XBP1, CD138 and CS1 peptides, pharmaceutical compositions comprising said peptides, and methods of using said peptides and compositions

Abstract

The features of the present disclosure are immunogenic XBP1-derived peptides, CD138-derived peptides, and CS1-derived peptides (and their pharmaceutical compositions), etc. The peptides can be used in a variety of methods, such as methods for inducing an immune response, methods for generating antibodies, and methods for treating cancer (e.g., breast cancer, colon cancer, pancreatic cancer, blood cancers, e.g., leukemia, or plasma cell disorders such as multiple myeloma or Waldenstrom macroglobulinemia). The peptides (and pharmaceutical compositions comprising the peptides) can be used, for example, in methods for treating pre-cancerous conditions such as smoldering multiple myeloma. The peptides can also be incorporated into MHC molecule multimer compositions and used, for example, in methods for detecting T cells in a cell population.

Description

[0001] This application is a divisional application of the patent application with the filing date of November 5, 2013, application number "201380067770.1" (international application number PCT / US2013 / 068582), and invention title "XBP1, CD138 and CS1 Peptides, Pharmaceutical Compositions Comprising the Peptides and Methods of Using the Peptides and Compositions".

[0002] This application claims the priority of U.S. Serial No. 61 / 722,446 filed on November 5, 2012 and U.S. Serial No. 61 / 790,780 filed on March 15, 2013, the entire contents of which are incorporated herein by reference.

[0003] Statement Regarding Federally Sponsored Research or Development

[0004] The research described in this application was funded by grant numbers PO1-78378, PO1-155258, and P50-100707, all of which were obtained from the National Institutes of Health. Accordingly, the government has certain rights in this invention.

[0005] Sequence Listing

[0006] This application contains a sequence listing, which has been electronically submitted in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy, created on December 9, 2013, is named O2017-7001WO_SL.txt and is 117,979 bytes in size. Background of the Invention

[0008] Some types of vaccines have been developed for the prevention of infectious diseases, including attenuated microorganisms, recombinant protein vaccines, and DNA vaccines. Recently, research has been conducted on the development of vaccine immunotherapies for the treatment of cancer patients. Summary of the Invention

[0010] The present disclosure relates to immunogenic peptides that bind to MHC class I molecules such as HLA-A molecules. Peptides derived from X-Box binding protein 1 (XBP1), CD138, and CD2 subgroup 1 (CS1) have been found to be immunogenic and can be used, for example, to induce an immune response against various cancer cells. In some embodiments, the peptides have an increased affinity for HLA-A molecules, increased stability in the peptide-binding cleft of HLA-A, and, in the case of MHC molecules, the ability to induce the activation and proliferation of T cells (e.g., effector memory T cells and / or central memory T cells) when expressed on the surface of cells (e.g., cancer cells).

[0011] In addition, combinations of these peptides have been found to induce broad-spectrum immune responses against target antigens, and such broad-spectrum responses are capable of overcoming most of the therapeutic obstacles, including, for example, the heterogeneity of tumor-associated antigen expression, the 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., combinations in pharmaceutical compositions) can provide enhanced immune responses against various cancers.

[0012] It will be apparent from the following description that the peptides (and their pharmaceutical compositions) can be used in a variety of applications, such as methods for inducing immune responses, methods for activating T cells (e.g., including effector memory T cells and / or central memory T cells), methods for generating antibodies, and methods for treating, for example, cancers (e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, such as AML or CML), multiple myeloma, Waldenstrom's Macroglobulinemia, and precancerous conditions such as smoldering multiple myeloma.

[0013] In one aspect, the present disclosure features peptides, e.g., XBP1 peptides, CD138 peptides, and CS-1 peptides, which have an affinity for a variety of MHC molecules (e.g., HLA-A molecules such as HLA-A2 and HLA-A24), increased stability in the peptide-binding clefts of a variety of MHC molecules (e.g., HLA-A2 and HLA-A24), and the ability to induce the activation and proliferation of T cells (e.g., effector memory T cells and / or central memory T cells) when expressed on the surface of cells (e.g., cancer cells) in the case of MHC molecules such as HLA-A2 or HLA-A24.

