatment. For example, a treatment will be said to have "treated" the condition if it is given during the condition, e.g., during an early diagnosis of a cancer (e.g., the detection of a few cancer cells in a sample from the subject) that would have been expected to produce a given manifestation of the condition (an advanced cancer), and results in the subject's experiencing fewer and/or milder symptoms of the condition than otherwise expected. A treatment can "treat" a cancer (e.g., a plasma cell disorder such as multiple myeloma or Waldenstrom's macroglobulinemia) when the subject displays only mild overt symptoms of the cancer.

[0307] In an embodiment, the cancer is a cancer described herein. For example, the cancer can be a cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), and head and neck (e.g, nasopharyngeal cancer).

[0308] Generally, a peptide (or peptides) delivered to the subject will be suspended in a pharmaceutically-acceptable carrier (e.g., physiological saline) and administered orally, rectally, or parenterally, e.g., injected intravenously, subcutaneously, intramuscularly, intrathecally, intraperitoneally, intrarectally, intravaginally, intranasally, intragastrically, intratracheally, or intrapulmonarily (see below).

[0309] Administration can be by periodic injections of a bolus of the pharmaceutical composition or can be uninterrupted or continuous by intravenous or intraperitoneal administration from a reservoir which is external (e.g., an IV bag) or internal (e.g., a bioerodable implant, a bioartificial organ, or a colony of implanted reagent production cells). See, e.g., U.S. Pat. Nos. 4,407,957, 5,798,113 and 5,800,828, each incorporated herein by reference in their entirety.

[0310] In general, the dosage of a peptide or a nucleic acid required depends on the choice of the route of administration; the nature of the formulation; the nature or severity of the subject's illness; the immune status of the subject; the subject's size, weight, surface area, age, and sex; other drugs being administered; and the judgment of the attending medical professional.

[0311] Suitable dosages of peptide for inducing an immune response are in the range of 0.000001 to 10 mg of the reagent or antigenic/immunogenic composition per kg of the subject. Wide variations in the needed dosage are to be expected in view of the variety of reagents and the differing efficiencies of various routes of administration. For example, nasal or rectal administration may require higher dosages than administration by intravenous injection. Variations in these dosage levels can be adjusted using standard empirical routines for optimization as is well understood in the art. Administrations can be single or multiple (e.g., 2-, 3-, 4-, 6-, 8-, 10-, 20-, 50-, 100-, 150-, or more fold). For example, a peptide or peptides can be administered as an initial immunization and then administered one or more times subsequently as a booster immunization.

[0312] In order to optimize therapeutic efficacy (e.g., the efficacy of the one or more peptides or the nucleic acids encoding the peptides to induce an immune response in a subject), compositions containing the peptides or nucleic acids can be first administered at different dosing regimens. The unit dose and regimen depend on factors that include, e.g., the species of mammal, its immune status, the body weight of the mammal.

[0313] The frequency of dosing for a pharmaceutical composition (e.g., a pharmaceutical composition described herein) is within the skills and clinical judgement of medical practitioners (e.g., doctors or nurses). Typically, the administration regime is established by clinical trials which may establish optimal administration parameters. However, the practitioner may vary such administration regimes according to the subject's age, health, weight, sex and medical status.

[0314] In some embodiments, a pharmaceutical composition can be administered to a subject at least two (e.g., three, four, five, six, seven, eight, nine, 10, 11, 12, 15, or 20 or more) times. For example, a pharmaceutical composition can be administered to a subject once a month for three months; once a week for a month; every other week, once a year for three years, once a year for five years; once every five years; once every ten years; or once every three years for a lifetime.

[0315] In some embodiments, the reagent can be administered with an immune modulator such as a Toll Receptor ligand or an adjuvant (see below).

[0316] As defined herein, a "therapeutically effective amount" of a peptide or a nucleic acid encoding a peptide is an amount of the peptide or nucleic acid that is capable of producing an immune response in a treated subject. A therapeutically effective amount of a peptide (i.e., an effective dosage) includes milligram, microgram, nanogram, or picogram amounts of the reagent per kilogram of subject or sample weight (e.g., about 1 nanogram per kilogram to about 500 micrograms per kilogram, about 1 microgram per kilogram to about 500 milligrams per kilogram, about 100 micrograms per kilogram to about 5 milligrams per kilogram, or about 1 microgram per kilogram to about 50 micrograms per kilogram). A therapeutically effective amount of a nucleic acid also includes microgram, nanogram, or picogram amounts of the reagent per kilogram of subject or sample weight (e.g., about 1 nanogram per kilogram to about 500 micrograms per kilogram, about 1 microgram per kilogram to about 500 micrograms per kilogram, about 100 micrograms per kilogram to about 500 micrograms per kilogram, or about 1 microgram per kilogram to about 50 micrograms per kilogram).

[0317] As defined herein, a "prophylactically effective amount" of a peptide or nucleic acid encoding a peptide is an amount of the peptide or nucleic acid that is capable of producing an immune response against a cancer cell (e.g., a breast cancer cell, a colon cancer cell, a pancreatic cancer cell, a prostate cancer cell, a multiple myeloma cell or Waldenstrom's macroglobulinemia cell) in a treated subject, which immune response is capable of preventing the development of a cancer in a subject or is able to substantially reduce the chance of a subject developing or continue developing a cancer (see above). A prophylactically effective amount of a peptide (i.e., an effective dosage) includes milligram, microgram, nanogram, or picogram amounts of the reagent per kilogram of subject or sample weight (e.g., about 1 nanogram per kilogram to about 500 micrograms per kilogram, about 1 microgram per kilogram to about 500 milligrams per kilogram, about 100 micrograms per kilogram to about 5 milligrams per kilogram, or about 1 microgram per kilogram to about 50 micrograms per kilogram). A prophylactically effective amount of a nucleic acid also includes microgram, nanogram, or picogram amounts of the reagent per kilogram of subject or sample weight (e.g., about 1 nanogram per kilogram to about 500 micrograms per kilogram, about 1 microgram per kilogram to about 500 micrograms per kilogram, about 100 micrograms per kilogram to about 500 micrograms per kilogram, or about 1 microgram per kilogram to about 50 micrograms per kilogram).

[0318] The subject can be any animal capable of an immune response to an antigen such as, but not limited to, a mammal, e.g., a human (e.g., a human patient) or a non-human primate (e.g., chimpanzee, baboon, or monkey), mouse, rat, rabbit, guinea pig, gerbil, hamster, horse, a type of livestock (e.g., cow, pig, sheep, or goat), a dog, cat, or a whale. The subject can be one having, suspected of having, or at risk of developing a cancer such as multiple myeloma, Waldenstrom's macroglobulinemia, or any other type of cancer that expresses XBP1, CD138, or CS-1 (e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, leukemia, e.g., AML or CML). The subject can be one in remission from the cancer, e.g., the breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, multiple myeloma or Waldenstrom's macroglobulinemia.

[0319] The methods can also include the step of, prior to administering the one or more peptides (or nucleic acids) to the subject, determining whether one or more cancer cells of the subject's cancer (e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreactic cancer, prostate cancer, leukemia or a plasma cell disorder such as multiple myeloma or Waldenstrom's macroglobulinemia) express XBP1, CD138, or CS-1. Expression of these proteins includes both mRNA and protein expression. Methods for detecting protein and mRNA expression in a cell are known in the art and include, e.g., enzyme-linked immunosorbent assay (ELISA), western and dot-blotting techniques, or immunohistochemistry techniques for detecting protein and reverse transcription-polymerase chain reaction (RT-PCR) or northern-blotting techniques for detecting mRNA. (See Sambrook et al., supra).

[0320] The peptides or composition may be used in combination with other known therapies. Administered "in combination", as used herein, means that two (or more) different treatments are delivered to the subject during the course of the subject's affliction with the disorder, e.g., the two or more treatments are delivered after the subject has been diagnosed with the disorder and before the disorder has been cured or eliminated or treatment has ceased for other reasons. In some embodiments, the delivery of one treatment is still occurring when the delivery of the second begins, so that there is overlap in terms of administration. This is sometimes referred to herein as "simultaneous" or "concurrent delivery". In other embodiments, the delivery of one treatment ends before the delivery of the other treatment begins. In some embodiments of either case, the treatment is more effective because of combined administration. For example, the second treatment is more effective, e.g., an equivalent effect is seen with less of the second treatment, or the second treatment reduces symptoms to a greater extent, than would be seen if the second treatment were administered in the absence of the first treatment, or the analogous situation is seen with the first treatment. In some embodiments, delivery is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one treatment delivered in the absence of the other. The effect of the two treatments can be partially additive, wholly additive, or greater than additive. The delivery can be such that an effect of the first treatment delivered is still detectable when the second is delivered.

[0321] The additional treatment can be, e.g., surgery, one or more chemotherapeutic agents, one or more forms of ionizing radiation, and/or one or more immunomodulatory agents.

[0322] The one or more forms of ionizing radiation can be gamma-irradiation, X-irradiation, or beta-irradiation.

[0323] Exemplary classes of chemotherapeutic agents include, e.g., the following:

[0324] alkylating agents (including, without limitation, nitrogen mustards, ethylenimine derivatives, alkyl sulfonates, nitrosoureas and triazenes): uracil mustard (Aminouracil Mustard.RTM., Chlorethaminacil.RTM., Demethyldopan.RTM., Desmethyldopan.RTM., Haemanthamine.RTM., Nordopan.RTM., Uracil nitrogen Mustard.RTM., Uracillost.RTM., Uracilmostaza.RTM., Uramustin.RTM., Uramustine.RTM.), chlormethine (Mustargen.RTM.), cyclophosphamide (Cytoxan.RTM., Neosar.RTM., Clafen.RTM., Endoxan.RTM., Procytox.RTM., Revimmune.TM.), ifosfamide (Mitoxana.RTM.), melphalan (Alkeran.RTM.), Chlorambucil (Leukeran.RTM.), pipobroman (Amedel.RTM., Vercyte.RTM.), triethylenemelamine (Hemel.RTM., Hexalen.RTM., Hexastat.RTM.), triethylenethiophosphoramine, Temozolomide (Temodar.RTM.), thiotepa (Thioplex.RTM.), busulfan (Busilvex.RTM., Myleran.RTM.), carmustine (BiCNU.RTM.), lomustine (CeeNU.RTM.), streptozocin (Zanosar.RTM.), and Dacarbazine (DTIC-Dome.RTM.).

[0325] anti-EGFR antibodies (e.g., cetuximab (Erbitux.RTM.) and panitumumab (Vectibix.RTM.).

[0326] anti-HER-2 antibodies (e.g., trastuzumab (Herceptin.RTM.).

[0327] antimetabolites (including, without limitation, folic acid antagonists (also referred to herein as antifolates), pyrimidine analogs, purine analogs and adenosine deaminase inhibitors): methotrexate (Rheumatrex.RTM., Trexall.RTM.), 5-fluorouracil (Adrucil.RTM., Efudex.RTM., Fluoroplex.RTM.), floxuridine (FUDF.RTM.), cytarabine (Cytosar-U.RTM., Tarabine PFS), 6-mercaptopurine (Puri-Nethol.RTM.)), 6-thioguanine (Thioguanine Tabloid.RTM.), fludarabine phosphate (Fludara.RTM.), pentostatin (Nipent.RTM.), pemetrexed (Alimta.RTM.), raltitrexed (Tomudex.RTM.), cladribine (Leustatin.RTM.), clofarabine (Clofarex.RTM., Clolar.RTM.), mercaptopurine (Puri-Nethol.RTM.), capecitabine (Xeloda.RTM.), nelarabine (Arranon.RTM.), azacitidine (Vidaza.RTM.) and gemcitabine (Gemzar.RTM.). Preferred antimetabolites include, e.g., 5-fluorouracil (Adrucil.RTM., Efudex.RTM., Fluoroplex.RTM.), floxuridine (FUDF.RTM.), capecitabine (Xeloda.RTM.), pemetrexed (Alimta.RTM.), raltitrexed (Tomudex.RTM.) and gemcitabine (Gemzar.RTM.).

[0328] vinca alkaloids: vinblastine (Velban.RTM., Velsar.RTM.), vincristine (Vincasar.RTM., Oncovin.RTM.), vindesine (Eldisine.RTM.), vinorelbine (Navelbine.RTM.).

[0329] platinum-based agents: carboplatin (Paraplat.RTM., Paraplatin.RTM.), cisplatin (Platinol.RTM.), oxaliplatin (Eloxatin.RTM.).

[0330] anthracyclines: daunorubicin (Cerubidine.RTM., Rubidomycin.RTM.), doxorubicin (Adriamycin.RTM.), epirubicin (Ellence.RTM.), idarubicin (Idamycin.RTM.), mitoxantrone (Novantrone.RTM.), valrubicin (Valstar.RTM.). Preferred anthracyclines include daunorubicin (Cerubidine.RTM., Rubidomycin.RTM.) and doxorubicin (Adriamycin.RTM.).

[0331] topoisomerase inhibitors: topotecan (Hycamtin.RTM.), irinotecan (Camptosar.RTM.), etoposide (Toposar.RTM., VePesid.RTM.), teniposide (Vumon.RTM.), lamellarin D, SN-38, camptothecin.

[0332] taxanes: paclitaxel (Taxol.RTM.), docetaxel (Taxotere.RTM.), larotaxel, cabazitaxel.

[0333] epothilones: ixabepilone, epothilone B, epothilone D, BMS310705, dehydelone, ZK-Epothilone (ZK-EPO).

[0334] poly ADP-ribose polymerase (PARP) inhibitors: (e.g., BSI 201, Olaparib (AZD-2281), ABT-888, AG014699, CEP 9722, MK 4827, KU-0059436 (AZD2281), LT-673, 3-aminobenzamide).

[0335] antibiotics: actinomycin (Cosmegen.RTM.), bleomycin (Blenoxane.RTM.), hydroxyurea (Droxia.RTM., Hydrea.RTM.), mitomycin (Mitozytrex.RTM., Mutamycin.RTM.).

[0336] immunomodulators: lenalidomide (Revlimid.RTM.), thalidomide (Thalomid.RTM.).

[0337] immune cell antibodies: alemtuzamab (Campath.RTM.), gemtuzumab (Myelotarg.RTM.), rituximab (Rituxan.RTM.), tositumomab (Bexxar.RTM.).

[0338] interferons (e.g., IFN-alpha (Alferon.RTM., Roferon-A.RTM., Intron.RTM.-A) or IFN-gamma (Actimmune.RTM.)).

[0339] interleukins: IL-1, IL-2 (Proleukin.RTM.), IL-24, IL-6 (Sigosix.RTM.), IL-12.

[0340] HSP90 inhibitors (e.g., geldanamycin or any of its derivatives). In certain embodiments, the HSP90 inhibitor is selected from geldanamycin, 17-alkylamino-17-desmethoxygeldanamycin ("17-AAG") or 17-(2-dimethylaminoethyl)amino-17-desmethoxygeldanamycin ("17-DMAG").

[0341] angiogenesis inhibitors which include, without limitation A6 (Angstrom Pharmacueticals), ABT-510 (Abbott Laboratories), ABT-627 (Atrasentan) (Abbott Laboratories/Xinlay), ABT-869 (Abbott Laboratories), Actimid (CC4047, Pomalidomide) (Celgene Corporation), AdGVPEDF.11D (GenVec), ADH-1 (Exherin) (Adherex Technologies), AEE788 (Novartis), AG-013736 (Axitinib) (Pfizer), AG3340 (Prinomastat) (Agouron Pharmaceuticals), AGX1053 (AngioGenex), AGX51 (AngioGenex), ALN-VSP (ALN-VSP 02) (Alnylam Pharmaceuticals), AMG 386 (Amgen), AMG706 (Amgen), Apatinib (YN968D1) (Jiangsu Hengrui Medicine), AP23573 (Ridaforolimus/MK8669) (Ariad Pharmaceuticals), AQ4N (Novavea), ARQ 197 (ArQule), ASA404 (Novartis/Antisoma), Atiprimod (Callisto Pharmaceuticals), ATN-161 (Attenuon), AV-412 (Aveo Pharmaceuticals), AV-951 (Aveo Pharmaceuticals), Avastin (Bevacizumab) (Genentech), AZD2171 (Cediranib/Recentin) (AstraZeneca), BAY 57-9352 (Telatinib) (Bayer), BEZ235 (Novartis), BIBF1120 (Boehringer Ingelheim Pharmaceuticals), BIBW 2992 (Boehringer Ingelheim Pharmaceuticals), BMS-275291 (Bristol-Myers Squibb), BMS-582664 (Brivanib) (Bristol-Myers Squibb), BMS-690514 (Bristol-Myers Squibb), Calcitriol, CCI-779 (Torisel) (Wyeth), CDP-791 (ImClone Systems), Ceflatonin (Homoharringtonine/HHT) (ChemGenex Therapeutics), Celebrex (Celecoxib) (Pfizer), CEP-7055 (Cephalon/Sanofi), CHIR-265 (Chiron Corporation), NGR-TNF, COL-3 (Metastat) (Collagenex Pharaceuticals), Combretastatin (Oxigene), CP-751,871 (Figitumumab) (Pfizer), CP-547,632 (Pfizer), CS-7017 (Daiichi Sankyo Pharma), CT-322 (Angiocept) (Adnexus), Curcumin, Dalteparin (Fragmin) (Pfizer), Disulfiram (Antabuse), E7820 (Eisai Limited), E7080 (Eisai Limited), EMD 121974 (Cilengitide) (EMD Pharmaceuticals), ENMD-1198 (EntreMed), ENMD-2076 (EntreMed), Endostar (Simcere), Erbitux (ImClone/Bristol-Myers Squibb), EZN-2208 (Enzon Pharmaceuticals), EZN-2968 (Enzon Pharmaceuticals), GC1008 (Genzyme), Genistein, GSK1363089 (Foretinib) (GlaxoSmithKline), GW786034 (Pazopanib) (GlaxoSmithKline), GT-111 (Vascular Biogenics Ltd.), IMC-1121B (Ramucirumab) (ImClone Systems), IMC-18F1 (ImClone Systems), IMC-3G3 (ImClone LLC), INCB007839 (Incyte Corporation), INGN 241 (Introgen Therapeutics), Iressa (ZD1839/Gefitinib), LBH589 (Faridak/Panobinostst) (Novartis), Lucentis (Ranibizumab) (Genentech/Novartis), LY317615 (Enzastaurin) (Eli Lilly and Company), Macugen (Pegaptanib) (Pfizer), MEDI522 (Abegrin) (MedImmune), MLN518 (Tandutinib) (Millennium), Neovastat (AE941/Benefin) (Aeterna Zentaris), Nexavar (Bayer/Onyx), NM-3 (Genzyme Corporation), Noscapine (Cougar Biotechnology), NPI-2358 (Nereus Pharmaceuticals), OSI-930 (OSI), Palomid 529 (Paloma Pharmaceuticals, Inc.), Panzem Capsules (2ME2) (EntreMed), Panzem NCD (2ME2) (EntreMed), PF-02341066 (Pfizer), PF-04554878 (Pfizer), PI-88 (Progen Industries/Medigen Biotechnology), PKC412 (Novartis), Polyphenon E (Green Tea Extract) (Polypheno E International, Inc), PPI-2458 (Praecis Pharmaceuticals), PTC299 (PTC Therapeutics), PTK787 (Vatalanib) (Novartis), PXD101 (Belinostat) (CuraGen Corporation), RAD001 (Everolimus) (Novartis), RAF265 (Novartis), Regorafenib (BAY73-4506) (Bayer), Revlimid (Celgene), Retaane (Alcon Research), SN38 (Liposomal) (Neopharm), SNS-032 (BMS-387032) (Sunesis), SOM230 (Pasireotide) (Novartis), Squalamine (Genaera), Suramin, Sutent (Pfizer), Tarceva (Genentech), TB-403 (Thrombogenics), Tempostatin (Collard Biopharmaceuticals), Tetrathiomolybdate (Sigma-Aldrich), TG100801 (TargeGen), Thalidomide (Celgene Corporation), Tinzaparin Sodium, TKI258 (Novartis), TRC093 (Tracon Pharmaceuticals Inc.), VEGF Trap (Aflibercept) (Regeneron Pharmaceuticals), VEGF Trap-Eye (Regeneron Pharmaceuticals), Veglin (VasGene Therapeutics), Bortezomib (Millennium), XL184 (Exelixis), XL647 (Exelixis), XL784 (Exelixis), XL820 (Exelixis), XL999 (Exelixis), ZD6474 (AstraZeneca), Vorinostat (Merck), and ZSTK474.

[0342] anti-androgens which include, without limitation nilutamide (Nilandron.RTM.) and bicalutamide (Caxodex.RTM.).

[0343] antiestrogens which include, without limitation tamoxifen (Nolvadex.RTM.), toremifene (Fareston.RTM.), letrozole (Femara.RTM.), testolactone (Teslac.RTM.), anastrozole (Arimidex.RTM.), bicalutamide (Casodex.RTM.), exemestane (Aromasin.RTM.), flutamide (Eulexin.RTM.), fulvestrant (Faslodex.RTM.), raloxifene (Evista.RTM., Keoxifene.RTM.) and raloxifene hydrochloride.

[0344] anti-hypercalcaemia agents which include without limitation gallium (III) nitrate hydrate (Ganite.RTM.) and pamidronate disodium (Aredia.RTM.).

[0345] apoptosis inducers which include without limitation ethanol, 2-[[3-(2,3-dichlorophenoxy)propyl]amino]-(9Cl), gambogic acid, embelin and arsenic trioxide (Trisenox.RTM.).

[0346] Aurora kinase inhibitors which include without limitation binucleine 2.

[0347] Bruton's tyrosine kinase inhibitors which include without limitation terreic acid.

[0348] calcineurin inhibitors which include without limitation cypermethrin, deltamethrin, fenvalerate and tyrphostin 8.

[0349] CaM kinase II inhibitors which include without limitation 5-Isoquinolinesulfonic acid, 4-[{2S)-2-[(5-isoquinolinylsulfonyl)methylamino]-3-oxo-3-14-phenyl-1-pipe- razinyl)propyl]phenyl ester and benzenesulfonamide.

[0350] CD45 tyrosine phosphatase inhibitors which include without limitation phosphonic acid.

[0351] CDC25 phosphatase inhibitors which include without limitation 1,4-naphthalene dione, 2,3-bis[(2-hydroxyethyl)thio]-(9Cl).

[0352] CHK kinase inhibitors which include without limitation debromohymenialdisine.

[0353] cyclooxygenase inhibitors which include without limitation 1H-indole-3-acetamide, 1-(4-chlorobenzoyl)-5-methoxy-2-methyl-N-(2-phenylethyl)-(9Cl), 5-alkyl substituted 2-arylaminophenylacetic acid and its derivatives (e.g., celecoxib (Celebrex.RTM.), rofecoxib (Vioxx.RTM.), etoricoxib (Arcoxia.RTM.), lumiracoxib (Prexige.RTM.), valdecoxib (Bextra.RTM.) or 5-alkyl-2-arylaminophenylacetic acid).

[0354] cRAF kinase inhibitors which include without limitation 3-(3,5-dibromo-4-hydroxybenzylidene)-5-iodo-1,3-dihydroindol-2-one and benzamide, 3-(dimethylamino)-N-[3-[(4-hydroxybenzoyl)amino]-4-methylphenyl]-(9Cl).

[0355] cyclin dependent kinase inhibitors which include without limitation olomoucine and its derivatives, purvalanol B, roascovitine (Seliciclib.RTM.), indirubin, kenpaullone, purvalanol A and indirubin-3'-monooxime.

[0356] cysteine protease inhibitors which include without limitation 4-morpholinecarboxamide, N-[(1S)-3-fluoro-2-oxo-1-(2-phenylethyl)propyl]amino]-2-oxo-1-(phenylmeth- yl)ethyl]-(9Cl).

[0357] DNA intercalators which include without limitation plicamycin (Mithracin.RTM.) and daptomycin (Cubicin.RTM.).

[0358] DNA strand breakers which include without limitation bleomycin (Blenoxane.RTM.).

[0359] E3 ligase inhibitors which include without limitation N-((3,3,3-trifluoro-2-trifluoromethyl)propionyl)sulfanilamide.

[0360] EGF Pathway Inhibitors which include, without limitation tyrphostin 46, EKB-569, erlotinib (Tarceva.RTM.), gefitinib (Iressa.RTM.), lapatinib (Tykerb.RTM.) and those compounds that are generically and specifically disclosed in WO 97/02266, EP 0 564 409, WO 99/03854, EP 0 520 722, EP 0 566 226, EP 0 787 722, EP 0 837 063, U.S. Pat. No. 5,747,498, WO 98/10767, WO 97/30034, WO 97/49688, WO 97/38983 and WO 96/33980.

[0361] farnesyltransferase inhibitors which include without limitation A-hydroxyfarnesylphosphonic acid, butanoic acid, 2-[(2S)-2-[[(2S,3S)-2-[[(2R)-2-amino-3-mercaptopropyl]amino]-3-methylpent- yl]oxy]-1-oxo-3-phenylpropyl]amino]-4-(methylsulfonyl)-1-methylethylester (2S)-(9Cl), and manumycin A.

[0362] Flk-1 kinase inhibitors which include without limitation 2-propenamide, 2-cyano-3-[4-hydroxy-3,5-bis(1-methylethyl)phenyl]-N-(3-phenylpropyl)-(2E- )-(9Cl).

[0363] glycogen synthase kinase-3 (GSK3) inhibitors which include without limitation indirubin-3'-monooxime.

[0364] Heat Shock Protein 90 (Hsp90) chaperone modulators which include without limitation AUY922, STA-9090, ATI13387, MCP-3100, IPI-504, IPI-493, SNX-5422, Debio0932, HSP990, DS-2248, PU-H71, 17-DMAG (Alvespimycin), and XL888.

[0365] histone deacetylase (HDAC) inhibitors which include without limitation suberoylanilide hydroxamic acid (SAHA), [4-(2-amino-phenylcarbamoyl)-benzyl]-carbamic acid pyridine-3-ylmethylester and its derivatives, butyric acid, pyroxamide, trichostatin A, oxamflatin, apicidin, depsipeptide, depudecin, trapoxin and compounds disclosed in WO 02/22577.

[0366] I-kappa B-alpha kinase inhibitors (IKK) which include without limitation 2-propenenitrile, 3-[(4-methylphenyl)sulfonyl]-(2E)-(9Cl).

[0367] imidazotetrazinones which include without limitation temozolomide (Methazolastone.RTM., Temodar.RTM. and its derivatives (e.g., as disclosed generically and specifically in U.S. Pat. No. 5,260,291) and Mitozolomide.

[0368] Insulin like growth factor pathway inhibitors such as IGF inhibitors or IGF receptor (IGFR1 or IGFR2) inhibitors include without limitation, small molecule inhibitors, e.g., OSI-906; anti-IGF antibodies or anti-IGFR antibodies, e.g., AVE-1642, MK-0646, IMC-A12 (cixutumab), R1507, CP-751,871 (Figitumumab).

[0369] insulin tyrosine kinase inhibitors which include without limitation hydroxyl-2-naphthalenylmethylphosphonic acid.

[0370] c-Jun-N-terminal kinase (JNK) inhibitors which include without limitation pyrazoleanthrone and epigallocatechin gallate.

[0371] mitogen-activated protein kinase (MAP) inhibitors which include without limitation benzenesulfonamide, N-[2-[[[3-(4-chlorophenyl)-2-propenyl]methyl]amino]methyl]phenyl]-N-(2-hy- droxyethyl)-4-methoxy-(9Cl).

[0372] MDM2 inhibitors which include without limitation trans-4-iodo, 4'-boranyl-chalcone.

[0373] MEK inhibitors which include without limitation butanedinitrile, bis[amino[2-aminophenyl)thio]methylene]-(9Cl).

[0374] MMP inhibitors which include without limitation Actinonin, epigallocatechin gallate, collagen peptidomimetic and non-peptidomimetic inhibitors, tetracycline derivatives marimastat (Marimastat.RTM.), prinomastat, incyclinide (Metastat.RTM.), shark cartilage extract AE-941 (Neovastat.RTM.), Tanomastat, TAA211, MMI270B or AAJ996.

[0375] mTor inhibitors which include without limitation rapamycin (Rapamune.RTM.), and analogs and derivatives thereof, AP23573 (also known as ridaforolimus, deforolimus, or MK-8669), CCI-779 (also known as temsirolimus) (Torisel.RTM.) and SDZ-RAD.

[0376] NGFR tyrosine kinase inhibitors which include without limitation tyrphostin AG 879.

[0377] p38 MAP kinase inhibitors which include without limitation Phenol, 4-[4-(4-fluorophenyl)-5-(4-pyridinyl)-1H-imidazol-2-yl]-(9Cl), and benzamide, 3-(dimethylamino)-N-[3-[(4-hydroxylbenzoyl)amino]-4-methylphenyl]-(9Cl).

[0378] p56 tyrosine kinase inhibitors which include without limitation damnacanthal and tyrphostin 46.

[0379] PDGF pathway inhibitors which include without limitation tyrphostin AG 1296, tyrphostin 9, 1,3-butadiene-1,1,3-tricarbonitrile, 2-amino-4-(1H-indol-5-yl)-(9Cl), imatinib (Gleevec.RTM.) and gefitinib (Iressa.RTM.) and those compounds generically and specifically disclosed in European Patent No.: 0 564 409 and PCT Publication No.: WO 99/03854.

[0380] phosphatidylinositol 3-kinase inhibitors which include without limitation wortmannin, and quercetin dihydrate.

[0381] phosphatase inhibitors which include without limitation cantharidic acid, cantharidin, and L-leucinamide.

[0382] PKC inhibitors which include without limitation 1-H-pyrollo-2,5-dione,3-[1-[3-(dimethylamino)propyl]-1H-indol-3-yl]-4-(1H- -indol-3-yl)-(9Cl), Bisindolylmaleimide IX, Sphinogosine, staurosporine, and Hypericin.

[0383] PKC delta kinase inhibitors which include without limitation rottlerin.polyamine synthesis inhibitors which include without limitation DMFO.

[0384] proteasome inhibitors which include, without limitation aclacinomycin A, gliotoxin and bortezomib (Velcade.RTM.).

[0385] protein phosphatase inhibitors which include without limitation cantharidic acid, cantharidin, L-P-bromotetramisole oxalate, 2(5H)-furanone, 4-hydroxy-5-(hydroxymethyl)-3-(1-oxohexadecyl)-(5R)-(9Cl) and benzylphosphonic acid.

[0386] protein tyrosine kinase inhibitors which include, without limitation tyrphostin Ag 216, tyrphostin Ag 1288, tyrphostin Ag 1295, geldanamycin, genistein and 7H-pyrollo[2,3-d]pyrimidine derivatives;

[0387] PTP1B inhibitors which include without limitation L-leucinamide.

[0388] SRC family tyrosine kinase inhibitors which include without limitation PP1 and PP2.

[0389] Syk tyrosine kinase inhibitors which include without limitation piceatannol.

[0390] Janus (JAK-2 and/or JAK-3) tyrosine kinase inhibitors which include without limitation tyrphostin AG 490 and 2-naphthyl vinyl ketone.

[0391] retinoids which include without limitation isotretinoin (Accutane.RTM., Amnesteem.RTM., Cistane.RTM., Claravis.RTM., Sotret.RTM.) and tretinoin (Aberel.RTM., Aknoten.RTM., Avita.RTM., Renova.RTM., Retin-A.RTM., Retin-A MICRO.RTM., Vesanoid.RTM.).

[0392] RNA polymerase II elongation inhibitors which include without limitation 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole.

[0393] serine/threonine kinase inhibitors which include without limitation 2-aminopurine.

[0394] sterol biosynthesis inhibitors which include without limitation squalene epoxidase and CYP2D6.VEGF pathway inhibitors which include without limitation anti-VEGF antibodies, e.g., bevacizumab, and small molecules, e.g., sunitinib (Sutent.RTM.), sorafinib (Nexavar.RTM.), ZD6474 (also known as vandetanib) (Zactima.TM.), SU6668, CP-547632, AV-951 (tivozanib) and AZD2171 (also known as cediranib) (Recentin.TM.).

[0395] For example, one or more chemotherapeutic agents can be selected from the group consisting of cisplatin, carboplatin, procarbazine, mechlorethamine, cyclophosphamide, camptothecin, adriamycin, ifosfamide, melphalan, chlorambucil, bisulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, mitomycin, etoposide, verampil, podophyllotoxin, tamoxifen, taxol, thalidomide, lenalidomide, a proteosome inhibitor (e.g., bortezomib), an hsp90 inhibitor (e.g., tenespinmycin), transplatinum, 5-flurouracil, vincristin, vinblastin, methotrexate, or an analog of any of the aforementioned. Immunomodulatory agents include, e.g., a variety of chemokines and cytokines such as Interleukin 2 (IL-2), granulocyte/macrophage-colony stimulating factor (GM-CSF), and Interleukin 12 (IL-12).

[0396] In one embodiment, the additional therapy is one or more additional immunogenic peptide, e.g., one or more immunogenic peptide from WT1 or a derivative thereof. Exemplary WT1 immunogenic peptides include, but are not limited to, a WT1 class 1 epitope; a peptide comprising (or consisting of) RMFPNAPYL (SEQ ID NO: 538) (WT1 126-134); a peptide comprising (or consisting of) YMFPNAPYL (SEQ ID NO: 539); a peptide comprising (or consisting of) RSDELVRHHNMHQRNMTKL (SEQ ID NO: 540) (WT1 427-445); a peptide comprising (or consisting of) PGCNKRYFKLSHLQMHSRKHTG (SEQ ID NO: 541) (WT1 331-352); a peptide comprising (or consisting of) SGQARMFPNAPYLPSCLES (SEQ ID NO: 542) (WT1 122-140); and a peptide comprising (or consisting of) SGQAYMFPNAPYLPSCLES (SEQ ID NO: 543). Other WT1 immunogenic peptides are described in U.S. Pat. No. 7,598,221, the contents of which is incorporated herein by reference. Other immunogenic peptides include, but are not limited to, an immunogenic peptide from MUC1, an immunogenic peptide from gp100, an immunogenic peptide from TRP-2, an immunogenic peptide from MAGI, an immunogenic peptide from NY-ESO1, an immunogenic peptide from HER-2; and an immunogenic peptide from AIM2.

[0397] The subject can have, be suspected of having, or be at risk of developing a cancer such as lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreactic cancer, prostate cancer, leukemia, multiple myeloma or Waldenstrom's macroglobulinemia. A subject "suspected of having a cancer" is one having one or more symptoms of a cancer. Symptoms of cancer are well-known to those of skill in the art and generally include, without limitation, pain, weight loss, weakness, excessive fatigue, difficulty eating, loss of appetite, chronic cough, worsening breathlessness, coughing up blood, blood in the urine, blood in stool, nausea, vomiting, abdominal fullness, bloating, fluid in peritoneal cavity, vaginal bleeding, constipation, abdominal distension, perforation of colon, acute peritonitis (infection, fever, pain), pain, vomiting blood, heavy sweating, fever, high blood pressure, anemia, diarrhea, jaundice, dizziness, chills, muscle spasms, difficulty swallowing, and the like. Symptoms of multiple myeloma specifically include, e.g., bone pain (e.g., in the back or ribs), high levels of calcium in the blood, excessive thirst or urination, constipation, nausea, loss of appetite, confusion, weakness or numbness in the legs, weight loss, or repeated infections. Symptoms indicative of Waldenstrom's macroglobulinemia include, e.g., weakness, swollen lymph nodes, severe fatigue, nose bleeds, weight loss, neurological problems.

[0398] As used herein, a subject "at risk of developing a cancer" is a subject that has a predisposition to develop a cancer, i.e., a genetic predisposition to develop cancer such as a mutation in a tumor suppressor gene (e.g., mutation in BRCA1, p53, RB, or APC), has been exposed to conditions, or is presently affected by conditions, that can result in cancer. Thus, a subject can also be one "at risk of developing a cancer" when the subject has been exposed to mutagenic or carcinogenic levels of certain compounds (e.g., carcinogenic compounds in cigarette smoke such as acrolein, 4-aminobiphenyl, aromatic amines, benzene, benz{a}anthracene, benzo{a}pyrene, formaldehyde, hydrazine, Polonium-210 (Radon), urethane, or vinyl chloride). The subject can be "at risk of developing a cancer" when the subject has been exposed to, e.g., large doses of ultraviolet light or X-irradiation, or exposed (e.g., infected) to a tumor-causing/associated virus such as papillomavirus, Epstein-Barr virus, hepatitis B virus, or human T-cell leukemia-lymphoma virus. In addition, a subject can be "at risk of developing a cancer" when the subject suffers from an inflammation (e.g., chronic inflammation). A subject can be at risk of developing multiple myeloma if, e.g., the subject has monoclonal gammopathy of undetermined significance (MGUS). A subject can be at risk of developing any cancer described herein, for example, cancer of the bladder (including accelerated and metastatic bladder cancer), breast (e.g., estrogen receptor positive breast cancer, estrogen receptor negative breast cancer, HER-2 positive breast cancer, HER-2 negative breast cancer, triple negative breast cancer, inflammatory breast cancer), colon (including colorectal cancer), kidney (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver, lung (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchoalveolar carcinoma and large cell carcinoma)), genitourinary tract, e.g., ovary (including fallopian, endometrial and peritoneal cancers), cervix, prostate and testes, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic carcinoma), stomach (e.g., gastroesophageal, upper gastric or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gall bladder, thyroid, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), head and neck, or multiple myeloma.

[0399] Multiple myeloma is a hematological cancer that affects approximately 45,000 people in United States each year. Multiple myeloma is characterized by a multifocal proliferation and clonal expansion of plasma cells within the bone marrow, which can result in skeletal dysfunction, serum monoclonal gammopathy, immunosuppression and end-organ sequelae. Smoldering multiple myeloma (SMM) patients are at a high risk for progression to active multiple myeloma. While treatments including bone marrow transplant and chemotherapy are used to treat patients that develop active multiple myeloma, the prognosis of patients with these stage of the disease is often poor.

[0400] Smoldering multiple myeloma (SMM) is an asymptomatic, plasma cell proliferative disorder characterized by monoclonal plasma cell proliferation in the bone marrow and monoclonal proteins in the blood and/or urine without renal dysfunction, hypercalcemia, bone disease, or anemia. The diagnosis of SMM requires a serum monoclonal (M) protein level >3 g/dL and/or bone marrow clonal plasma cells (BMPC)>10%, and the absence of end-organ damage (i.e., hypercalcemia, renal insufficiency, anemia, or bone lesions [CRAB]). Although asymptomatic, SMM is associated with a high-risk of progression to symptomatic multiple myeloma (MM) or amyloidosis.

[0401] Currently, there is no active treatment for SMM. Instead, a "watchful" waiting approach is taken, with treatment initiated after progression to symptomatic disease. For most patients, progression is indicated by an increase in anemia (hemoglobin lower than 2 g/dL below the lower limit of normal or <10 g/dL) and/or skeletal involvement, including bone lesions and/or diffuse osteoporosis.

[0402] A subject having smoldering multiple myeloma is also a subject at risk for developing multiple myeloma. Smoldering multiple myeloma can be determined by increased levels of monoclonal protein, e.g., in the urine or blood of a subject. In addition, subjects having smoldering multiple myeloma can exhibit increased numbers of myeloma cells in the bone marrow. At present, it has been estimated that SMM accounts for approximately 15% of all newly diagnosed cases of MM. The median time to progression from diagnosis to symptomatic MM ranges from 2 to 3 years, and the annual risk of progression from SMM to symptomatic MM requiring treatment is estimated to be 10%. The risk of progression is contingent on 1) monoclonal protein levels >3 g/dL; 2) BMPC>10%; and 3) the presence of abnormal serum free light chain (FLC) ratio. As shown in Table 1, the median time to progression is notably shorter in patients meeting all 3 of these prognostic criteria (1.9 years) compared to those meeting only 1 of these criteria (10 years) or 2 of these criteria (5.1 years). Similarly, the proportion of patients progressing to symptomatic MM by 5 years is notably higher in those meeting all 3 prognostic criteria (76%) compared to those meeting only 1 or 2 of these criteria (25% and 51%, respectively).

TABLE-US-00004 TABLE 4 SMM Prognosis Number of Median Time to Progression at Number of Risk Patients Progression 5 Years Factors (N) (Years) (%) 1 81 10 25 2 114 5.1 51 3 78 1.9 76 Source: Kyle RA, et al, 2010.

[0403] Recent data have demonstrated the importance of other prognostic criteria for SMM, including the presence of an aberrant phenotype of BMPC defined as a decrease in 1 or 2 of the uninvolved immunoglobulin (Ig) isotypes as well as whether M protein remains stable or progressively worsens over time. In this latter case, referred to as evolving SMM, patients with a progressive increase in serum M protein levels have shorter median time to progression (TTP) compared to those with a stable M protein of 1.3 versus 3.9 years, respectively.

[0404] In some embodiments, the method can also include determining if an immune response occurred in a subject after administering a peptide(s), nucleic acids or composition described herein to the subject. Suitable methods for determining whether an immune response occurred in a subject include use of immunoassays to detect, e.g., the presence of antibodies specific for a peptide in a biological sample from the subject. For example, after the administration of the peptide or composition to the subject, a biological sample (e.g., a blood sample) can be obtained from the subject and tested for the presence of antibodies specific for the peptide(s). An immune response can also be detected by assaying for the presence or amount of activated T cells in a sample. Such assays include, e.g., proliferation assays, limiting dilution assays, cytotoxicity assays (e.g., lymphokine- or .sup.51Cr-release assays)(as described above).

[0405] In some embodiments, the methods can also include the step of determining whether a subject has a cancer. Suitable methods for such a determination depend on the type of cancer to be detected in the subject, but are known in the art. Such methods can be qualitative or quantitative. For example, a medical practitioner can diagnose a subject as having multiple myeloma when the subject exhibits two or more (e.g., three, four, five, or six or more) symptoms of multiple myeloma such as any of those described herein. A subject can also be determined to have multiple myeloma by measuring the blood calcium level, the white or red blood cell count, or the amount of protein in the urine of a subject.