[0014] It will be apparent from the following description that the peptides (and their pharmaceutical compositions) can be used in a variety of applications, such as methods for inducing immune responses, methods for activating T cells (e.g., including effector memory T cells and / or central memory T cells), methods for generating antibodies, and methods for treating, for example, cancers (e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, such as AML or CML), multiple myeloma, and precancerous conditions such as smoldering multiple myeloma.

[0015] In one aspect, the present disclosure features isolated peptides that comprise 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 peptides can bind to major histocompatibility complex (MHC) molecules such as MHC class I or class II molecules. 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 the 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 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 comprises the 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 having any one of SEQ ID NOs: 51 - 536 and having three, two, or one substitution. The substitution can be conservative or non - conservative. In one embodiment, the peptide is for treating a subject suffering from or at risk of suffering from cancer, such as the cancers 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 (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).

[0016] In one embodiment, the peptide can be used to treat a subject suffering from a pre-cancerous condition (e.g., smoldering multiple myeloma).

[0017] In one embodiment, the cancer is a cancer described herein. For example, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromphobic carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma and large cell carcinoma)), genitourinary cancer, such as ovarian cancer (including fallopian tube cancer, endometrial cancer and peritoneal cancer), cervical cancer, prostate cancer and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), nerve and neuroglial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer (e.g., nasopharyngeal cancer). In one embodiment, the cancer is breast cancer (e.g., invasive lobular carcinoma, invasive ductal carcinoma, mixed lobular and ductal carcinoma, intraductal cribriform, invasive ductal and lobular carcinoma or invasive carcinoma). In one embodiment, the cancer is colon adenocarcinoma (e.g., mucinous adenocarcinoma).

[0018] 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. In one embodiment, the peptide consists of an amino acid sequence of any one of SEQ ID NOs: 51 - 536 and having three, two or one substituted amino acids. In one embodiment, the peptide consists of the amino acid sequence of any one of SEQ ID NOs: 51 - 536.

[0019] In one embodiment, the peptide is an unspliced XBP1 peptide of group C (see, for example, Table 3), e.g., an unspliced XBP-1 peptide comprising the amino acid sequence of any one of SEQ ID NOs: 51-206, or an unspliced XBP1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to 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 the amino acid sequence of any one of SEQ ID NOs: 51-536 or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to any one of SEQ ID NOs: 51-206. In one embodiment, the unspliced 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 any one of SEQ ID NOs: 51-206 and has an amino acid sequence with three, two or one substituted amino acids. The substitution can be conservative or non-conservative. In one embodiment, the unspliced XBP1 peptide of group C consists of the amino acid sequence of any one of SEQ ID NOs: 51-206.

[0020] In one embodiment, the peptide is a CD138 peptide of Group C, e.g., a CD138 peptide comprising the 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 to the amino acid sequence of 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 the 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 to 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 the amino acid sequence of any one of SEQ ID NOs: 207 - 371 and has an amino acid sequence with three, two, one substitution. The substitution can be conservative or non - conservative. In one embodiment, the CD138 peptide of Group C consists of the amino acid sequence of any one of SEQ ID NOs: 207 - 371.

[0021] In one embodiment, the peptide is a Group C CS-1 peptide, e.g., a CS-1 peptide comprising the 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 to the 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 the 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 to any one of SEQ ID NOs: 372-536. In one embodiment, the Group C CS-1 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 any one of SEQ ID NOs: 372-536 and has an amino acid sequence with three, two, one substitution. The substitution can be conservative or non-conservative. In one embodiment, the Group C CS-1 peptide consists of the amino acid sequence of any one of SEQ ID NOs: 372-536.