[0406] Ex Vivo Approaches.

[0407] An ex vivo strategy for inducing an immune response in a subject can involve contacting suitable APCs (e.g., dendritic cells, monocytes, or macrophages) obtained from the subject with any of the peptides or compositions described herein. Alternatively, the cells can be transfected with a nucleic acid (e.g., an expression vector) encoding one or more of the peptides and optionally cultured for a period of time and under conditions that permit the expression of the peptides. The transfection method will depend on the type of cell and nucleic acid being transfected into the cell. (See above under "Nucleic Acids and Methods for Producing the Peptides" and also Sambrook et al., supra). Following the contacting or transfection, the cells are then returned to the subject.

[0408] The cells can be any of a wide range of types expressing MHC class I or II molecules. For example, the cells can include bone marrow cells, macrophages, monocytes, dendritic cells, T cells (e.g., T helper cells, CD4.sup.+ cells, CD8.sup.+ cells, or cytotoxic T cells), or B cells.

[0409] Ex vivo methods for stimulating an immune response can include contacting in vitro a T cell (e.g., in a population of lymphocytes obtained from a subject) with an antigen-presenting cell expressing an MHC molecule bound to one of the peptides described herein for an amount of time (and under conditions) that is sufficient to activate the T cell. Following the contacting, the activated T cell(s) are reintroduced into the subject from which the cells were obtained. Methods for generating an APC expressing an MHC molecule bound to one of the peptides described herein are set forth above in this section.

[0410] In some embodiments of any of the ex vivo methods, cells can be obtained from a subject of the same species other than the subject (allogeneic) can be contacted with the reagents (or immunogenic/antigenic compositions) and administered to the subject.

[0411] Methods for Producing an Antibody in a Subject

[0412] Methods of producing an antibody specific for an immunogen (e.g., one or more of any of the peptides described herein) are known in the art and detailed below. For example, antibodies or antibody fragments specific for a peptide described herein can be generated by immunization, e.g., using an animal, or by in vitro methods such as phage display. All or part of a peptide described herein can be used to generate an antibody or antibody fragment.

[0413] A peptide can be used to prepare antibodies by immunizing a suitable subject, (e.g., rabbit, goat, mouse, or other mammal such as a human) with the peptide. An appropriate immunogenic preparation can contain, for example, any of the reagents described herein. The preparation can further include an adjuvant, such as Freund's complete or incomplete adjuvant, alum, RIBI, or similar immunostimulatory agent. Adjuvants also include, e.g., cholera toxin (CT), E. coli heat labile toxin (LT), mutant CT (MCT) (Yamamoto et al. (1997) J. Exp. Med. 185:1203-1210) mutant E. coli heat labile toxin (MLT) (Di Tommaso et al. (1996) Infect. Immunity 64:974-979), a combination of carboxymethylcellulose, polyinosinic-polycytidylic acid, and poly-L-lysine double-stranded RNA (e.g., poly IC-LC, e.g., hiltonol), a water-and-oil emulsion (e.g., montanide), and proteins (e.g., a cytokine, a complement, GCSF, GM-CSF). MCT and MLT contain point mutations that substantially diminish toxicity without substantially compromising adjuvant activity relative to that of the parent molecules. Immunization of a suitable subject with an immunogenic peptide preparation (e.g., any of the reagents described herein) induces a polyclonal anti-peptide antibody response.

[0414] The term antibody as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules (i.e., molecules that contain an antigen binding site that specifically binds to the peptide (e.g., a peptide described herein)). An antibody that specifically binds to a peptide described herein is an antibody that binds the peptide, but does not substantially bind other molecules in a sample. Examples of immunologically active portions of immunoglobulin molecules include, e.g., F(ab) fragments, F(ab').sub.2 fragments, or any other antibody fragments described herein (see below).

[0415] The anti-peptide antibody can be a monoclonal antibody or a preparation of polyclonal antibodies. The term monoclonal antibody, as used herein, refers to a population of antibody molecules that contain only one species of an antigen binding site capable of immunoreacting with the peptide. A monoclonal antibody composition thus typically displays a single binding affinity for a particular peptide with which it immunoreacts.

[0416] Polyclonal anti-peptide antibodies can be prepared as described above by immunizing a suitable subject with a peptide immunogen. The anti-peptide antibody titer in the immunized subject can be monitored over time by standard techniques, such as with an enzyme linked immunosorbent assay (ELISA) using immobilized peptide. If desired, the antibody molecules directed against the peptide can be isolated from the mammal (e.g., from the blood) and further purified by techniques such as protein A chromatography to obtain the IgG fraction. At an appropriate time after immunization, e.g., when the anti-peptide antibody titers are highest, antibody-producing cells can be obtained from the subject and used to prepare monoclonal antibodies by standard techniques, such as the hybridoma technique originally described by Kohler and Milstein (1975) Nature 256:495-497, the human B cell hybridoma technique (Kozbor et al. (1983) Immunol. Today 4:72), or the EBV-hybridoma technique (Cole et al. (1985), Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96). Any of the many well known protocols used for fusing lymphocytes and immortalized cell lines can be applied for the purpose of generating an anti-peptide monoclonal antibody (see, e.g., Current Protocols in Immunology, supra; Galfre et al. (1977) Nature 266:55052; R. H. Kenneth, in Monoclonal Antibodies: A New Dimension In Biological Analyses, Plenum Publishing Corp., New York, N.Y. (1980); and Lerner (1981) Yale J. Biol. Med., 54:387-402, the disclosures of each of which are incorporated by reference in their entirety).

[0417] As an alternative to preparing monoclonal antibody-secreting hybridomas, a monoclonal anti-peptide antibody can be identified and isolated by screening a recombinant combinatorial immunoglobulin library (e.g., an antibody phage display library) with a peptide described herein to isolate immunoglobulin library members that bind the peptide.

[0418] An anti-peptide antibody (e.g., a monoclonal antibody) can be used to isolate the peptide by techniques such as affinity chromatography or immunoprecipitation. Moreover, an anti-peptide antibody can be used to detect the peptide in screening assays described herein. An antibody can optionally be coupled to a detectable label such as any of those described herein or a first or second member of a binding pair (e.g., streptavidin/biotin or avidin/biotin), the second member of which can be conjugated to a detectable label.

[0419] Non-human antibodies to a target peptide (e.g., a peptide described herein) can also be produced in a non-human host (e.g., a rodent) and then humanized, e.g., as described in U.S. Pat. No. 6,602,503, EP 239 400, U.S. Pat. No. 5,693,761, and U.S. Pat. No. 6,407,213, the disclosures of each of which are incorporated by reference in their entirety.

[0420] Methods for Selecting a Therapy

[0421] Methods for selecting a therapy for a subject with a cancer (e.g., a plasma cell disorder such as multiple myeloma and/or Waldenstrom's macroglobulinemia or any cancer in which XBP1, CD138, or CS1 are expressed (e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, leukemia such as AML or CML)) or with a precancerous condition (e.g., smoldering multiple myeloma) include the steps of: optionally, determining whether one or more cells (e.g., plasma cells) of a subject's cancer express XBP1; and if one or more cells express XBP1, selecting as a therapy for the subject a peptide or composition described herein e.g., a XBP1 peptide or composition comprising a XBP1 peptide described herein.

[0422] Methods for selecting a therapy for a subject with a cancer can include the steps of: optionally, determining whether one or more cells (e.g., plasma cells) of a subject's cancer express CD138; and if one or more cells express CD138, selecting as a therapy for the subject a peptide or composition described herein e.g., a CD138 peptide or composition comprising a CD138 peptide described herein.

[0423] Methods for selecting a therapy for a subject with a cancer can include the steps of: optionally, determining whether one or more cells (e.g., plasma cells) of a subject's cancer express CS-1; and if one or more cells express CS-1, selecting as a therapy for the subject a peptide or composition described herein, e.g., a CS-1 peptide or composition comprising a CS-1 peptide described herein.

[0424] It is understood that where one or more cells (e.g., plasma cells) of a subject's cancer express two or more of XBP1, CD138, and CS-1, a combination of suitable peptides can be delivered to the subject, e.g., via a composition described herein. For example, where one or more cells (e.g., plasma cells) of a subject's cancer are determined to express XBP1 and CD138, the methods for selecting a therapy can include selecting as a therapy for the subject: at least one XBP1 peptide and at least one CD138 peptide described herein, or a composition comprising such peptides.

[0425] Methods for determining whether one or more cells express XBP1, CD138, or CS-1 are known in the art and described above. For example, a biological sample (e.g., a blood sample or lymph node tissue sample) obtained from a subject can be tested using an XBP1-, CD138-, or CS-1-specific antibody made by a method described herein to detect the presence or amount of an XBP1, CD138, or CS-1 polypeptide expressed by a cell (or cell lysate). (See, e.g., the working Examples and Sambrook et al., supra). Methods for assaying a biological sample for the presence or amount of a polypeptide include, e.g., ELISA, immunohistochemistry, flow cytometry, western-blotting, or dot-blotting assays.

[0426] In some embodiments, any of the methods described herein can also include the step of providing a biological sample from a subject and/or obtaining a biological sample from a subject. Suitable biological samples for the methods described herein include any biological fluid, cell, tissue, or fraction thereof, which includes analyte proteins of interest (e.g., XBP1, CD138, or CS-1 proteins). A biological sample can be, for example, a specimen obtained from a subject (e.g., a mammal such as a human) or can be derived from such a subject. For example, a sample can be a tissue section obtained by biopsy, or cells that are placed in or adapted to tissue culture. A biological sample can also be a cell-containing biological fluid such as urine, blood, plasma, serum, saliva, semen, sputum, cerebral spinal fluid, tears, mucus or an aspirate (e.g., a lung or breast nipple aspirate), or such a sample absorbed onto a paper or polymer substrate. A biological sample can be further fractionated, if desired, to a fraction containing particular cell types. For example, a blood sample can be fractionated into serum or into fractions containing particular types of blood cells such as red blood cells or white blood cells (leukocytes). If desired, a sample can be a combination of sample types from a subject such as a combination of a tissue and biological fluid.

[0427] The biological samples can be obtained from a subject, e.g., a subject having, suspected of having, or at risk of developing, a cancer (e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia (e.g., AML or CML), multiple myeloma and/or Waldenstrom's macroglobulinemia). Any suitable methods for obtaining the biological samples can be employed, although exemplary methods include, e.g., phlebotomy, swab (e.g., buccal swab), aspiration, or fine needle aspirate biopsy procedure. Non-limiting examples of tissues susceptible to fine needle aspiration include lymph node, lung, thyroid, breast, and liver. Samples can also be collected, e.g., by microdissection (e.g., laser capture microdissection (LCM) or laser microdissection (LMD)), bladder wash, smear (PAP smear), or ductal lavage.

[0428] After detecting a cancer (e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, multiple myeloma and/or Waldenstrom's macroglobulinemia) or a precancerous condition, e.g., smoldering multiple myeloma, in a subject, e.g., using a method described above, a medical practitioner (e.g., a doctor) can select an appropriate therapeutic modality for the subject (e.g., a therapy comprising one or more of the peptides described herein) by, e.g.: (i) writing a prescription for a medicament; (ii) giving (but not necessarily administering) a medicament to a subject (e.g., handing a sample of a prescription medication to a patient while the patient is at the physician's office); (iii) communication (verbal, written (other than a prescription), or electronic (email, an electronic post to a secure site)) to the patient of the suggested or recommended therapeutic modality (e.g., a therapy comprising one or more of the peptides described herein); or (iv) identifying a suitable therapeutic modality for a subject and disseminating the information to other medical personnel, e.g., by way of patient record. The latter (iv) can be useful in a case where, e.g., more than one therapy or therapeutic agent are to be administered to a patient by different medical practitioners.

[0429] After detecting the presence or amount of XBP1, CD138, or CS-1 in a subject (using any of the methods above); and/or selecting a therapy for the subject, a medical practitioner (e.g., a doctor) can administer the appropriate therapeutic modality to the subject. Methods for administering a therapy comprising one or more of the peptides described herein as detailed above.

[0430] In addition, a medical practitioner can also select, prescribe and/or administer one or more additional therapeutic agents to treat a cancer or one or more medicaments to treat side-effects of an anti-cancer agent. Suitable chemotherapeutic agents for treating multiple myeloma and/or Waldenstrom's macroglobulinemia include, e.g., melphalan, cyclophosphamide, vincristine, doxorubicin, prednisone, dexamethasone, proteosome inhibitors (e.g., bortezomib), thalidomide, or lenalidomide.

[0431] Side effects of anti-cancer agents include, e.g., anemia, gastrointestinal symptoms (e.g., nausea, vomiting, diarrhea), leukopenia (decreased number of white blood cells, which may cause infection), temporary hair loss, or thrombocytopenia (decreased number of platelets, which may cause bleeding). Thus, a doctor can prescribe or administer to a subject a chemotherapeutic agent such as vincristine along with an anti-anemia medicament such as epoetin alpha (e.g., Procrit.RTM. or Epogen.RTM.).

[0432] Kits and Articles of Manufacture

[0433] The disclosure also features a variety of kits. The kits can include, e.g., one or more (e.g., one, two, three, four, five, six, seven, eight, nine, or 10 or more) of any of the peptides or compositions (or expression vectors containing nucleic acid sequences encoding one or more peptides) described herein; and instructions for administering the peptide or composition to a subject. The kit can include one or more pharmaceutically acceptable carriers and/or one or more immune stimulating agents and/or one or more immune modulating agents. The immune stimulating agents can be, e.g., a T helper epitope, an altered peptide ligand, or an adjuvant. In one embodiment, the immune stimulating agent can be a combination of carboxymethylcellulose, polyinosinic-polycytidylic acid, and poly-L-lysine double-stranded RNA (e.g., poly IC-LC, e.g., hiltonol); a water-and-oil emulsion (e.g., montanide); or a protein (e.g., a cytokine, a complement, GCSF, GM-CSF). In one embodiment, the immune modulating agent is a protein, e.g., an antibody which activates the immune system (e.g., an anti-CTLA4 antibody, e.g., ipilimumab or tremelimumab, an anti-PD-1 antibody, an anti-PDL-1 antibody); a small molecule adjuvant (e.g., thalidomide or a thalidomide derivative, e.g., lenalidomide).

[0434] The kits can also contain one or more therapeutic agents, diagnostic agents, or prophylactic agents. The one or more therapeutic, diagnostic, or prophylactic agents include, but are not limited to: (i) an agent that modulates inflammatory responses (e.g., aspirin, indomethacin, ibuprofen, naproxen, steroids, cromolyn sodium, or theophylline); (ii) an agent that affects renal and/or cardiovascular function (e.g., furosemide, thiazide, amiloride, spironolactone, captopril, enalapril, lisinopril, diltiazem, nifedipine, verapamil, digoxin, isordil, dobutamine, lidocaine, quinidine, adenosine, digitalis, mevastatin, lovastatin, simvastatin, or mevalonate); (iii) drugs that affect gastrointestinal function (e.g., omeprazole or sucralfate); (iv) antibiotics (e.g., tetracycline, clindamycin, amphotericin B, quinine, methicillin, vancomycin, penicillin G, amoxicillin, gentamicin, erythromycin, ciprofloxacin, doxycycline, streptomycin, gentamicin, tobramycin, chloramphenicol, isoniazid, fluconazole, or amantadine); (v) anti-cancer agents (e.g., cyclophosphamide, methotrexate, fluorouracil, cytarabine, mercaptopurine, vinblastine, vincristine, doxorubicin, bleomycin, mitomycin C, hydroxyurea, prednisone, tamoxifen, cisplatin, or decarbazine); (vi) immunomodulatory agents (e.g., interleukins, interferons (e.g., interferon gamma (IFN-.gamma.), granulocyte macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor alpha (TNF.alpha.), tumor necrosis factor beta (TNF.beta.), cyclosporine, FK506, azathioprine, steroids); (ix) drugs acting on the blood and/or the blood-forming organs (e.g., interleukins, G-CSF, GM-CSF, erythropoietin, heparin, warfarin, or coumarin); or (vii) hormones (e.g., growth hormone (GH), prolactin, luteinizing hormone, TSH, ACTH, insulin, FSH, CG, somatostatin, estrogens, androgens, progesterone, gonadotropin-releasing hormone (GnRH), thyroxine, triiodothyronine); hormone antagonists; agents affecting calcification and bone turnover (e.g., calcium, phosphate, parathyroid hormone (PTH), vitamin D, bisphosphonates, calcitonin, fluoride).

[0435] Also featured are articles of manufacture that include: a container; and a composition contained within the container, wherein the composition comprises an active ingredient for inducing an immune response in a mammal (e.g., a human), wherein the active ingredient comprises one or more (e.g., two, three, four, five, six, seven, eight, nine, or 10 or more) of any of the peptides described herein, and wherein the container has a label indicating that the composition is for use in inducing an immune response in a mammal (e.g., any of the mammals described herein). The label can further indicate that the composition is to be administered to a mammal having, suspected of having, or at risk of developing, a cancer, e.g., lung cancer, liver cancer, bile duct cancer, stomach cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, multiple myeloma, smoldering multiple myeloma and/or Waldenstrom's macroglobulinemia. The composition of the article of manufacture can be dried or lyophilized and can include, e.g., one or more solutions (and/or instructions) for solubilizing a dried or lyophilized composition.

[0436] The articles of manufacture can also include instructions for administering the composition to the mammal (e.g., as described above).

[0437] The following examples are intended to illustrate, not limit, the invention.

Examples

Example 1: Materials and Methods

[0438] Cell Lines.

[0439] The multiple myeloma cell lines: McCAR, MM1S and U266, were obtained from the American Type Culture Collection (ATCC; Manassas, Va.). The human acute myeloid leukemia (AML) cell line, ML-2, was kindly provided by Dr. Y. Matsuo, Fujisaki Cell Center, Okayama, Japan. The T2 cell line, which is a human B and T cell hybrid expressing HLA-A2.1 molecules (Zweerink et al., (1993) J Immunol. 150(5):1763-71), was provided by Dr. J. Molldrem (University of Texas M. D. Anderson Cancer Center, Houston, Tex.) and was used as a source of antigen presenting cells (APCs). K562-A*0201 cells were provided by Karen Anderson (Dana Farber Cancer Institute, Boston, Mass.) and used in immune monitoring assays to present individual peptides to the CTL. Various cancer cells including LnCap, VCap, MB231, MCF7, BT474, LS180, SW480, WiDRr, OCI, U937, HEL, UT7, HL60, Nomo1, and THP1 were obtained from the ATCC. All cell lines were cultured in RPMI-1640 medium (Gibco-Life Technologies, Rockville, Md.) supplemented with 10% fetal calf serum (FCS; BioWhittaker, Walkersville, Md.), 100 IU/ml penicillin and 100 .mu.g/ml streptomycin (Gibco-Life Technologies).

[0440] Reagents.

[0441] Mouse anti-human CD80 or CD83 monoclonal antibodies (mAb) conjugated with phycoerythrin (PE) were purchased from Immunotech (Hialeigha, Fla.). Mouse anti-human CD3, CD4, CD8, CCR7, CD45RO, CD69, CD107.alpha., IFN-.gamma., and HLA-A2 mAbs conjugated with FITC, PE, PerCP, PerCP-Cy5.5, APC, Pacific Blue, APC-H7 or PE-Cy7 were purchased from Becton Dickinson (BD)/Pharmingen or BD/Biosciences (San Diego, Calif.). Recombinant human IL-2, IL-4, IFN-.alpha. and TNF-.alpha. were purchased from R&D Systems (Minneapolis, Minn.), and GM-CSF was obtained from Immunex (Seattle, Wash.).

[0442] Synthetic Peptides.

[0443] Influenza virus protein matrix peptide.sub.58-66 (GILGFVFTL; SEQ ID NO:25) and MAGE-3 peptide (FLWGPRALV; SEQ ID NO:26) were used as control HLA-A2-binding peptides. Six native non-spliced XBP1 peptides: XBP1.sub.118-126 (LLREKTHGL; SEQ ID NO:1); XBP1.sub.185-193 (NISPWILAV (SEQ ID NO:2)); XBP1.sub.190-198 (ILAVLTLQI (SEQ ID NO:3)); XBP1.sub.193-201 (VLTLQIQSL (SEQ ID NO:4)); XBP1.sub.111-119 (KLLLENQLL (SEQ ID NO:5)); XBP1.sub.94-102 (RMSELEQQV (SEQ ID NO:27)); three native spliced XBP1 peptides including SP XBP1.sub.197-205 (GILDNLDPV (SEQ ID NO:7)); SP XBP1.sub.194-202 (ILLGILDNL (SEQ ID NO:8)); SP XBP1.sub.368-376 (ELFPQLISV (SEQ ID NO:9)); heteroclitic XBP1 (YISPWILAV (SEQ ID NO:6)); and heteroclitic spliced XBP1 (YILDNLDPV (SEQ ID NO: 24)); and YLFPQLISV (SEQ ID NO:10)) peptides were designed and examined as potential HLA-A2- binding peptides. As used herein, "heteroclitic" (e.g., a heteroclitic peptide) refers to a form of a peptide in which one or more amino acids have been modified from a wild-type or original sequence in order to produce a peptide that is more immunogenic than the corresponding wild-type peptide. For example, in the exemplary heteroclitic peptides described immediately above, the bolded amino acids indicate the amino acids that are modified from the wild-type sequence of XBP1.

[0444] Four native CD138 peptides: CD138.sub.256-264 (VIAGGLVGL (SEQ ID NO:11)); CD138.sub.260-268 (GLVGLIFAV (SEQ ID NO:12)); CD138.sub.5-13 (ALWLWLCAL (SEQ ID NO:13)); and CD138.sub.7-15 (WLWLCALAL (SEQ ID NO:14)) were designed and examined as potential HLA-A2-binding peptides.

[0445] Four native CS1 peptides: CS1-P1: CS.sub.1236-245 (LLLSLFVLGL (SEQ ID NO:15)); CS1-P2: C51.sub.236-247 (SLFVLGLFL (SEQ ID NO:16)); CS1-P3: C51.sub.232-240 (LLVPLLLSL (SEQ ID NO:17)); and CS1-P4: C51.sub.6-17 (TLIYILWQL (SEQ ID NO:18)) were designed (using three different databases RANKPEP, BIMAS and NetMHC) and examined as potential HLA-A2-binding peptides. (See, e.g., Reche et al. (2002) Human Immunology 63:710-709).

[0446] XBP-1 and CD138 peptides were synthesized (Biosynthesis, Lewisville, Tex.) by standard FMOC (9-fluorenylmethyl-oxycarbonyl) chemistry, purified to >85% using reverse-phase chromatography and validated by mass-spectrometry for molecular weight. CS1 peptides were synthesized by New England Peptides LLC with greater than 95% purity.

[0447] Heteroclitic XBP1 US.sub.185-193 (YISPWILAV) (SEQ ID NO: 6), heteroclitic XBP1 SP.sub.368-376 (YLFPQLISV) (SEQ ID NO: 10), native CD138.sub.260-268 (GLVGLIFAV) (SEQ ID NO: 12), and native CS.sub.1239-247 (SLFVLGLFL) (SEQ ID NO: 16) peptides were derived from XBP1 unspliced (US), XBP1 spliced (SP), CD138 and CS1 antigens, respectively. Influenza virus matrix protein.sub.58-66 (GILGFVFTL) (SEQ ID NO: 25) and CMV pp65 (NLVPMVATV) (SEQ ID NO: 28) were selected as HLA-A2-specific control peptides. All peptides were synthesized by standard fmoc (9-fluorenylmethyl-oxycarbonyl) chemistry, purified to >90% using reverse-phase chromatography, and validated by mass-spectrometry for molecular weight (Biosynthesis, Lewisville, Tex.). Lyophilized peptides were dissolved in DMSO (Sigma, St. Louis, Mo.), diluted in AIM-V medium (Gibco-Life Technologies), and stored at -140.degree. C.

[0448] Peptide Binding Assay.

[0449] A cocktail of four HLA-A2 peptides, heteroclitic XBP1 US.sub.185-163, heteroclitic XBP1 SP.sub.368-376, CD138.sub.260-268 and CS1.sub.239-247, was evaluated for binding affinity using the T2 cell line. In the assay, T2 cells were washed three times, resuspended in serum-free AIM-V medium (Gibco-Life Technologies) to a final concentration of 1.times.10.sup.6 cells/ml, and transferred into a 48-well tissue culture plate. The cells were pulsed with a cocktail of the four peptides at total peptide concentrations ranging from 0-50 .mu.g/ml plus 3 .mu.g/ml human 02-microglobulin (Sigma, St Louis, Mo.), and incubated at 37.degree. C., 5% CO.sub.2 in humidified air. Following overnight incubation, cells were washed, stained with mouse anti-human HLA-A2-FITC mAb for 15 minutes at 4.degree. C., and analyzed using a FACSCanto.TM. II flow cytometer (Becton Dickinson, San Jose, Calif.).

[0450] Peptide Stability Assay.

[0451] The multipeptide cocktail was examined for HLA-A2 stability over time. After overnight incubation of T2 cells pulsed with the multipeptide cocktail (25 .mu.g/ml; 6.25 .mu.g/ml/peptide), the cells were washed to remove unbound peptide and incubated with 10 .mu.g/ml Brefeldin A (Sigma) at 37.degree. C. and 5% CO.sub.2 for 1 hour to block cell surface expression of newly synthesized HLA-A2 molecules. Peptide/HLA-A2 complex stability was measured at 0, 2, 4, 6 and 14 hours post-BFA treatment by staining cells with mouse anti-human HLA-A2-FITC mAb and analyzing by flow cytometry.

[0452] Generation of Monocyte-Derived Mature Dendritic Cells.

[0453] Peripheral blood mononuclear cells (PBMC) were isolated by standard density gradient centrifugation over Ficoll-Paque.TM. Plus (Amersham Pharmacia Biotech AB, Uppsala Sweden) from leukopaks obtained from HLA-A2.sup.+ normal individuals. To generate dendritic cells (DC), monocytes isolated as the adherent cell fraction were cultured for 7 days in the presence of 1,000 U/ml GM-CSF and 1,000 U/ml IL-4 in RPMI-1640 medium (Gibco-Life Technologies) supplemented with 10% FCS. Fresh media plus GM-CSF and IL-4 was added to the cultures every other day. Mature DC (mDC) were obtained by adding 1,000 U/ml IFN-.alpha. plus 10 ng/ml TNF-.alpha. along with fresh GM-CSF and IL-4 in 10% FCS-RPMI on day 7 and incubating for an additional three days.

[0454] Isolation of CD3.sup.+ T Cells.

[0455] CD3.sup.+ T cells were obtained by negative selection from the non-adherent cell fraction using the EasySep.RTM. magnet and Robosep.RTM. from StemCell Technologies (Vancouver, Canada). In brief, T cell enrichment was accomplished by depletion of non-CD3 T cells including B cells, monocytes, NK cells, erythroid cells, platelets and basophils by labeling in bispecific tetrameric antibody complexes which are directed against CD14, CD16, CD19, CD20, CD36, CD56, CD66b, CD123, and glycophorin A. After the removal of magnetically labeled unwanted cells, the enriched CD3.sup.+ T cells were washed and examined by flow cytometry.

[0456] Isolation of Primary CD138.sup.+ Cells from Bone Marrow Mononuclear Cells of MM Patients.

[0457] Bone marrow mononuclear cells (BMMC) were isolated by standard density gradient centrifugation over Ficoll-Paque.TM. Plus from bone marrow cells obtained from MM patients. CD138.sup.+ MM cells were isolated from BMMC using RoboSep.RTM. CD138 positive immunomagnetic selection technology (StemCell Technologies).

[0458] Induction of Peptide-Specific CTL.

[0459] CTL specific for individual peptides (peptide-specific CTL) or multiple peptides (MP-CTL) were generated ex vivo by repeated stimulation of CD3.sup.+ T lymphocytes obtained from normal HLA-A2.sup.+ or HLA-A24+ donors (see FIG. 25). In brief, APC (mDC or T2 cells) were pulsed overnight at 37.degree. C. and 5% CO.sub.2 in humidified air with either individual peptides or a cocktail of heteroclitic XBP1 US.sub.185-193, heteroclitic XBP1 SP.sub.368-376, CD138.sub.260-268 and CS1.sub.239-247 peptides (25 .mu.g/ml total peptide). The loaded APC were harvested, washed, irradiated at 20 Gy, and resuspended in AIM-V medium supplemented with 10% human AB serum (Gemni Bio Products, West Sacramento, Calif.). Irradiated mp-pulsed mDC were used to prime autologous CD3.sup.+ T cells at a 1:20 APC/mp-to-CD3.sup.+ T cell ratio in AIM-V medium supplemented with 10% human AB serum. The cultures were restimulated every seven days with irradiated APC/mp for a total of 4 cycles to generate CTL specific to the mp. IL-2 (50 U/ml) was added to the cultures two days after the second stimulation and replenished until the culture is completed.

[0460] Phenotypic Analysis of the XBP1-CTL, CD138-CTL, or Target Cells.

[0461] One week after the fourth stimulation, MP-CTL and control T cells were evaluated for total CD3.sup.+CD8.sup.+ T cells or naive, effector memory, and activated CD3.sup.+CD8.sup.+ T cells by staining them with CD3-PacBlue, CD8-APC-H7, CCR7-PeCy7, CD45RO-PE and/or CD69-PerCP mAbs at 4.degree. C. for 30 minutes. After staining, the cells were washed, fixed in 2% paraformaldehyde-PBS, and analyzed by flow cytometry.

[0462] Western Blotting.

[0463] Approximately 100 .mu.g protein lysate from each cell line (U266, McCAR, ML-2, and MM1S) was suspended in Laemmli's sample buffer (0.1 M Tris-HCl buffer, pH 6.8, containing 1% sodium dodecyl sulfate (SDS), 0.05% .beta.-mercaptoethanol, 10% glycerol and 0.001% bromophenol blue), boiled for two minutes and subjected to 8-16% gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for two hours (Xcell Surelock Mini Cell, Invitrogen, Carlsbad, Calif.) at 80 V. A protein ladder (a mixture of proteins of known molecular weight) was used as a size marker in the gel to determine the molecular weight of the peptides (Invitrogen, Carlsbad, Calif.). Gels were electroblotted onto nitrocellulose membranes (Trans-Blot, 0.2 micron transfer membrane, Bio-Rad Laboratories, CA) at 40 V for two hours in a Tris-glycine buffer. Transfer of the proteins onto the nitrocellulose membrane was confirmed by Ponceau S staining. Incubation of the membrane with mouse anti-human XBP1 antibody or an anti-human CD138 antibody was performed for one hour in phosphate buffered saline and Tween 20 (PBST) containing 1% BSA with constant rocking. The membrane was washed three times with PBST and incubated in anti-mouse IgG-horseradish peroxidase conjugates for one hour in PBST containing 3% non-fat dry milk. After washing, specific proteins were detected using enhanced chemiluminescence, according to the instructions provided in the product manual (Amersham Life Sciences Inc., Arlington Heights, Ill.).

[0464] IFN-.gamma. ELISA.

[0465] IFN-.gamma. release by XBP1-CTL, CD138-CTL, or CS1-CTL following co-culture with multiple myeloma (MM) cells (McCAR, MM1S), acute myelogenous leukemia (AML) cells (ML-2), or T2 cells (above) was measured using a human IFN-.gamma. ELISA kit from BD Biosciences (San Diego, Calif.) (see FIG. 27). Briefly, dilutions of purified IFN-.gamma. as standards or CTL supernates were transferred into wells of a 96-well plate pre-coated with a monoclonal anti-human IFN-.gamma. capture antibody and incubated for 2 hours at room temperature. After several washes, a buffer containing a detection antibody and avidin-horseradish peroxidase conjugate was added to each well and incubated for one hour at room temperature. The wells were washed and then to each well was added a horseradish peroxidase substrate solution and incubated at room temperature for 30 minutes. Stop solution was added to each well and the absorbance was determined at 450 nm was determined with a PerkinElmer Wallac Victor2 counter (PerkinElmer, Wellesley, Mass.). The amount of cytokine present in the CTL culture supernatant was calculated based on the IFN-.gamma. standard curve.

[0466] Cell Proliferation by Carboxy Fluorescein Succinimidyl Ester (CFSE) Tracking.

[0467] CTL proliferation was measured after co-culture with multiple myeloma (MM) cells (McCAR, MM1S), acute myelogenous leukemia (AML) cells (ML-2), or T2 cells (above) (see FIGS. 27 and 32). Individual XBP1-CTL, CD138-CTL, CS1-CTL or multi-peptide generated CTL were washed twice in PBS (Gibco-BRL) and resuspended in RPMI-1640 culture medium at a concentration of 1.times.10.sup.6 cells/ml. CFSE (Molecular Probes, Eugene, Oreg.) in the form of a 5 mM stock solution in DMSO was added to the CTL to give a final concentration of 5 .mu.M and incubated for 10 minutes at 37.degree. C. in a CO.sub.2 incubator protected from light. After incubation, a volume of ice-cold PBS (containing 2% FCS) equivalent to five-times the volume of the CTL cells was added to the cells to quench the reaction. Cells were incubated for 5 minutes on ice, centrifuged and resuspended in fresh PBS (containing 2% FCS) after a total of three washings. CFSE-labeled T cells were adjusted to a concentration of 2.times.10.sup.6 cells/ml with RPMI culture medium, stimulated with 2.times.10.sup.5 cells/ml primary multiple myeloma cells, MM cell lines, various cancer cell lines or K562-A2 cells presenting individual peptides. The stimulated CFSE-labeled cells were examined by flow cytometry.

[0468] Cytotoxicity Assay.

[0469] The cytotoxic activities of individual XBP1-CTL, CD138-CTL, CS1-CTL or multi-peptide generated CTL were measured by a calcein release assay as described by, e.g., Roden et al. (1999) J. Immunol Methods 226:29-41. Briefly, target cells (3.times.10.sup.5 cells) including T2, U266 cells, McCAR cells, ML-2 cells, MM1S cell lines or primary multiple myeloma cells were incubated in serum-free culture medium containing 10 mM calcein-AM (Molecular Probes) for 30 minutes at 37.degree. C., washed three times in cold PBS with 5% FCS, and incubated with effector cells (5.times.10.sup.3 cells/well) at various effector:target cell ratios in 96-well U-bottom microtiter plates (triplicate wells/sample). Plates were incubated for 3 hours at 37.degree. C. and 5% CO.sub.2. After incubation, the cells were pelleted by centrifugation at 1,000 rpm for 5 minutes and 100 .mu.l of the supernatant was transferred to the wells of 96-well flat-bottomed microtiter plates (Nunc) and the calcein-release was measured as the amount of fluorescence released from the cells (using a VICTOR.sup.2-1420 multilabel counter (PerkinElmer, Boston, Mass.)). Maximum calcein release was obtained from detergent-released target cell counts and spontaneous release from target cell counts in the absence of effector cells. Cellular cytotoxicity was calculated as follows: % Specific Lysis=[(experimental release-spontaneous release) (maximum release-spontaneous release)].times.100.

[0470] CD107.alpha. Degranulation Assay.

[0471] CD107.alpha. degranulation was measured after co-culture with multiple myeloma (MM) cells (McCAR, MM1S), acute myelogenous leukemia (AML) cells (ML-2), or T2 cells (above) (see FIGS. 27 and 30). The CD107.alpha. degranulation assay, a measure of cytotoxic activity, was performed as described by Betts et al. (2003) and Mittendorf et al. (2005) with minor modifications as detailed below. Primary multiple myeloma cells, MM cell lines, various cancer cell lines or K562-A2 cells pulsed with individual peptide were co-cultured at various effector:target ratios with CTL. A 10 .mu.l aliquot of each of CD107.alpha. and CD107b (both conjugated to the detectable label FITC) was added to each well at the same time of adding the CD138-CTL. The plate containing the cells was centrifuged for 5 minutes at 1000 rpm and incubated at 37.degree. C. for one hour. Following incubation, Brefeldin A and Monensin was added to each well and the cells were incubated at 37.degree. C. for an additional four hours. The cells were collected and washed and stained with fluorochrome conjugated anti-human MABs The cells were washed and analyzed by flow cytometry.

[0472] CD107.alpha. Upregulation and Intracellular IFN-.gamma. Production.

[0473] CD107.alpha. upregulation and IFN-.gamma. production were measured after co-culture with multiple myeloma (MM) cells (McCAR, MM1S), acute myelogenous leukemia (AML) cells (ML-2), or T2 cells (above) (see FIGS. 27 and 30). CD107.alpha. degranulation and IFN-.gamma. producing CD8.sup.+ CTL were determined by cell surface marker and intracellular cytokine staining by flow cytometry. Briefly, 1.times.10.sup.6 responder cells (MP-CTL, Control T cells) were stimulated with 1.times.10.sup.6 stimulator cells (HLA-A2.sup.+ McCAR or U266 MM cell lines or K562-A*0201 cells pulsed with respective peptide). CD107.alpha. mAb was added to the cultures, and cells were placed into a 37.degree. C., 5% CO.sub.2 incubator. After 1 hour incubation, CD28/CD49d mAb (BD) and protein transport inhibitors Brefeldin A (BD) and Monensin (BD) were added to the cell cultures and incubated for an additional 5 hours. As a baseline control, MP-CTL were cultured in media with CD28/CD49d mAb, Brefeldin A and Monensin alone, without further stimulation. After incubation, the cells were harvested, washed, and surface stained with CD3-PacBlue and CD8-APC-H7, CCR7-PeCy7, CD45RO-PE and/or CD69-PerCP anti-human mAbs for 30 minutes. Cells were then permeabilized, fixed using Cytofix/Cytoperm (BD), and stained with anti-IFN-.gamma. FITC mAb for 45 minutes to detect intracellular cytokine production. Lastly, cells were washed with Perm/Wash solution (BD), fixed in 2% paraformaldehyde, and acquired by flow cytometry.

[0474] IL-2 Production Assay.

[0475] CD107.alpha. degranulation and IL-2 producing CD3+CD8+ CTL were identified by cell surface marker and intracellular cytokine staining by flow cytometry. Briefly, the peptide-specific CTL or control T cells were stimulated with each specific stimulator in the presence of CD107.alpha. mAb. After 1 hour incubation, CD28/CD49d mAb (BD), as well as protein transport inhibitors Brefeldin A and Monensin, were added to the cultures and incubated for an additional 5 hours. As a baseline control, the CTL were cultured in media with CD28/CD49d mAb, Brefeldin A, and Monensin alone. After incubation, cells were stained with CD3-PacBlue and CD8-APC-H7 anti-human mAbs, followed by fixation/permeabilization and stained with anti-IL-2 APC anti-human mAb to detect intracellular cytokine production. After the staining, cells were washed with Perm/Wash solution three times, fixed in 2% paraformaldehyde, and analyzed by flow cytometry.

[0476] Statistical Analysis.

[0477] Results are presented as mean.+-.SE. Groups were compared using unpaired Student's t-test. Differences were considered significant when p<0.05.

Example 2: A Multipeptide (MP) Cocktail of XBP1 Unspliced, XBP1 Spliced, CD138 and CS1-Specific Peptides Displays High HLA-A2 Binding Affinity and Stability

[0478] The four immunogenic peptides, heteroclitic XBP1 US.sub.185-193 (YISPWILAV, SEQ ID NO: 6), heteroclitic XBP1 SP.sub.368-376 (YLFPQLISV, SEQ ID NO: 10), native CD138.sub.260-268 (GLVGLIFAV, SEQ ID NO: 12), and native CS1.sub.239-247 (SLFVLGLFL, SEQ ID NO: 16) (Table 1), have been individually demonstrated to induce immune response. Here we have evaluated them as a MP cocktail. The HLA-A2-specific binding and stability of the MP cocktail was evaluated by measuring the upregulation of HLA-A2 molecules on T2 cells by flow cytometry (27). The peptide binding assay demonstrated an increase in the HLA-A2 mean fluorescence intensity (MFI) on T2 cells in a dose-dependent manner (0-50 .mu.g/ml), reaching a plateau at a total peptide concentration of 25 .mu.g/ml (6.25 .mu.g/peptide/ml; MFI: 10,787.33.+-.2,371.71) similar to the highest total peptide concentration, 50 .mu.g/ml (MFI: 10,889.33.+-.2,888.48) (FIG. 1a). Therefore, a MP concentration of 25 .mu.g/ml (6.25 .mu.g/peptide/ml) was selected for evaluation of HLA-A2 binding stability.

[0479] In the peptide binding stability assay, T2 cells were pulsed overnight with 25 .mu.g/ml of the MP cocktail, washed to remove unbound peptides, and then treated with Brefeldin A (BFA) to block cell surface expression of newly synthesized HLA-A2 molecules. T2 cells were then evaluated for their HLA-A2 MFI at 0, 2, 4, 6 or 14 hrs post-BFA treatments. Flow cytometry analysis demonstrated that the stability of MP cocktail was highly maintained up to 6 hrs post-BFA treatment (MFI: 0 hr=9,726.00.+-.1,373.24, 2 hr=9,132.33.+-.1,435.51, 4 hr: 9,125.33.+-.1,130.62, 6 hr: 8,818.67.+-.413.50) (FIG. 1b). At 14 hr post-BFA treatment, the HLA-A2-specific affinity of MP cocktail was lower, but was greater (MFI: 6,793.67.+-.1,617.01) than the affinity of the control influenza virus matrix protein (IVMP).sub.58-66 peptide (MFI: 4,921.33.+-.1,428.16). Based on these results, we confirmed a high level of HLA-A2-specific affinity and stability of the MP cocktail and proceeded to further evaluate the cocktail for its immunogenicity and ability to induce MM-specific CTL.

TABLE-US-00005 TABLE 4 Native and heteroclitic epitopes evaluated as a multipeptide for targeting MM TAA Identification Type Sequence XBP1- XBP1.sub.185-193 Hetero- YISPWILAV unspliced clitic (SEQ ID NO: 6) XBP1- XBP1-SP.sub.368-376 Hetero- YLFPQLISV spliced clitic (SEQ ID NO: 10) CD138 CD138.sub.260-268 Native GLVGLIFAV (SEQ ID NO: 12) CS1 CS1.sub.239-247 Native SLFVLGLFL (SEQ ID NO: 16)

Example 3: Multipeptide-Specific CTL Display a Distinct Phenotype Representing Specific T Cell Subtypes

[0480] The MP-CTL were generated by stimulating HLA-A2.sup.+ normal donor's T cells weekly with APC pulsed with the MP cocktail (25 .mu.g/ml total; 6.25 .mu.g/ml/peptide). One week after the fourth stimulation, the resulting MP-CTL were evaluated for their phenotype and functional activity. Flow cytometry analyses showed that the MP-CTL contained a higher proportion of CD3.sup.+CD8.sup.+ T cells (donor 1: 86%, donor 2: 74%) compared to control T cell cultures (donor 1: 25%, donor 2: 25%; FIG. 2). We also observed distinct phenotypic changes in the CD3.sup.+CD8.sup.+ T cell subset within the MP-CTL. The frequency of effector memory T cells (EM: CD45RO.sup.+CCR7.sup.-/CD3.sup.+CD8.sup.+) was increased (Donor 1: Control 5% vs. MP-CTL 44%, Donor 2: Control 4% vs. MP-CTL 35%), associated with a corresponding decrease in naive T cells (CD45RO.sup.-CCR7.sup.+/CD3.sup.+CD8.sup.+) (Donor 1: Control 74% vs. MP-CTL 8%, Donor 2: Control 60% vs. MP-CTL 6%). In addition, we observed an increase in the frequency of activated CD69.sup.+/CD3.sup.+CD8.sup.+ T cells within the MP-CTL as compared to the control T cell cultures (Donor 1: Control 3% vs. MP-CTL 39%, Donor 2: Control 5% vs. MP-CTL 13%; FIG. 2). Thus, these results demonstrate that repeated stimulation of CD3.sup.+ T cells with the MP cocktail specific to XBP1, CD138 or CS1 results in distinct phenotypic changes and expansion of the CD3.sup.+/CD8.sup.+ T cell subsets characteristic of antigen-specific CTL.

Example 4: Multipeptide-Specific CTL Include a High Proportion of CD8.sup.+ CTL Producing IFN-.gamma. in Response to HLA-A2.sup.+ MM Cells

[0481] MP-CTL were analyzed for their ability to produce intracellular IFN-.gamma. upon stimulation with the HLA-A2.sup.+ MM cell lines by flow cytometry. Both EM (CD45RO.sup.+CCR7.sup.-) and activated (CD69.sup.+) CD3.sup.+CD8.sup.+ T cells within the MP-CTL produced IFN-7 in response to HLA-A2.sup.+ MM cell lines (FIG. 3). The frequency of IFN-.gamma. producing cells was increased upon stimulation with either McCAR cells [Donor 1: Control vs. MP-CTL--0% vs. 4.7% EM cells, 0.8% vs. 6.1% activated cells; Donor 2: 0.2% vs. 2.7% EM cells, 1% vs. 3.9% activated cells] or U266 cells [Donor 1: Control vs. MP-CTL--0% vs. 8% EM cells, 0% vs. 11.2% activated cells; Donor 2: 0.4% vs. 2.9% EM cells, 1.3% vs. 3.0% activated cells]. The naive (CD45RO.sup.-CCR7.sup.+) CD3.sup.+CD8.sup.+ T cells within the MP-CTL showed a minimum level of IFN-.gamma.-production when stimulated with the MM cell lines (data not shown).

Example 5: Multipeptide-Specific CTL Show Cell Proliferation in Response to HLA-A2.sup.+ MM Cells

[0482] The function of the MP-CTL was analyzed using a CFSE-proliferation assay. MP-CTL proliferation was measured on day 5 as a decrease in fluorescence of the CFSE-labeled MP-CTL (Q1-gated cells) following stimulation with HLA-A2.sup.+ MM primary cells or cell lines (FIG. 4). The MP-CTL showed a high level of cell proliferation in response to CD138.sup.+ primary cells obtained from three different HLA-A2.sup.+ MM patients (proliferating cells: 33%, 29% or 41%). In addition, the MP-CTL demonstrated a high level of cell proliferation in response to HLA-A2.sup.+ MM cell lines including McCAR (proliferating cells: 57%) and U266 (proliferating cells: 49%). MP-CTL cultured in media alone displayed a low level (5%) of proliferation. Taken together, these data demonstrate the functional activity of MP-CTL through their ability to proliferate when stimulated with either HLA-A2.sup.+ primary MM cells or MM cell lines.

Example 6: Multipeptide-Specific CTL Induce Specific Lysis of HLA-A2.sup.+ MM Cells

[0483] We evaluated the cytotoxic activity of MP-CTL using a 4-hour calcein-release assay. The MP-CTL generated from different HLA-A2.sup.+ donors' CD3.sup.+ T cells were evaluated for their cytotoxic activity against HLA-A2.sup.+ MM primary cells or cell lines (FIG. 5). The HLA-A2.sup.+ primary MM cells were effectively lysed by the MP-CTL ([Donor A MP-CTL; Patient #1: 6-29%, Patient #2: 0-49%], [Donor B MP-CTL; Patient #1: 0-17%, Patient #2: 0-15%). In addition, MP-CTL demonstrated a high level of cytotoxic activity against the U266 cells (Donor A MP-CTL: 0-85%, Donor B MP-CTL: 2-44%) and McCAR cells (Donor A MP-CTL: 0-13%, Donor B MP-CTL: 0-79%:) at the various Effector:Target cell ratios. Compared to the MP-CTL, control CD3.sup.+ T cells from the same donors showed a significantly lower level of cytotoxicity against HLA-A2.sup.+ MM primary cells or cell lines. In addition, the MP-CTL did not lyse antigen mismatched or MHC mismatched tumor cells including a HLA-A2.sup.+ breast cancer cell line (MCF-7), HLA-A2.sup.- MM cell line (MM1S) or HLA-A2.sup.- primary cells from three different MM patients (data not shown). Taken together, these data confirm the HLA-A2-restricted and antigen-specific cytotoxic activity of MP-CTL.

Example 7: Multipeptide-Specific CTL Generate Individual Immune Responses to Each Relevant Peptide

[0484] MP-CTL were analyzed for their ability to degranulate (CD107.alpha. expression) and produce intracellular IFN-.gamma. in response to each relevant peptide including heteroclitic XBP1 US.sub.185-193 (YISPWILAV) (SEQ ID NO: 6), heteroclitic XBP1 SP.sub.368-376 (YLFPQLISV) (SEQ ID NO: 10), native CD138.sub.260-268(GLVGLIFAV) (SEQ ID NO: 12), and native CS1.sub.239-247 (SLFVLGLFL) (SEQ ID NO: 16). The analyses were performed by measuring the specific MP-CTL responses to K562-A*0201 cells pulsed with the respective peptide. As controls, we used no peptide-pulsed K562-A*0201 cells or K562-A*0201 cells pulsed with an irrelevant HLA-A2-specific CMV pp65 (NLVPMVATV) (SEQ ID NO: 28) peptide. FIG. 6a shows a representative flow cytometric analysis of the peptide-specific response from Donor A MP-CTL. The MP-CTL showed a high proportion of CD107.alpha..sup.+IFN.gamma..sup.+/CD3.sup.+CD8.sup.+ T cells (gated Q2) in response to XBP1 US (2.7%), CD138 (1.7%), and CS1 (12.5%) peptides, but not to XBP1 SP (0.2%) peptide. No response was observed to the irrelevant CMV pp65 peptide (0.2%) or no peptide (0.2%) controls. Further analyses were performed using MP-CTL generated from three additional HLA-A2.sup.+ donors (Donor B, Donor C, Donor D) for their CD107.alpha. degranulation or IFN-.gamma. production in response to K562-A*0201 cells presenting each individual peptide (FIG. 6b). Specific responses were detected in the MP-CTL generated from each of these donors against all the relevant peptides, but not to irrelevant CMV pp65 peptide. However, variations were detected in the level of specific response in degranulation and IFN-.gamma. production to each relevant peptide among the CTL generated from different individuals. Therefore, these studies indicate that the MP cocktail including XBP1 US, XBP1 SP, CD138, and CS1 epitopes can induce response to respective peptides with the specific CTL capable of targeting multiple antigens on MM cells.

Example 8: XBP1 Expression in Cancer Cell Lines

[0485] Various cancer cell lines were analyzed for the expression of unspliced and spliced XBP1 antigens by flow cytometry. The relative expression levels are indicated with plus or minus signs and also with numbers indicating number of plus signs (FIG. 7).

Example 9: Proliferation of XBP1-CTL in Response to HLA-A2+ Breast Cancer Cells

[0486] The proliferation of XBP1-CTL in response to stimulator cells from breast cancer cell lines was analyzed using a CFSE-proliferation assay. XBP1-CTL proliferation was measured on days 6 and 7 as a decrease in fluorescence of the CFSE-labeled MP-CTL (P3-gated cells) following stimulation with the HLA-A2.sup.+ breast cancer cell line MCF-7, HLA-A2.sup.- breast cancer cell line BT434, HLA-A2.sup.+ prostate cancer cell line LnCap, HLA-A2.sup.- prostate cancer cell line VCap and the NK-sensitive CML K562, or no cells. The XBP1-CTL showed a high level of cell proliferation in response to the HLA-A2.sup.+ breast cancer cells at days 6 (FIG. 8a) and 7 (FIG. 8b), but not other cancer cell lines which were served as controls.

Example 10: IFN-.gamma. Production and Cell Activation of XBP1-CTL in Response to HLA-A2+ Breast Cancer Cells

[0487] XBP1-CTL were analyzed by flow cytometry for IFN-.gamma. expression and CD69 upregulation in CD8+ T cells in response to breast cancer cell lines (MB231, MCF-7, BT434) and prostate cancer cell lines (LnCAP, VCap). IFN-.gamma. expression and activation (CD69) of XBP1-CTL were increased upon stimulation with HLA-A2+ breast cancer cells including MB231 and MCF-7, but not other cancer cell lines which were served as controls (FIG. 9).

Example 11: Degranulation (CD107.alpha.) of XBP1-CTL in Response to HLA-A2+ Breast Cancer Cell Lines

[0488] XBP1-CTL were analyzed for their ability to degranulate in response to breast cancer cell lines (MB231, MCF-7, BT434) and prostate cancer cell lines (LnCAP, VCap). CD107.alpha. upregulation were analyzed, as a measure of cytotoxic activity, in gated CD8.sup.+ T cells by flow cytometry. A significant level of degranulation (CD107.alpha. upregulation) was detected in response to HLA-A2+ breast cancer cells including MB231 and MCF-7, but not other cancer cell lines which were served as controls (FIG. 10).

Example 12: Proliferation of XBP1-CTL in Response to HLA-A2+ Pancreatic and Colon Cancer Cell Lines

[0489] The proliferation of XBP1-CTL in response to pancreatic and colon cancer cell lines was performed using a CFSE-based assay. XBP1-CTL proliferation was measured on day 6 as a decrease in fluorescence of the CFSE-labeled XBP1-CTL following stimulation with the HLA-A2.sup.+ prostate cancer cell line (LnCap), the HLA-A2.sup.+ pancreatic cancer cell line (8902), the HLA-A2.sup.- pancreatic cancer cell line (MiaPaca), the HLA-A2.sup.+ colon cancer cell line (LS180), and the HLA-A2.sup.- colon cancer cell line (WiDr), or a no-cell control. The XBP1-CTL showed higher levels of cell proliferation in response to the HLA-A2.sup.+ pancreatic cell line 8902 and HLA-A2.sup.+ colon cancer cell line LS180 (FIG. 11).

Example 13: IFN-.gamma. Production and Degranulation of XBP1-CTL in Response to HLA-A2+ Pancreatic and Colon Cancer Cell Lines

[0490] XBP1-CTL were analyzed for their ability to degranulate and produce IFN-.gamma. in response to pancreatic or colon cancer cell lines. XBP1-CTL were incubated with the HLA-A2+ pancreatic cancer cell lines (Pan1 and PL45), HLA-A2- pancreatic cancer cell line (MiaPaca), the HLA-A2+ colon cancer cell lines (LS180 and SW480), the HLA-A2- prostate cancer cell line (WiDr), or a no-cell control. CD107.alpha. and IFN-.gamma. production were analyzed by flow cytometry. CD107.alpha. upregulation and IFN-.gamma. production by XBP1-CTL were increased in stimulation with HLA-A2+ pancreatic cancer cells and HLA-A2+ colon cancer cells, not with other control cancer cells and showed the response in antigen-specific and HLA-A2 restricted manner (FIG. 12).

[0491] In addition, a significantly higher level of XBP1 gene expression was found in the primary tumor cells from breast cancer or colon cancer patients as compared to cells from healthy donors as shown by searching the canEvolve (http://www.canevolve.org/AnalysisResults/AnalysisResults.html) and Oncomine (http://www.webcitation.org/getfile?fileid=bcbe297e4085b19933cca- 759a88e0e2b9fac3b1e) public databases as shown below.

canEvolve

TABLE-US-00006 # Samples Exp. Status Fold Series Tumor Type Total Normal Patient p value in patients change TCGA- Colon 179 24 155 1.69E-12 Upregulated 1.60 Colon Adenocarcinoma TCGA- Breast 599 63 536 3.70E-04 Upregulated 1.64 BRCA Cancer

Oncomine

TABLE-US-00007 [0492] # Samples Exp. Status Fold Series Tumor Type Total Normal Patient p value in patients change TCGA- Colon 161 Colon Adenocarcinoma #1. Mutinous 19 22 1.92E-06 Upregulated 1.91 Adenocarcinoma #2. Adenocarcinoma 19 101 0.016 Upregulated 1.23 TCGA- Breast 593 BRCA cancer #1. Invasive lobular 61 36 3.45E-07 Upregulated 2.63 #2. Invasive Ductal 61 392 6.19E-05 Upregulated 1.77 #3. Mixed lobular 61 7 1.67E-04 Upregulated 3.67 and Ductal #4. Intraductal 61 3 2.86E-04 Upregulated 2.71 Cribriform #5. Invasive Ductal 61 3 0.003 Upregulated 3.04 & Lobular #6. Invasive 61 76 0.007 Upregulated 1.54 carcinoma

Example 14: CD138 and CS1 Expression in Cancer Cell Lines

[0493] Various cancer cell lines were analyzed for CD138 and CS1 antigen expression by flow cytometry. The relative CD138 and CS-1 expression levels for the various cancer cell lines are indicated with plus or minus signs and also with numbers indicating the expression levels of CD138 (FIG. 13) and CS1 (FIG. 14).

Example 15: Percentage of CD8+ CTL in T Cells from Smoldering Multiple Myeloma Patients by Stimulation with XBP1/CD138/CS1 Peptides

[0494] Multipeptide-specific CTL (MP-CTL) generated from T cells obtained from four HLA-A2+ smoldering multiple myeloma patients were analyzed for specific T cells by flow cytometry. The MP-CTL generated from each patient's T cells show an increased percentage of CD3+CD8+ CTLs and decreased percentage of CD3+CD4+Th cells (FIG. 15).

Example 16: Proliferation of MP-CTL from Smoldering Multiple Myeloma Patients

[0495] Proliferation of MP-CTL generated from two smoldering multiple myeloma patients in response to various MM cell lines was analyzed using a CFSE-based assay. Cell proliferation was measured on days 5, 6 and 7 as a decrease in fluorescence of the CFSE-labeled MP-CTL (P3-gated) in stimulation with the HLA-A2.sup.+ cell line (McCAR), the HLA-A2.sup.- cell lines (MM15, RPMI), NK sensitive cell line (K562), or without stimulation. MP-CTL generated from T cells of both SMM patients showed a high level of cell proliferation in response to the HLA-A2.sup.+ multiple myeloma cell line McCAR, not to HLA-A2- cell lines nor NK sensitive cell line, demonstrating HLA-A2 restricted MP-CTL activities against multiple myeloma cells (FIGS. 16a, b).

Example 17: IFN-.gamma. Production by MP-CTL Generated from Smoldering Multiple Myeloma Patients

[0496] IFN-.gamma. production of MP-CTL generated from four smoldering multiple myeloma patients was analyzed in response to various MM cell lines. The production was analyzed by flow cytometry following stimulation with an HLA-A2+ cell line (McCAR, U266), the HLA-A2- cell lines (MM1S, RPMI), the NK sensitive cell line (K562) or without any stimulation. The MP-CTL generated from all four patients showed an increase in IFN-.gamma. production in an HLA-A2-restricted manner upon the stimulation with HLA-A2.sup.+ multiple myeloma cell lines (FIGS. 17a, b).

Example 18: Degranulation (CD107.alpha.) of MP-CTL Generated from Smoldering Multiple Myeloma Patients to Myeloma Cell Lines

[0497] MP-CTL generated from four smolder multiple myeloma patients were analyzed for degranulation as a measure of cytotoxic activity in response to various HLA-A2+ and HLA-A2-cancer cell lines. The analyses were performed by measuring the MP-CTL specific responses to U266, McCAR, MM1S, RPMI, K562 cells, or no stimulator cells. CD107.alpha. upregulation was analyzed in gated CD8+ T cells by flow cytometry. The MP-CTL generated from a SMM patient showed an increased level of CD107.alpha. degranulation in response to HLA-A2+MM cell lines, U266 and McCAR (FIG. 18a). The HLA-A2 restricted degranulation response to multiple myeloma cells was confirmed using the MP-CTL generated from four SMM patients (FIG. 18b).

Example 19: Percentage of IFN-.gamma.+CD107.alpha.+ Double Positive Cells Among the CD3+CD8+CD137+ Subset from MP-CTL Generated from Smoldering Multiple Myeloma Patients

[0498] The CD3+CD8+CD137+ subset of MP-CTL generated from smoldering multiple myeloma patients were analyzed for their ability to degranulate (CD107.alpha. expression) and for IFN-.gamma. production in response to K562-A2 cells presenting individual peptides. CD107.alpha. and/or IFN-.gamma. expression was analyzed within the CD3+CD8+CD137+ subset using flow cytometry. MP-CTL generated from three SMM patients demonstrated increased levels of peptide-specific IFN-.gamma. production and/or CD107.alpha. degranulation (FIGS. 19a-d).

Example 20: Phenotypic Characterization of MP-CTL Generated from Smoldering Multiple Myeloma Patients

[0499] MP-CTL generated by a total four multipeptide stimulation were analyzed for native, central memory (CM), effector memory (EM) and terminal effector (TE) CD8+T subsets by flow cytometry. MP-CTL generated from four SMM patients show an increased percentage of EM and TE CD8.sup.+ cells (FIG. 20a). The MP-CTL from SMM patient #2 and SMM patient #4 were received an additional three rounds of multipeptide stimulation (a total seven cycles of stimulation) and were analyzed by flow cytometry. Additional three cycles of MP stimulation resulted in a further expansion of the EM subset (cycle 4 vs. cycle 7) (FIG. 20b). In further experiments, the EM and TE subsets were evaluated from the MP-CTL generated from three SMM patients for IFN-.gamma. production and CD107.alpha. upregulation in response to various cancer cell lines. Compared to TE, EM cells within the MP-CTL from all three patients show a higher percentage of IFN-.gamma.+CD107.alpha.+ double positive cell in response to HLA-A2+MM cell line McCAR (FIG. 20c). In addition, compared to native (CD45RO.sup.-CCR7.sup.+) CD8.sup.+ T cells, memory (CD45RO.sup.+) type of CD8.sup.+ T cells showed a higher expression of CD69 activation marker within the MP-CTL generated from four smoldering multiple myeloma patients against HLA-A2.sup.+ multiple myeloma cell line 11266, confirming higher anti-tumor activities of MP-CTL by memory subsets (FIG. 20d).

Example 21. Peptides from Unspliced XBP1, Spliced XBP1, CD138, and CS1 Bind HLA-A24 with High Affinity

[0500] The full length sequences of non-spliced or spliced XBP1 protein (see above) were analyzed using the search software SYFPEITHI (A database of MHC ligands and Peptide Motifs, Institute for Cell Biology, Department of Immunology, Heidlberg) to predict peptides specific to HLA-A24 followed by the BIMAS program to select peptides with extended half-time disassociation rates. The following peptides from non-spliced XBP1 were selected as potential HLA-A24-binding peptides: peptide 1 (SEQ ID NO: 33), peptide 2 (SEQ ID NO: 5), peptide 3 (SEQ ID NO: 34), peptide 4 (SEQ ID NO: 35), peptide 5 (SEQ ID NO: 36), peptide 6 (SEQ ID NO: 37), and peptide 7 (SEQ ID NO: 29). The following peptides from spliced XBP1 were selected as potential HLA-A24-binding peptides: peptide 1 (SEQ ID NO: 30), peptide 2 (SEQ ID NO: 38), peptide 3 (SEQ ID NO: 39), and peptide 4 (SEQ ID NO: 5). The following peptides from CD138 were selected as potential HLA-A24-binding peptides: peptide 1 (SEQ ID NO: 31), peptide 2 (SEQ ID NO: 40), peptide 3 (SEQ ID NO: 41), peptide 4 (SEQ ID NO: 42), peptide 5 (SEQ ID NO: 43), peptide 6 (SEQ ID NO: 44), and peptide 7 (SEQ ID NO: 45). The following peptides from CS1 were selected as potential HLA-A24-binding peptides: peptide 1 (SEQ ID NO: 46), peptide 2 (SEQ ID NO: 47), peptide 3 (SEQ ID NO: 48), peptide 4 (SEQ ID NO: 49), peptide 5 (SEQ ID NO: 32), and peptide 6 (SEQ ID NO: 50). The HLA-A24 affinity of these native XBP1 peptides and HIV envelope protein.sub.583-591 (SEQ ID NO: 537), which is known to be a HLA-A24-binding peptide, was assessed using the T2 peptide-binding assay at a peptide concentration of 1 mg/ml. The specific affinity of the peptides was evaluated as HLA-A24-mean fluorescence intensity (MFI), which is a function of HLA-A24 upregulation on T2 cells following peptide binding to HLA-A24. Among the tested peptides, the following peptides displayed binding affinities for HLA-A24 comparable to or higher than that of HIV envelope protein.sub.583-591 (SEQ ID NO: 537): peptides 4 and 7 from non-spliced XBP1, peptide 1 from spliced XBP1, peptides 1, 3, and 4 from CD138, and peptides 3 and 5 from CS1 (FIG. 21). The HLA-A24 affinities of the high-affinity peptides were then analyzed in the T2 peptide-binding assay using lower peptide concentrations (FIGS. 22-24). Based on these data, the following four peptides, including one peptide from each protein, were selected for further investigation: peptide 7 from non-spliced XBP1 (XBP1.sub.186-194, sequence: ISPWILAVL, SEQ ID NO: 29), peptide 1 from spliced XBP1 (SPXBP1.sub.224-232, sequence: VYPEGPSSL, SEQ ID NO: 30), peptide 3 from CD138 (CD138.sub.265-273, sequence: IFAVCLVGF, SEQ ID NO: 31), and peptide 5 from CS1 (CS.sub.1240-248, sequence: LFVLGLFLW, SEQ ID NO: 32).

Example 22: Percentage of CD8+ CTL in T Cells from Two Donors by Stimulation with XBP1, CD138, and CS1 Peptides

[0501] CTL from two donor patients were analyzed for specific T cells by flow cytometry. The peptide-specific CTL generated from each patient's T cells show an increased percentage of CD8+ CTL (FIG. 26).

Example 23: IFN-.gamma. Production and Cell Activation of Peptide-Specific CTL in Response to HLA-A24+ Multiple Myeloma Cells

[0502] Peptide-specific CTL were analyzed by flow cytometry for expression of IFN-.gamma. and CD8 in response to HLA-A24+(KMS) and HLA-A24-(OMP1 and 11266) multiple myeloma cell lines. IFN-.gamma. expression of peptide-specific CTL was increased upon stimulation with HLA-A24+ but not with HLA-A24-multiple myeloma cells (FIGS. 28 and 29).

Example 24: IFN-.gamma. Production and Degranulation of Peptide-Specific CTL in Response to HLA-A24+ Multiple Myeloma Cell Lines

[0503] Peptide-specific CTL were analyzed for their ability to degranulate and produce IFN-.gamma. in response to multiple myeloma cell lines. Peptide-specific CTL were incubated with the HLA-A24+(KMS11) and HLA-A24-(OMP1) multiple myeloma cell lines. CD107.alpha. and IFN-.gamma. production were analyzed by flow cytometry. CD107.alpha. upregulation and IFN-.gamma. production by peptide-specific CTL were increased by stimulation with HLA-A24+ but not with HLA-A24-multiple myeloma cells (FIG. 31).

Example 25: Proliferation of Peptide-Specific CTL in Response to HLA-A24+ Multiple Myeloma Cells

[0504] The proliferation of peptide-specific CTL in response to stimulation from multiple myeloma cells was analyzed using a CFSE-proliferation assay. Peptide-specific CTL proliferation was measured on days 6 and 8 as a decrease in fluorescence of the CFSE-labeled MP-CTL (P3-gated cells) following either stimulation with the HLA-A24.sup.+ multiple myeloma cell line KMS11 or the HLA-A24.sup.- multiple myeloma cell line OMP1 or no stimulation. The peptide-specific CTL showed a high level of cell proliferation at days 6 (FIG. 33a) and 8 (FIG. 33b) in response to the HLA-A24.sup.+ multiple myeloma cells but not the HLA-A24.sup.- multiple myeloma cells.

Example 26: IL-2 Production by Peptide-Specific CTL in Response to HLA-A24+ Multiple Myeloma Cells

[0505] IL-2 production by peptide-specific CTL in response to stimulation from multiple myeloma cells was analyzed by flow cytometry. IL-2 production by peptide-specific CTL proliferation was measured following either stimulation with the HLA-A24.sup.+ multiple myeloma cell line KMS11 or the HLA-A24.sup.- multiple myeloma cell lines OMP1 and U266 or no stimulation. The peptide-specific CTL showed a high level of IL-2 production in response to the HLA-A24.sup.+ multiple myeloma cells but not the HLA-A24.sup.- multiple myeloma cells (FIG. 34).

Example 27: IFN-.gamma. Production and Cell Activation of Peptide-Specific CTL in Response to HLA-A24+ Colon Cancer Cells

[0506] Peptide-specific CTL from two donors were analyzed by flow cytometry for expression of IFN-.gamma. and CD8 in response to HLA-A24+(SW480) and HLA-A24-(WiDr and LS180) colon cancer cell lines. CD8 and IFN-.gamma. expression were analyzed by flow cytometry. IFN-.gamma. expression of peptide-specific CTL was increased upon stimulation with HLA-A24+ but not with HLA-A24-colon cancer cells (FIGS. 35 and 37).

Example 28: IFN-.gamma. Production and Degranulation of Peptide-Specific CTL in Response to HLA-A24+ Colon Cancer Cells

[0507] Peptide-specific CTL were analyzed for their ability to degranulate and produce IFN-.gamma. in response to HLA-A24+(SW480) and HLA-A24-(WiDr and LS180) colon cancer cell lines. CD107.alpha. and IFN-.gamma. production were analyzed by flow cytometry. CD107.alpha. upregulation and IFN-.gamma. production by peptide-specific CTL were increased by stimulation with HLA-A24+ but not with HLA-A24-colon cancer cells (FIGS. 36 and 38).

Example 29: Degranulation and Production of IFN-.gamma. and IL-2 in CD138-CTL in Response to Varying Amounts of HLA-A24+ Colon Cancer Cells

[0508] CD138-CTL were analyzed for their ability to degranulate and produce IFN-.gamma. and IL-2 in response to HLA-A24+(SW480) and HLA-A24-(LS180) colon cancer cells at 5:1, 1:1, and 1:5 ratios of CTL to tumor cells. Degranulation, IFN-.gamma. production, and IL-2 production displayed a substantial increase at higher ratios of CTL to tumor cells upon stimulation with HLA-A24+ colon cancer cells (FIG. 39a) but not with HLA-A24-colon cancer cells (FIG. 39b).

Example 30: Cell Activation, IFN-.gamma. Production and Degranulation of HLA-A24-Specific CD138 Peptide-Specific CTL in Response to HLA-A24+ Colon Cancer and Multiple Myeloma Cells

[0509] Peptide-specific CTL from two donors were analyzed for CD8 expression, degranulation, and produce IFN-.gamma. production in response to HLA-A24+ colon cancer (SW480) and multiple myeloma (KMS11) cell lines. Spliced XBP1-CTL and CD138-CTL from both donors showed increased activation, IFN-.gamma. production, and degranulation in response to colon cancer cells (FIGS. 40 and 41). Unspliced XBP1-CTL from Donor A showed increased activation, IFN-.gamma. production, and degranulation in response to colon cancer cells, whereas these effects were stronger in response to multiple myeloma cells in unspliced XBP1-CTL from the Donor B (FIGS. 40 and 41).

Example 31: IFN-.gamma. Production and Cell Activation of Peptide-Specific CD138-CTL in Response to HLA-A24+ Pancreatic Cancer Cells

[0510] Peptide-specific CTL from Donor B were analyzed by flow cytometry for expression of IFN-.gamma. and CD8 in response to HLA-A24+(8902 and PL45) and HLA-A24-(MiaPaca) colon cancer cell lines. CD8 and IFN-.gamma. expression were analyzed by flow cytometry. IFN-.gamma. expression of peptide-specific CTL was increased upon stimulation with HLA-A24+ but not with HLA-A24-pancreatic cancer cells (FIG. 42).

Example 32: Degranulation and Production of IFN-.gamma. and IL-2 in CD138-CTL in Response to Varying Amounts of HLA-A24+ Pancreatic Cancer Cells

[0511] CD138-CTL were analyzed for their ability to degranulate and produce IFN-.gamma. and IL-2 in response to HLA-A24+(Panel) pancreatic cancer cells at 1:1 and 1:5 ratios of CTL to tumor cells. Degranulation, IFN-.gamma. production, and IL-2 production were higher at a 1:1 ratio of CTL to tumor cells (FIG. 43).

Example 33: Induction of Functionally Active Memory Cells with Heteroclitic Unspliced XBP1.sub.184-192 (YISPWILAV) (SEQ ID NO: 6) and Heteroclitic Spliced XBP1 SP.sub.196-204 (YLFPQLISV) (SEQ ID NO: 10) Peptides

[0512] Cytotoxic T lymphocytes (CTL) induced with a cocktail of heteroclitic unspliced XBP1.sub.184-192 (YISPWILAV) (SEQ ID NO: 6) and heteroclitic spliced XBP1 SP.sub.196-204 (YLFPQLISV) (SEQ ID NO: 10) peptides contain both central memory (CM) and effector memory (EM) CD8.sup.+ T cells (FIGS. 44, 45 and 46), which can recognize and target the specific XBP1 antigens on tumor cells. XBP1 peptides-specific CTL (XBP1-CTL) demonstrate a high level of cell proliferation, within the memory CTL subsets in response to breast (MB231; FIG. 47), colon (LS180; FIG. 48) or pancreatic (Panc1; FIG. 49) cancer cell lines. Overall, effector memory CTL had a higher level of cell proliferation as compared to central memory CTL (FIGS. 47, 48 and 49) in response to the different tumor cell lines. In contrast, central memory CTL induced a higher level of IFN-.gamma. production, IL-2 production and cytotoxic activities as compared to effector memory CTL in response to HLA-A2.sup.+ breast (MB231, MCF7), colon (LS180, SW480) and pancreatic (Panc1, PL45) cancer cell lines (FIGS. 50, 51, 52, 53, 54, 55). T-bet and Eomes are critical transcriptional regulators found in effector and memory T cells, which have roles relating to anti-tumor activities. XBP1-CTL memory cells displayed a high level of both T-bet (FIGS. 56, 57, 58) and Eomes expression (FIGS. 62, 63, 64). In response to breast, colon and pancreatic cancer lines, the T-bet.sup.+ (FIGS. 59, 60, 61) and Eome.sup.+ (FIGS. 65, 66, 67) cells display a high level of IFN-.gamma. production. Memory cells within the XBP1-CTL were evaluated for upregulation of Granzyme B (indicator of anti-tumor cytotoxic activity) against breast, colon and pancreatic cancer lines. Compared to non-memory cells, memory cells showed a higher level of granzyme upregulation (FIG. 68). In addition, the granzyme.sup.+ XPB1-CTL also produced IFN-.gamma. in response to breast cancer, colon or pancreatic cancer cells (FIGS. 69, 70, 71). Therefore, the CTL induced with a cocktail of heteroclitic unspliced XBP1.sub.184-192 (YISPWILAV) (SEQ ID NO: 6) and heteroclitic spliced XBP1 SP.sub.196-204 (YLFPQLISV) (SEQ ID NO: 10) peptides contain both central memory and effector memory CD8.sup.+ T cells, and those memory cells of the XBP1-CTL showed anti-tumor activities against breast cancer, colon cancer and pancreatic cancer cells.

Example 34: Lenalidomide Treatment Increases the Total Number and Function of Memory Cell Subsets within XBP1-CTL

[0513] The number of memory CD8+ T cells is increased within the XBP1 peptide-specific CTL following treatment with Lenalidomide (at 5 uM for 5 days) (FIG. 72). This observation corresponds with a decrease in total non-memory CD8.sup.+ T cells within the XBP1-CTL. A consistent increase or some increase of the frequency of central memory CD8.sup.+ T cells or central memory CD3.sup.+CD8.sup.+ T cells, respectively, is detected after treatment with Lenalidomide (FIG. 73). In addition, critical markers for T cell activation and other immunological functions (i.e., CD40L, CD69, CD38) are increased on XBP1-CTL upon treatment with Lenalidomide (FIG. 74). The total CD3.sup.+CD8.sup.+ T cells, central memory CD3.sup.+CD8.sup.+ T cells and effector memory CD3.sup.+CD8.sup.+ T cells of XBP1-CTL display increased levels of T-bet expression/IFN-7 production and Eomes expression/IFN-g production in response to breast (MB231; FIGS. 75, 76), pancreatic (Panc1; FIG. 77, 78) or colon (SW480; FIGS. 79, 80) cancer cells by treatment with Lenalidomide. Memory XBP1-CTL treated with Lenalidomide have increased polyfunctional immune responses (cytotoxicity and IFN-.gamma. production) against breast (MB231; FIG. 81), pancreatic (Panc1; FIG. 82) or colon (SW480; FIG. 83) cancer cells as compared to the XBP1 CTL alone. Therefore, these results provide evidence that anti-activities of CTL induced with a cocktail of heteroclitic unspliced XBP1.sub.184-192 (YISPWILAV) (SEQ ID NO: 6) and heteroclitic spliced XBP1SP.sub.196-204 (YLFPQLISV) (SEQ ID NO: 10) peptides were enhanced by the treatment of CTL with Lenalidomide.

OTHER EMBODIMENTS

[0514] While the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Sequence CWU 1

1

54419PRTHomo sapiens 1Leu Leu Arg Glu Lys Thr His Gly Leu 1 5 29PRTHomo sapiens 2Asn Ile Ser Pro Trp Ile Leu Ala Val 1 5 39PRTHomo sapiens 3Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 49PRTHomo sapiens 4Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 59PRTHomo sapiens 5Lys Leu Leu Leu Glu Asn Gln Leu Leu 1 5 69PRTHomo sapiens 6Tyr Ile Ser Pro Trp Ile Leu Ala Val 1 5 79PRTHomo sapiens 7Gly Ile Leu Asp Asn Leu Asp Pro Val 1 5 89PRTHomo sapiens 8Ile Leu Leu Gly Ile Leu Asp Asn Leu 1 5 99PRTHomo sapiens 9Glu Leu Phe Pro Gln Leu Ile Ser Val 1 5 109PRTHomo sapiens 10Tyr Leu Phe Pro Gln Leu Ile Ser Val 1 5 119PRTHomo sapiens 11Val Ile Ala Gly Gly Leu Val Gly Leu 1 5 129PRTHomo sapiens 12Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 139PRTHomo sapiens 13Ala Leu Trp Leu Trp Leu Cys Ala Leu 1 5 149PRTHomo sapiens 14Trp Leu Trp Leu Cys Ala Leu Ala Leu 1 5 1510PRTHomo sapiens 15Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 169PRTHomo sapiens 16Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 179PRTHomo sapiens 17Leu Leu Val Pro Leu Leu Leu Ser Leu 1 5 189PRTHomo sapiens 18Thr Leu Ile Tyr Ile Leu Trp Gln Leu 1 5 19261PRTHomo sapiens 19Met Val Val Val Ala Ala Ala Pro Asn Pro Ala Asp Gly Thr Pro Lys 1 5 10 15 Val Leu Leu Leu Ser Gly Gln Pro Ala Ser Ala Ala Gly Ala Pro Ala 20 25 30 Gly Gln Ala Leu Pro Leu Met Val Pro Ala Gln Arg Gly Ala Ser Pro 35 40 45 Glu Ala Ala Ser Gly Gly Leu Pro Gln Ala Arg Lys Arg Gln Arg Leu 50 55 60 Thr His Leu Ser Pro Glu Glu Lys Ala Leu Arg Arg Lys Leu Lys Asn 65 70 75 80 Arg Val Ala Ala Gln Thr Ala Arg Asp Arg Lys Lys Ala Arg Met Ser 85 90 95 Glu Leu Glu Gln Gln Val Val Asp Leu Glu Glu Glu Asn Gln Lys Leu 100 105 110 Leu Leu Glu Asn Gln Leu Leu Arg Glu Lys Thr His Gly Leu Val Val 115 120 125 Glu Asn Gln Glu Leu Arg Gln Arg Leu Gly Met Asp Ala Leu Val Ala 130 135 140 Glu Glu Glu Ala Glu Ala Lys Gly Asn Glu Val Arg Pro Val Ala Gly 145 150 155 160 Ser Ala Glu Ser Ala Ala Leu Arg Leu Arg Ala Pro Leu Gln Gln Val 165 170 175 Gln Ala Gln Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala 180 185 190 Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser Cys Trp Ala Phe Trp 195 200 205 Thr Thr Trp Thr Gln Ser Cys Ser Ser Asn Ala Leu Pro Gln Ser Leu 210 215 220 Pro Ala Trp Arg Ser Ser Gln Arg Ser Thr Gln Lys Asp Pro Val Pro 225 230 235 240 Tyr Gln Pro Pro Phe Leu Cys Gln Trp Gly Arg His Gln Pro Ser Trp 245 250 255 Lys Pro Leu Met Asn 260 20376PRTHomo sapiens 20Met Val Val Val Ala Ala Ala Pro Asn Pro Ala Asp Gly Thr Pro Lys 1 5 10 15 Val Leu Leu Leu Ser Gly Gln Pro Ala Ser Ala Ala Gly Ala Pro Ala 20 25 30 Gly Gln Ala Leu Pro Leu Met Val Pro Ala Gln Arg Gly Ala Ser Pro 35 40 45 Glu Ala Ala Ser Gly Gly Leu Pro Gln Ala Arg Lys Arg Gln Arg Leu 50 55 60 Thr His Leu Ser Pro Glu Glu Lys Ala Leu Arg Arg Lys Leu Lys Asn 65 70 75 80 Arg Val Ala Ala Gln Thr Ala Arg Asp Arg Lys Lys Ala Arg Met Ser 85 90 95 Glu Leu Glu Gln Gln Val Val Asp Leu Glu Glu Glu Asn Gln Lys Leu 100 105 110 Leu Leu Glu Asn Gln Leu Leu Arg Glu Lys Thr His Gly Leu Val Val 115 120 125 Glu Asn Gln Glu Leu Arg Gln Arg Leu Gly Met Asp Ala Leu Val Ala 130 135 140 Glu Glu Glu Ala Glu Ala Lys Gly Asn Glu Val Arg Pro Val Ala Gly 145 150 155 160 Ser Ala Glu Ser Ala Ala Gly Ala Gly Pro Val Val Thr Pro Pro Glu 165 170 175 His Leu Pro Met Asp Ser Gly Gly Ile Asp Ser Ser Asp Ser Glu Ser 180 185 190 Asp Ile Leu Leu Gly Ile Leu Asp Asn Leu Asp Pro Val Met Phe Phe 195 200 205 Lys Cys Pro Ser Pro Glu Pro Ala Ser Leu Glu Glu Leu Pro Glu Val 210 215 220 Tyr Pro Glu Gly Pro Ser Ser Leu Pro Ala Ser Leu Ser Leu Ser Val 225 230 235 240 Gly Thr Ser Ser Ala Lys Leu Glu Ala Ile Asn Glu Leu Ile Arg Phe 245 250 255 Asp His Ile Tyr Thr Lys Pro Leu Val Leu Glu Ile Pro Ser Glu Thr 260 265 270 Glu Ser Gln Ala Asn Val Val Val Lys Ile Glu Glu Ala Pro Leu Ser 275 280 285 Pro Ser Glu Asn Asp His Pro Glu Phe Ile Val Ser Val Lys Glu Glu 290 295 300 Pro Val Glu Asp Asp Leu Val Pro Glu Leu Gly Ile Ser Asn Leu Leu 305 310 315 320 Ser Ser Ser His Cys Pro Lys Pro Ser Ser Cys Leu Leu Asp Ala Tyr 325 330 335 Ser Asp Cys Gly Tyr Gly Gly Ser Leu Ser Pro Phe Ser Asp Met Ser 340 345 350 Ser Leu Leu Gly Val Asn His Ser Trp Glu Asp Thr Phe Ala Asn Glu 355 360 365 Leu Phe Pro Gln Leu Ile Ser Val 370 375 21310PRTHomo sapiens 21Met Arg Arg Ala Ala Leu Trp Leu Trp Leu Cys Ala Leu Ala Leu Ser 1 5 10 15 Leu Gln Pro Ala Leu Pro Gln Ile Val Ala Thr Asn Leu Pro Pro Glu 20 25 30 Asp Gln Asp Gly Ser Gly Asp Asp Ser Asp Asn Phe Ser Gly Ser Gly 35 40 45 Ala Gly Ala Leu Gln Asp Ile Thr Leu Ser Gln Gln Thr Pro Ser Thr 50 55 60 Trp Lys Asp Thr Gln Leu Leu Thr Ala Ile Pro Thr Ser Pro Glu Pro 65 70 75 80 Thr Gly Leu Glu Ala Thr Ala Ala Ser Thr Ser Thr Leu Pro Ala Gly 85 90 95 Glu Gly Pro Lys Glu Gly Glu Ala Val Val Leu Pro Glu Val Glu Pro 100 105 110 Gly Leu Thr Ala Arg Glu Gln Glu Ala Thr Pro Arg Pro Arg Glu Thr 115 120 125 Thr Gln Leu Pro Thr Thr His Gln Ala Ser Thr Thr Thr Ala Thr Thr 130 135 140 Ala Gln Glu Pro Ala Thr Ser His Pro His Arg Asp Met Gln Pro Gly 145 150 155 160 His His Glu Thr Ser Thr Pro Ala Gly Pro Ser Gln Ala Asp Leu His 165 170 175 Thr Pro His Thr Glu Asp Gly Gly Pro Ser Ala Thr Glu Arg Ala Ala 180 185 190 Glu Asp Gly Ala Ser Ser Gln Leu Pro Ala Ala Glu Gly Ser Gly Glu 195 200 205 Gln Asp Phe Thr Phe Glu Thr Ser Gly Glu Asn Thr Ala Val Val Ala 210 215 220 Val Glu Pro Asp Arg Arg Asn Gln Ser Pro Val Asp Gln Gly Ala Thr 225 230 235 240 Gly Ala Ser Gln Gly Leu Leu Asp Arg Lys Glu Val Leu Gly Gly Val 245 250 255 Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 260 265 270 Phe Met Leu Tyr Arg Met Lys Lys Lys Asp Glu Gly Ser Tyr Ser Leu 275 280 285 Glu Glu Pro Lys Gln Ala Asn Gly Gly Ala Tyr Gln Lys Pro Thr Lys 290 295 300 Gln Glu Glu Phe Tyr Ala 305 310 22335PRTHomo sapiens 22Met Ala Gly Ser Pro Thr Cys Leu Thr Leu Ile Tyr Ile Leu Trp Gln 1 5 10 15 Leu Thr Gly Ser Ala Ala Ser Gly Pro Val Lys Glu Leu Val Gly Ser 20 25 30 Val Gly Gly Ala Val Thr Phe Pro Leu Lys Ser Lys Val Lys Gln Val 35 40 45 Asp Ser Ile Val Trp Thr Phe Asn Thr Thr Pro Leu Val Thr Ile Gln 50 55 60 Pro Glu Gly Gly Thr Ile Ile Val Thr Gln Asn Arg Asn Arg Glu Arg 65 70 75 80 Val Asp Phe Pro Asp Gly Gly Tyr Ser Leu Lys Leu Ser Lys Leu Lys 85 90 95 Lys Asn Asp Ser Gly Ile Tyr Tyr Val Gly Ile Tyr Ser Ser Ser Leu 100 105 110 Gln Gln Pro Ser Thr Gln Glu Tyr Val Leu His Val Tyr Glu His Leu 115 120 125 Ser Lys Pro Lys Val Thr Met Gly Leu Gln Ser Asn Lys Asn Gly Thr 130 135 140 Cys Val Thr Asn Leu Thr Cys Cys Met Glu His Gly Glu Glu Asp Val 145 150 155 160 Ile Tyr Thr Trp Lys Ala Leu Gly Gln Ala Ala Asn Glu Ser His Asn 165 170 175 Gly Ser Ile Leu Pro Ile Ser Trp Arg Trp Gly Glu Ser Asp Met Thr 180 185 190 Phe Ile Cys Val Ala Arg Asn Pro Val Ser Arg Asn Phe Ser Ser Pro 195 200 205 Ile Leu Ala Arg Lys Leu Cys Glu Gly Ala Ala Asp Asp Pro Asp Ser 210 215 220 Ser Met Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu 225 230 235 240 Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg Glu Arg Gln Glu 245 250 255 Glu Tyr Ile Glu Glu Lys Lys Arg Val Asp Ile Cys Arg Glu Thr Pro 260 265 270 Asn Ile Cys Pro His Ser Gly Glu Asn Thr Glu Tyr Asp Thr Ile Pro 275 280 285 His Thr Asn Arg Thr Ile Leu Lys Glu Asp Pro Ala Asn Thr Val Tyr 290 295 300 Ser Thr Val Glu Ile Pro Lys Lys Met Glu Asn Pro His Ser Leu Leu 305 310 315 320 Thr Met Pro Asp Thr Pro Arg Leu Phe Ala Tyr Glu Asn Val Ile 325 330 335 234PRTUnknownDescription of Unknown "KDEL" motif peptide 23Lys Asp Glu Leu 1 249PRTHomo sapiens 24Tyr Ile Leu Asp Asn Leu Asp Pro Val 1 5 259PRTInfluenza virus 25Gly Ile Leu Gly Phe Val Phe Thr Leu 1 5 269PRTInfluenza virus 26Phe Leu Trp Gly Pro Arg Ala Leu Val 1 5 279PRTHomo sapiens 27Arg Met Ser Glu Leu Glu Gln Gln Val 1 5 289PRTHomo sapiens 28Asn Leu Val Pro Met Val Ala Thr Val 1 5 299PRTHomo sapiens 29Ile Ser Pro Trp Ile Leu Ala Val Leu 1 5 309PRTHomo sapiens 30Val Tyr Pro Glu Gly Pro Ser Ser Leu 1 5 319PRTHomo sapiens 31Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 329PRTHomo sapiens 32Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 339PRTHomo sapiens 33Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 349PRTHomo sapiens 34Glu Asn Gln Glu Leu Arg Gln Arg Leu 1 5 359PRTHomo sapiens 35Asp Gly Thr Pro Lys Val Leu Leu Leu 1 5 369PRTHomo sapiens 36Asp Leu Glu Glu Glu Asn Gln Lys Leu 1 5 379PRTHomo sapiens 37Ser Ser Asn Ala Leu Pro Gln Ser Leu 1 5 389PRTHomo sapiens 38Gly Tyr Gly Gly Ser Leu Ser Pro Phe 1 5 399PRTHomo sapiens 39Pro Phe Ser Asp Met Ser Ser Leu Leu 1 5 409PRTHomo sapiens 40Val Leu Pro Glu Val Glu Pro Gly Leu 1 5 419PRTHomo sapiens 41Leu Pro Gln Ile Val Ala Thr Asn Leu 1 5 429PRTHomo sapiens 42Leu Ala Leu Ser Leu Gln Pro Ala Leu 1 5 439PRTHomo sapiens 43Gly Leu Leu Asp Arg Lys Glu Val Leu 1 5 449PRTHomo sapiens 44Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 459PRTHomo sapiens 45Ser Leu Gln Pro Ala Leu Pro Gln Ile 1 5 469PRTHomo sapiens 46Gly Tyr Ser Leu Lys Leu Ser Lys Leu 1 5 479PRTHomo sapiens 47Asp Phe Pro Asp Gly Gly Tyr Ser Leu 1 5 489PRTHomo sapiens 48Thr Met Pro Asp Thr Pro Arg Leu Phe 1 5 499PRTHomo sapiens 49Arg Trp Gly Glu Ser Asp Met Thr Phe 1 5 509PRTHomo sapiens 50Lys Met Glu Asn Pro His Ser Leu Leu 1 5 5115PRTHomo sapiens 51Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu 1 5 10 15 5216PRTHomo sapiens 52Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 5317PRTHomo sapiens 53Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu 5418PRTHomo sapiens 54Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu Gln 5519PRTHomo sapiens 55Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu Gln Ile 5620PRTHomo sapiens 56Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu Gln Ile Gln 20 5721PRTHomo sapiens 57Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu Gln Ile Gln Ser 20 5822PRTHomo sapiens 58Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu Gln Ile Gln Ser Leu 20 5923PRTHomo sapiens 59Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu Gln Ile Gln Ser Leu Ile 20 6024PRTHomo sapiens 60Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu Gln Ile Gln Ser Leu Ile Ser 20 6125PRTHomo sapiens 61Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu Gln Ile Gln Ser Leu Ile Ser Cys 20 25 6226PRTHomo sapiens 62Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu Gln Ile Gln Ser Leu Ile Ser Cys Trp 20 25 6327PRTHomo sapiens 63Leu Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 Leu Gln Ile Gln Ser Leu Ile Ser Cys Trp Ala 20 25 6414PRTHomo sapiens 64Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu 1 5 10 6515PRTHomo sapiens 65Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 15 6616PRTHomo sapiens 66Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 6717PRTHomo sapiens 67Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 Gln 6818PRTHomo sapiens 68Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 Gln Ile 6919PRTHomo sapiens 69Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 Gln Ile Gln 7020PRTHomo sapiens 70Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 Gln Ile Gln Ser 20 7121PRTHomo sapiens 71Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 Gln Ile Gln Ser Leu 20

7222PRTHomo sapiens 72Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 Gln Ile Gln Ser Leu Ile 20 7323PRTHomo sapiens 73Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 Gln Ile Gln Ser Leu Ile Ser 20 7424PRTHomo sapiens 74Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 Gln Ile Gln Ser Leu Ile Ser Cys 20 7525PRTHomo sapiens 75Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 Gln Ile Gln Ser Leu Ile Ser Cys Trp 20 25 7626PRTHomo sapiens 76Ser Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 Gln Ile Gln Ser Leu Ile Ser Cys Trp Ala 20 25 7713PRTHomo sapiens 77Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu 1 5 10 7814PRTHomo sapiens 78Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 7915PRTHomo sapiens 79Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15 8016PRTHomo sapiens 80Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 8117PRTHomo sapiens 81Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 Ile 8218PRTHomo sapiens 82Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 Ile Gln 8319PRTHomo sapiens 83Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 Ile Gln Ser 8420PRTHomo sapiens 84Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 Ile Gln Ser Leu 20 8521PRTHomo sapiens 85Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 Ile Gln Ser Leu Ile 20 8622PRTHomo sapiens 86Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 Ile Gln Ser Leu Ile Ser 20 8723PRTHomo sapiens 87Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 Ile Gln Ser Leu Ile Ser Cys 20 8824PRTHomo sapiens 88Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 Ile Gln Ser Leu Ile Ser Cys Trp 20 8925PRTHomo sapiens 89Pro Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 Ile Gln Ser Leu Ile Ser Cys Trp Ala 20 25 9012PRTHomo sapiens 90Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu 1 5 10 9113PRTHomo sapiens 91Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 9214PRTHomo sapiens 92Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 9315PRTHomo sapiens 93Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15 9416PRTHomo sapiens 94Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 15 9517PRTHomo sapiens 95Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 15 Gln 9618PRTHomo sapiens 96Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 15 Gln Ser 9719PRTHomo sapiens 97Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 15 Gln Ser Leu 9820PRTHomo sapiens 98Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 15 Gln Ser Leu Ile 20 9921PRTHomo sapiens 99Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 15 Gln Ser Leu Ile Ser 20 10022PRTHomo sapiens 100Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 15 Gln Ser Leu Ile Ser Cys 20 10123PRTHomo sapiens 101Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 15 Gln Ser Leu Ile Ser Cys Trp 20 10224PRTHomo sapiens 102Leu Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 15 Gln Ser Leu Ile Ser Cys Trp Ala 20 10311PRTHomo sapiens 103Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu 1 5 10 10412PRTHomo sapiens 104Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 10513PRTHomo sapiens 105Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 10614PRTHomo sapiens 106Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 10715PRTHomo sapiens 107Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 15 10816PRTHomo sapiens 108Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 15 10917PRTHomo sapiens 109Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 15 Ser 11018PRTHomo sapiens 110Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 15 Ser Leu 11119PRTHomo sapiens 111Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 15 Ser Leu Ile 11220PRTHomo sapiens 112Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 15 Ser Leu Ile Ser 20 11321PRTHomo sapiens 113Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 15 Ser Leu Ile Ser Cys 20 11422PRTHomo sapiens 114Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 15 Ser Leu Ile Ser Cys Trp 20 11523PRTHomo sapiens 115Gln Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 15 Ser Leu Ile Ser Cys Trp Ala 20 11610PRTHomo sapiens 116Asn Ile Ser Pro Trp Ile Leu Ala Val Leu 1 5 10 11711PRTHomo sapiens 117Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 11812PRTHomo sapiens 118Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 11913PRTHomo sapiens 119Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 12014PRTHomo sapiens 120Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 12115PRTHomo sapiens 121Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 15 12216PRTHomo sapiens 122Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 12317PRTHomo sapiens 123Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu 12418PRTHomo sapiens 124Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu Ile 12519PRTHomo sapiens 125Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu Ile Ser 12620PRTHomo sapiens 126Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu Ile Ser Cys 20 12721PRTHomo sapiens 127Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu Ile Ser Cys Trp 20 12822PRTHomo sapiens 128Asn Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu Ile Ser Cys Trp Ala 20 12910PRTHomo sapiens 129Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu 1 5 10 13011PRTHomo sapiens 130Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 13112PRTHomo sapiens 131Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 13213PRTHomo sapiens 132Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 13314PRTHomo sapiens 133Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 13415PRTHomo sapiens 134Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 15 13516PRTHomo sapiens 135Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 13617PRTHomo sapiens 136Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu 13718PRTHomo sapiens 137Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu Ile 13819PRTHomo sapiens 138Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu Ile Ser 13920PRTHomo sapiens 139Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu Ile Ser Cys 20 14021PRTHomo sapiens 140Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu Ile Ser Cys Trp 20 14122PRTHomo sapiens 141Tyr Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 Leu Ile Ser Cys Trp Ala 20 1429PRTHomo sapiens 142Ile Ser Pro Trp Ile Leu Ala Val Leu 1 5 14310PRTHomo sapiens 143Ile Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 10 14411PRTHomo sapiens 144Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 14512PRTHomo sapiens 145Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 14613PRTHomo sapiens 146Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 14714PRTHomo sapiens 147Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 14815PRTHomo sapiens 148Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 15 14916PRTHomo sapiens 149Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 10 15 15017PRTHomo sapiens 150Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 10 15 Ile 15118PRTHomo sapiens 151Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 10 15 Ile Ser 15219PRTHomo sapiens 152Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 10 15 Ile Ser Cys 15320PRTHomo sapiens 153Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 10 15 Ile Ser Cys Trp 20 15421PRTHomo sapiens 154Ile Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 10 15 Ile Ser Cys Trp Ala 20 1558PRTHomo sapiens 155Ser Pro Trp Ile Leu Ala Val Leu 1 5 1569PRTHomo sapiens 156Ser Pro Trp Ile Leu Ala Val Leu Thr 1 5 15710PRTHomo sapiens 157Ser Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 10 15811PRTHomo sapiens 158Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 15912PRTHomo sapiens 159Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 16013PRTHomo sapiens 160Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 16114PRTHomo sapiens 161Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 16215PRTHomo sapiens 162Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 10 15 16316PRTHomo sapiens 163Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile 1 5 10 15 16417PRTHomo sapiens 164Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Ser Ser Leu Ile 1 5 10 15 Ser 16518PRTHomo sapiens 165Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Ser Ser Leu Ile 1 5 10 15 Ser Cys 16619PRTHomo sapiens 166Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Ser Ser Leu Ile 1 5 10 15 Ser Cys Trp 16720PRTHomo sapiens 167Ser Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Ser Ser Leu Ile 1 5 10 15 Ser Cys Trp Ala 20 1687PRTHomo sapiens 168Pro Trp Ile Leu Ala Val Leu 1 5 1698PRTHomo sapiens 169Pro Trp Ile Leu Ala Val Leu Thr 1 5 1709PRTHomo sapiens 170Pro Trp Ile Leu Ala Val Leu Thr Leu 1 5 17110PRTHomo sapiens 171Pro Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 10 17211PRTHomo sapiens 172Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 17312PRTHomo sapiens 173Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 17413PRTHomo sapiens 174Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 17514PRTHomo sapiens 175Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 10 17615PRTHomo sapiens 176Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile 1 5 10 15 17716PRTHomo sapiens 177Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser 1 5 10 15 17817PRTHomo sapiens 178Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser 1 5 10 15 Cys 17918PRTHomo sapiens 179Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser 1 5 10 15 Cys Trp 18019PRTHomo sapiens 180Pro Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser 1 5 10 15 Cys Trp Ala 1816PRTHomo sapiens 181Trp Ile Leu Ala Val Leu 1 5 1827PRTHomo sapiens 182Trp Ile Leu Ala Val Leu Thr 1 5 1838PRTHomo sapiens 183Trp Ile Leu Ala Val Leu Thr Leu 1 5 1849PRTHomo sapiens 184Trp Ile Leu Ala Val Leu Thr Leu Gln 1 5 18510PRTHomo sapiens 185Trp Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 10 18611PRTHomo sapiens 186Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 18712PRTHomo sapiens 187Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 18813PRTHomo sapiens 188Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 10 18914PRTHomo sapiens 189Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile 1 5 10 19015PRTHomo sapiens 190Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser 1 5 10 15 19116PRTHomo sapiens 191Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser Cys 1 5 10 15 19217PRTHomo sapiens 192Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser Cys 1 5 10 15 Trp 19318PRTHomo sapiens 193Trp Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser Cys 1 5 10

15 Trp Ala 1945PRTHomo sapiens 194Ile Leu Ala Val Leu 1 5 1956PRTHomo sapiens 195Ile Leu Ala Val Leu Thr 1 5 1967PRTHomo sapiens 196Ile Leu Ala Val Leu Thr Leu 1 5 1978PRTHomo sapiens 197Ile Leu Ala Val Leu Thr Leu Gln 1 5 1989PRTHomo sapiens 198Ile Leu Ala Val Leu Thr Leu Gln Ile 1 5 19910PRTHomo sapiens 199Ile Leu Ala Val Leu Thr Leu Gln Ile Gln 1 5 10 20011PRTHomo sapiens 200Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser 1 5 10 20112PRTHomo sapiens 201Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu 1 5 10 20213PRTHomo sapiens 202Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile 1 5 10 20314PRTHomo sapiens 203Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser 1 5 10 20415PRTHomo sapiens 204Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser Cys 1 5 10 15 20516PRTHomo sapiens 205Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser Cys Trp 1 5 10 15 20617PRTHomo sapiens 206Ile Leu Ala Val Leu Thr Leu Gln Ile Gln Ser Leu Ile Ser Cys Trp 1 5 10 15 Ala 20718PRTHomo sapiens 207Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val 20819PRTHomo sapiens 208Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val Cys 20920PRTHomo sapiens 209Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val Cys Leu 20 21021PRTHomo sapiens 210Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val Cys Leu Val 20 21122PRTHomo sapiens 211Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val Cys Leu Val Gly 20 21223PRTHomo sapiens 212Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val Cys Leu Val Gly Phe 20 21324PRTHomo sapiens 213Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val Cys Leu Val Gly Phe Met 20 21425PRTHomo sapiens 214Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val Cys Leu Val Gly Phe Met Leu 20 25 21526PRTHomo sapiens 215Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val Cys Leu Val Gly Phe Met Leu Tyr 20 25 21627PRTHomo sapiens 216Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg 20 25 21728PRTHomo sapiens 217Glu Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe 1 5 10 15 Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg Met 20 25 21817PRTHomo sapiens 218Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val 21918PRTHomo sapiens 219Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val Cys 22019PRTHomo sapiens 220Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val Cys Leu 22120PRTHomo sapiens 221Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val Cys Leu Val 20 22221PRTHomo sapiens 222Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val Cys Leu Val Gly 20 22322PRTHomo sapiens 223Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val Cys Leu Val Gly Phe 20 22423PRTHomo sapiens 224Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val Cys Leu Val Gly Phe Met 20 22524PRTHomo sapiens 225Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val Cys Leu Val Gly Phe Met Leu 20 22625PRTHomo sapiens 226Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val Cys Leu Val Gly Phe Met Leu Tyr 20 25 22726PRTHomo sapiens 227Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val Cys Leu Val Gly Phe Met Leu Tyr Arg 20 25 22827PRTHomo sapiens 228Val Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala 1 5 10 15 Val Cys Leu Val Gly Phe Met Leu Tyr Arg Met 20 25 22916PRTHomo sapiens 229Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 23017PRTHomo sapiens 230Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 Cys 23118PRTHomo sapiens 231Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 Cys Leu 23219PRTHomo sapiens 232Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 Cys Leu Val 23320PRTHomo sapiens 233Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 Cys Leu Val Gly 20 23421PRTHomo sapiens 234Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 Cys Leu Val Gly Phe 20 23522PRTHomo sapiens 235Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 Cys Leu Val Gly Phe Met 20 23623PRTHomo sapiens 236Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 Cys Leu Val Gly Phe Met Leu 20 23724PRTHomo sapiens 237Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 Cys Leu Val Gly Phe Met Leu Tyr 20 23825PRTHomo sapiens 238Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 Cys Leu Val Gly Phe Met Leu Tyr Arg 20 25 23926PRTHomo sapiens 239Leu Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 Cys Leu Val Gly Phe Met Leu Tyr Arg Met 20 25 24015PRTHomo sapiens 240Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 15 24116PRTHomo sapiens 241Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 24217PRTHomo sapiens 242Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 Leu 24318PRTHomo sapiens 243Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 Leu Val 24419PRTHomo sapiens 244Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 Leu Val Gly 24520PRTHomo sapiens 245Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 Leu Val Gly Phe 20 24621PRTHomo sapiens 246Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 Leu Val Gly Phe Met 20 24722PRTHomo sapiens 247Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 Leu Val Gly Phe Met Leu 20 24823PRTHomo sapiens 248Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 Leu Val Gly Phe Met Leu Tyr 20 24924PRTHomo sapiens 249Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 Leu Val Gly Phe Met Leu Tyr Arg 20 25025PRTHomo sapiens 250Gly Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 Leu Val Gly Phe Met Leu Tyr Arg Met 20 25 25114PRTHomo sapiens 251Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 25215PRTHomo sapiens 252Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 15 25316PRTHomo sapiens 253Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 15 25417PRTHomo sapiens 254Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 15 Val 25518PRTHomo sapiens 255Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 15 Val Gly 25619PRTHomo sapiens 256Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 15 Val Gly Phe 25720PRTHomo sapiens 257Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 15 Val Gly Phe Met 20 25821PRTHomo sapiens 258Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 15 Val Gly Phe Met Leu 20 25922PRTHomo sapiens 259Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 15 Val Gly Phe Met Leu Tyr 20 26023PRTHomo sapiens 260Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 15 Val Gly Phe Met Leu Tyr Arg 20 26124PRTHomo sapiens 261Gly Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 15 Val Gly Phe Met Leu Tyr Arg Met 20 26213PRTHomo sapiens 262Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 26314PRTHomo sapiens 263Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 26415PRTHomo sapiens 264Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 15 26516PRTHomo sapiens 265Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 15 26617PRTHomo sapiens 266Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 15 Gly 26718PRTHomo sapiens 267Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 15 Gly Phe 26819PRTHomo sapiens 268Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 15 Gly Phe Met 26920PRTHomo sapiens 269Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 15 Gly Phe Met Leu 20 27021PRTHomo sapiens 270Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 15 Gly Phe Met Leu Tyr 20 27122PRTHomo sapiens 271Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 15 Gly Phe Met Leu Tyr Arg 20 27223PRTHomo sapiens 272Val Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 15 Gly Phe Met Leu Tyr Arg Met 20 27312PRTHomo sapiens 273Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 27413PRTHomo sapiens 274Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 27514PRTHomo sapiens 275Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 27615PRTHomo sapiens 276Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 15 27716PRTHomo sapiens 277Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 15 27817PRTHomo sapiens 278Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 15 Phe 27918PRTHomo sapiens 279Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 15 Phe Met 28019PRTHomo sapiens 280Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 15 Phe Met Leu 28120PRTHomo sapiens 281Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 15 Phe Met Leu Tyr 20 28221PRTHomo sapiens 282Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 15 Phe Met Leu Tyr Arg 20 28322PRTHomo sapiens 283Ile Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 15 Phe Met Leu Tyr Arg Met 20 28411PRTHomo sapiens 284Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 28512PRTHomo sapiens 285Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 28613PRTHomo sapiens 286Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 28714PRTHomo sapiens 287Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 28815PRTHomo sapiens 288Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 15 28916PRTHomo sapiens 289Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 15 29017PRTHomo sapiens 290Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 15 Met 29118PRTHomo sapiens 291Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 15 Met Leu 29219PRTHomo sapiens 292Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 15 Met Leu Tyr 29320PRTHomo sapiens 293Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 15 Met Leu Tyr Arg 20 29421PRTHomo sapiens 294Ala Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 15 Met Leu Tyr Arg Met 20 29510PRTHomo sapiens 295Gly Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 10 29611PRTHomo sapiens 296Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 29712PRTHomo sapiens 297Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 29813PRTHomo sapiens 298Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 29914PRTHomo sapiens 299Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 30015PRTHomo sapiens 300Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 15 30116PRTHomo sapiens 301Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 15 30217PRTHomo sapiens 302Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 15 Leu 30318PRTHomo sapiens 303Gly Gly Leu Val Gly Leu Ile

Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 15 Leu Tyr 30419PRTHomo sapiens 304Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 15 Leu Tyr Arg 30520PRTHomo sapiens 305Gly Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 15 Leu Tyr Arg Met 20 3069PRTHomo sapiens 306Gly Leu Val Gly Leu Ile Phe Ala Val 1 5 30710PRTHomo sapiens 307Gly Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 10 30811PRTHomo sapiens 308Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 30912PRTHomo sapiens 309Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 31013PRTHomo sapiens 310Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 31114PRTHomo sapiens 311Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 31215PRTHomo sapiens 312Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 15 31316PRTHomo sapiens 313Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu 1 5 10 15 31417PRTHomo sapiens 314Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu 1 5 10 15 Tyr 31518PRTHomo sapiens 315Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu 1 5 10 15 Tyr Arg 31619PRTHomo sapiens 316Gly Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu 1 5 10 15 Tyr Arg Met 3178PRTHomo sapiens 317Leu Val Gly Leu Ile Phe Ala Val 1 5 3189PRTHomo sapiens 318Leu Val Gly Leu Ile Phe Ala Val Cys 1 5 31910PRTHomo sapiens 319Leu Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 10 32011PRTHomo sapiens 320Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 32112PRTHomo sapiens 321Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 32213PRTHomo sapiens 322Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 32314PRTHomo sapiens 323Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 32415PRTHomo sapiens 324Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu 1 5 10 15 32516PRTHomo sapiens 325Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr 1 5 10 15 32617PRTHomo sapiens 326Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr 1 5 10 15 Arg 32718PRTHomo sapiens 327Leu Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr 1 5 10 15 Arg Met 3287PRTHomo sapiens 328Val Gly Leu Ile Phe Ala Val 1 5 3298PRTHomo sapiens 329Val Gly Leu Ile Phe Ala Val Cys 1 5 3309PRTHomo sapiens 330Val Gly Leu Ile Phe Ala Val Cys Leu 1 5 33110PRTHomo sapiens 331Val Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 10 33211PRTHomo sapiens 332Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 33312PRTHomo sapiens 333Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 33413PRTHomo sapiens 334Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 33514PRTHomo sapiens 335Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu 1 5 10 33615PRTHomo sapiens 336Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr 1 5 10 15 33716PRTHomo sapiens 337Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg 1 5 10 15 33817PRTHomo sapiens 338Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg 1 5 10 15 Met 3396PRTHomo sapiens 339Gly Leu Ile Phe Ala Val 1 5 3407PRTHomo sapiens 340Gly Leu Ile Phe Ala Val Cys 1 5 3418PRTHomo sapiens 341Gly Leu Ile Phe Ala Val Cys Leu 1 5 3429PRTHomo sapiens 342Gly Leu Ile Phe Ala Val Cys Leu Val 1 5 34310PRTHomo sapiens 343Gly Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 10 34411PRTHomo sapiens 344Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 34512PRTHomo sapiens 345Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 34613PRTHomo sapiens 346Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu 1 5 10 34714PRTHomo sapiens 347Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr 1 5 10 34815PRTHomo sapiens 348Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg 1 5 10 15 34916PRTHomo sapiens 349Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg Met 1 5 10 15 3505PRTHomo sapiens 350Leu Ile Phe Ala Val 1 5 3516PRTHomo sapiens 351Leu Ile Phe Ala Val Cys 1 5 3527PRTHomo sapiens 352Leu Ile Phe Ala Val Cys Leu 1 5 3538PRTHomo sapiens 353Leu Ile Phe Ala Val Cys Leu Val 1 5 3549PRTHomo sapiens 354Leu Ile Phe Ala Val Cys Leu Val Gly 1 5 35510PRTHomo sapiens 355Leu Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 10 35611PRTHomo sapiens 356Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 35712PRTHomo sapiens 357Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu 1 5 10 35813PRTHomo sapiens 358Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr 1 5 10 35914PRTHomo sapiens 359Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg 1 5 10 36015PRTHomo sapiens 360Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg Met 1 5 10 15 3614PRTHomo sapiens 361Ile Phe Ala Val 1 3625PRTHomo sapiens 362Ile Phe Ala Val Cys 1 5 3636PRTHomo sapiens 363Ile Phe Ala Val Cys Leu 1 5 3647PRTHomo sapiens 364Ile Phe Ala Val Cys Leu Val 1 5 3658PRTHomo sapiens 365Ile Phe Ala Val Cys Leu Val Gly 1 5 3669PRTHomo sapiens 366Ile Phe Ala Val Cys Leu Val Gly Phe 1 5 36710PRTHomo sapiens 367Ile Phe Ala Val Cys Leu Val Gly Phe Met 1 5 10 36811PRTHomo sapiens 368Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu 1 5 10 36912PRTHomo sapiens 369Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr 1 5 10 37013PRTHomo sapiens 370Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg 1 5 10 37114PRTHomo sapiens 371Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg Met 1 5 10 37216PRTHomo sapiens 372Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 37317PRTHomo sapiens 373Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu 37418PRTHomo sapiens 374Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu Gly 37519PRTHomo sapiens 375Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu Gly Leu 37620PRTHomo sapiens 376Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu Gly Leu Phe 20 37721PRTHomo sapiens 377Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu Gly Leu Phe Leu 20 37822PRTHomo sapiens 378Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu Gly Leu Phe Leu Trp 20 37923PRTHomo sapiens 379Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu Gly Leu Phe Leu Trp Phe 20 38024PRTHomo sapiens 380Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu Gly Leu Phe Leu Trp Phe Leu 20 38125PRTHomo sapiens 381Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu Gly Leu Phe Leu Trp Phe Leu Lys 20 25 38226PRTHomo sapiens 382Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg 20 25 38327PRTHomo sapiens 383Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg Glu 20 25 38415PRTHomo sapiens 384Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 15 38516PRTHomo sapiens 385Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 38617PRTHomo sapiens 386Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 Gly 38718PRTHomo sapiens 387Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 Gly Leu 38819PRTHomo sapiens 388Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 Gly Leu Phe 38920PRTHomo sapiens 389Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 Gly Leu Phe Leu 20 39021PRTHomo sapiens 390Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 Gly Leu Phe Leu Trp 20 39122PRTHomo sapiens 391Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 Gly Leu Phe Leu Trp Phe 20 39223PRTHomo sapiens 392Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 Gly Leu Phe Leu Trp Phe Leu 20 39324PRTHomo sapiens 393Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 Gly Leu Phe Leu Trp Phe Leu Lys 20 39425PRTHomo sapiens 394Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 Gly Leu Phe Leu Trp Phe Leu Lys Arg 20 25 39526PRTHomo sapiens 395Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 Gly Leu Phe Leu Trp Phe Leu Lys Arg Glu 20 25 39614PRTHomo sapiens 396Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 39715PRTHomo sapiens 397Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 15 39816PRTHomo sapiens 398Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 39917PRTHomo sapiens 399Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 Leu 40018PRTHomo sapiens 400Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 Leu Phe 40119PRTHomo sapiens 401Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 Leu Phe Leu 40220PRTHomo sapiens 402Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 Leu Phe Leu Trp 20 40321PRTHomo sapiens 403Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 Leu Phe Leu Trp Phe 20 40422PRTHomo sapiens 404Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 Leu Phe Leu Trp Phe Leu 20 40523PRTHomo sapiens 405Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 Leu Phe Leu Trp Phe Leu Lys 20 40624PRTHomo sapiens 406Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 Leu Phe Leu Trp Phe Leu Lys Arg 20 40725PRTHomo sapiens 407Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 Leu Phe Leu Trp Phe Leu Lys Arg Glu 20 25 40813PRTHomo sapiens 408Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 40914PRTHomo sapiens 409Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 41015PRTHomo sapiens 410Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 15 41116PRTHomo sapiens 411Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 15 41217PRTHomo sapiens 412Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 15 Phe 41318PRTHomo sapiens 413Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 15 Phe Leu 41419PRTHomo sapiens 414Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 15 Phe Leu Trp 41520PRTHomo sapiens 415Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 15 Phe Leu Trp Phe 20 41621PRTHomo sapiens 416Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 15 Phe Leu Trp Phe Leu 20 41722PRTHomo sapiens 417Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 15 Phe Leu Trp Phe Leu Lys 20 41823PRTHomo sapiens 418Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 15 Phe Leu Trp Phe Leu Lys Arg 20 41924PRTHomo sapiens 419Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 15 Phe Leu Trp Phe Leu Lys Arg Glu 20 42012PRTHomo sapiens 420Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 42113PRTHomo sapiens 421Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 42214PRTHomo sapiens 422Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 42315PRTHomo sapiens 423Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 15 42416PRTHomo sapiens 424Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 15 42517PRTHomo sapiens 425Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 15 Leu 42618PRTHomo sapiens 426Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 15 Leu Trp 42719PRTHomo sapiens 427Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 15 Leu Trp Phe 42820PRTHomo sapiens 428Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 15 Leu Trp Phe Leu 20

42921PRTHomo sapiens 429Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 15 Leu Trp Phe Leu Lys 20 43022PRTHomo sapiens 430Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 15 Leu Trp Phe Leu Lys Arg 20 43123PRTHomo sapiens 431Leu Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 15 Leu Trp Phe Leu Lys Arg Glu 20 43211PRTHomo sapiens 432Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 43312PRTHomo sapiens 433Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 43413PRTHomo sapiens 434Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 43514PRTHomo sapiens 435Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 43615PRTHomo sapiens 436Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 15 43716PRTHomo sapiens 437Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 15 43817PRTHomo sapiens 438Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 15 Trp 43918PRTHomo sapiens 439Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 15 Trp Phe 44019PRTHomo sapiens 440Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 15 Trp Phe Leu 44120PRTHomo sapiens 441Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 15 Trp Phe Leu Lys 20 44221PRTHomo sapiens 442Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 15 Trp Phe Leu Lys Arg 20 44322PRTHomo sapiens 443Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 15 Trp Phe Leu Lys Arg Glu 20 44410PRTHomo sapiens 444Leu Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 10 44511PRTHomo sapiens 445Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 44612PRTHomo sapiens 446Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 44713PRTHomo sapiens 447Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 44814PRTHomo sapiens 448Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 44915PRTHomo sapiens 449Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 15 45016PRTHomo sapiens 450Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 15 45117PRTHomo sapiens 451Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 15 Phe 45218PRTHomo sapiens 452Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 15 Phe Leu 45319PRTHomo sapiens 453Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 15 Phe Leu Lys 45420PRTHomo sapiens 454Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 15 Phe Leu Lys Arg 20 45521PRTHomo sapiens 455Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 15 Phe Leu Lys Arg Glu 20 4569PRTHomo sapiens 456Val Pro Leu Leu Leu Ser Leu Phe Val 1 5 45710PRTHomo sapiens 457Val Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 10 45811PRTHomo sapiens 458Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 45912PRTHomo sapiens 459Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 46013PRTHomo sapiens 460Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 46114PRTHomo sapiens 461Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 46215PRTHomo sapiens 462Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 15 46316PRTHomo sapiens 463Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 15 46417PRTHomo sapiens 464Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 15 Leu 46518PRTHomo sapiens 465Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 15 Leu Lys 46619PRTHomo sapiens 466Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 15 Leu Lys Arg 46720PRTHomo sapiens 467Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 15 Leu Lys Arg Glu 20 4688PRTHomo sapiens 468Pro Leu Leu Leu Ser Leu Phe Val 1 5 4699PRTHomo sapiens 469Pro Leu Leu Leu Ser Leu Phe Val Leu 1 5 47010PRTHomo sapiens 470Pro Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 10 47111PRTHomo sapiens 471Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 47212PRTHomo sapiens 472Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 47313PRTHomo sapiens 473Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 47414PRTHomo sapiens 474Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 47515PRTHomo sapiens 475Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 15 47616PRTHomo sapiens 476Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu 1 5 10 15 47717PRTHomo sapiens 477Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu 1 5 10 15 Lys 47818PRTHomo sapiens 478Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu 1 5 10 15 Lys Arg 47919PRTHomo sapiens 479Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu 1 5 10 15 Lys Arg Glu 4807PRTHomo sapiens 480Leu Leu Leu Ser Leu Phe Val 1 5 4818PRTHomo sapiens 481Leu Leu Leu Ser Leu Phe Val Leu 1 5 4829PRTHomo sapiens 482Leu Leu Leu Ser Leu Phe Val Leu Gly 1 5 48310PRTHomo sapiens 483Leu Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 10 48411PRTHomo sapiens 484Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 48512PRTHomo sapiens 485Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 48613PRTHomo sapiens 486Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 48714PRTHomo sapiens 487Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 48815PRTHomo sapiens 488Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu 1 5 10 15 48916PRTHomo sapiens 489Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys 1 5 10 15 49017PRTHomo sapiens 490Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys 1 5 10 15 Arg 49118PRTHomo sapiens 491Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys 1 5 10 15 Arg Glu 4926PRTHomo sapiens 492Leu Leu Ser Leu Phe Val 1 5 4937PRTHomo sapiens 493Leu Leu Ser Leu Phe Val Leu 1 5 4948PRTHomo sapiens 494Leu Leu Ser Leu Phe Val Leu Gly 1 5 4959PRTHomo sapiens 495Leu Leu Ser Leu Phe Val Leu Gly Leu 1 5 49610PRTHomo sapiens 496Leu Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 10 49711PRTHomo sapiens 497Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 49812PRTHomo sapiens 498Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 49913PRTHomo sapiens 499Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 50014PRTHomo sapiens 500Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu 1 5 10 50115PRTHomo sapiens 501Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys 1 5 10 15 50216PRTHomo sapiens 502Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg 1 5 10 15 50317PRTHomo sapiens 503Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg 1 5 10 15 Glu 5045PRTHomo sapiens 504Leu Ser Leu Phe Val 1 5 5056PRTHomo sapiens 505Leu Ser Leu Phe Val Leu 1 5 5067PRTHomo sapiens 506Leu Ser Leu Phe Val Leu Gly 1 5 5078PRTHomo sapiens 507Leu Ser Leu Phe Val Leu Gly Leu 1 5 5089PRTHomo sapiens 508Leu Ser Leu Phe Val Leu Gly Leu Phe 1 5 50910PRTHomo sapiens 509Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 51011PRTHomo sapiens 510Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 51112PRTHomo sapiens 511Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 51213PRTHomo sapiens 512Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu 1 5 10 51314PRTHomo sapiens 513Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys 1 5 10 51415PRTHomo sapiens 514Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg 1 5 10 15 51516PRTHomo sapiens 515Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg Glu 1 5 10 15 5165PRTHomo sapiens 516Ser Leu Phe Val Leu 1 5 5176PRTHomo sapiens 517Ser Leu Phe Val Leu Gly 1 5 5187PRTHomo sapiens 518Ser Leu Phe Val Leu Gly Leu 1 5 5198PRTHomo sapiens 519Ser Leu Phe Val Leu Gly Leu Phe 1 5 5209PRTHomo sapiens 520Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 52110PRTHomo sapiens 521Ser Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 10 52211PRTHomo sapiens 522Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 52312PRTHomo sapiens 523Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu 1 5 10 52413PRTHomo sapiens 524Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys 1 5 10 52514PRTHomo sapiens 525Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg 1 5 10 52615PRTHomo sapiens 526Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg Glu 1 5 10 15 5275PRTHomo sapiens 527Leu Phe Val Leu Gly 1 5 5286PRTHomo sapiens 528Leu Phe Val Leu Gly Leu 1 5 5297PRTHomo sapiens 529Leu Phe Val Leu Gly Leu Phe 1 5 5308PRTHomo sapiens 530Leu Phe Val Leu Gly Leu Phe Leu 1 5 5319PRTHomo sapiens 531Leu Phe Val Leu Gly Leu Phe Leu Trp 1 5 53210PRTHomo sapiens 532Leu Phe Val Leu Gly Leu Phe Leu Trp Phe 1 5 10 53311PRTHomo sapiens 533Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu 1 5 10 53412PRTHomo sapiens 534Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys 1 5 10 53513PRTHomo sapiens 535Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg 1 5 10 53614PRTHomo sapiens 536Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg Glu 1 5 10 5379PRTHuman immunodeficiency virus 537Arg Tyr Leu Lys Asp Gln Gln Leu Leu 1 5 5389PRTHomo sapiens 538Arg Met Phe Pro Asn Ala Pro Tyr Leu 1 5 5399PRTHomo sapiens 539Tyr Met Phe Pro Asn Ala Pro Tyr Leu 1 5 54019PRTHomo sapiens 540Arg Ser Asp Glu Leu Val Arg His His Asn Met His Gln Arg Asn Met 1 5 10 15 Thr Lys Leu 54122PRTHomo sapiens 541Pro Gly Cys Asn Lys Arg Tyr Phe Lys Leu Ser His Leu Gln Met His 1 5 10 15 Ser Arg Lys His Thr Gly 20 54219PRTHomo sapiens 542Ser Gly Gln Ala Arg Met Phe Pro Asn Ala Pro Tyr Leu Pro Ser Cys 1 5 10 15 Leu Glu Ser 54319PRTHomo sapiens 543Ser Gly Gln Ala Tyr Met Phe Pro Asn Ala Pro Tyr Leu Pro Ser Cys 1 5 10 15 Leu Glu Ser 5446PRTArtificial SequenceDescription of Artificial Sequence Synthetic 6xHis tag 544His His His His His His 1 5

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