[0022] In one embodiment, the peptides of group C, e.g., XBP peptides, CD138 peptides, and / or CS-1 peptides are used to treat a subject suffering from cancer or at risk of developing cancer, such as the cancers described herein, e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML. In one embodiment, the peptides of group C are used to treat a subject suffering from a pre-cancerous condition, e.g., smoldering multiple myeloma. In one embodiment, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma, and large cell carcinoma)), genitourinary cancer, e.g., ovarian cancer (including fallopian tube cancer, endometrial cancer, and peritoneal cancer), cervical cancer, prostate cancer, and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer, or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), nerve and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer (e.g., nasopharyngeal cancer).

[0023] In some embodiments, any of the isolated peptides described herein can bind to major histocompatibility complex (MHC) molecules (e.g., MHC class I or class II molecules). The MHC molecules can be, for example, human MHC molecules. The MHC molecules can be, for example, HLA-A molecules, HLA-B molecules, and / or HLA-C molecules. Preferably, the MHC molecule is one or more HLA-A molecules (e.g., HLA-A1, HLA-A2, HLA-A3, and HLA-A24).

[0024] In one aspect, the present disclosure features peptides derived from immunogenic X-Box binding protein 1 (XBP1), peptides derived from CD138, and peptides derived from CD2 subgroup 1 (CS1), e.g., which have increased affinity for HLA-A2 molecules, increased stability in the peptide-binding cleft of HLA-A2, and, in the case of MHC molecules, the ability to induce activation and proliferation of T cells (e.g., effector memory T cells and / or central memory T cells) when expressed on the surface of cells (e.g., cancer cells). For example, in the case of MHC molecules, all or a subgroup of these peptides are expressed on the surface of a variety of cancer cells, including multiple myeloma cells, and particularly smoldering multiple myeloma cells, colon cancer cells, breast cancer cells, pancreatic cancer cells, prostate cancer cells, and white blood cells, such as acute myeloid leukemia (AML) cells, and the presence of these peptides induces activation and proliferation of T cells against these and other cancers.

[0025] In addition, it has been found that combinations of these peptides can induce a broad-spectrum immune response against target antigens, and that this broad-spectrum response is capable of overcoming most therapeutic obstacles, including, for example, heterogeneity in tumor-associated antigen expression, frequent mutation of specific antigens, and variability in the human T cell repertoire in an individual. Thus, administration of combinations of these peptides (e.g., in a pharmaceutical composition) can provide an enhanced immune response against various cancers.

[0026] It will be apparent from the following description that the peptides (and their pharmaceutical compositions) can be used in a variety of applications, such as methods for inducing an immune response, methods for activating T cells (e.g., including effector memory T cells and / or central memory T cells), methods for generating antibodies, and methods for treating, for example, cancers (e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, such as AML or CML), multiple myeloma, and pre-cancerous conditions such as smoldering multiple myeloma.

[0027] 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 the 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 the 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 the 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 the amino acid sequence of any one of SEQ ID NOs: 1-18 and has an amino acid sequence with three, two, or one substitution. The substitution can be conservative or non-conservative. In one embodiment, the peptide can be used to treat a subject suffering from or at risk of suffering from cancer, such as the cancers described herein, such as breast cancer (e.g., invasive lobular carcinoma, invasive ductal carcinoma, mixed lobular and ductal carcinoma, intraductal cribriform carcinoma, invasive ductal and lobular carcinoma, or invasive carcinoma), colon cancer (e.g., adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma.

[0028] In one embodiment, the peptide is used to treat a subject suffering from a pre-cancerous condition, such as, smoldering multiple myeloma. In one embodiment, the cancer is a cancer described herein. For example, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma and large cell carcinoma)), genitourinary cancer, such as ovarian cancer (including fallopian tube cancer, endometrial cancer and peritoneal cancer), cervical cancer, prostate cancer and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), nerve and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer (e.g., nasopharyngeal cancer).

[0029] 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 one of the amino acid sequences of SEQ ID NOs: 1-18 and has three, two or one substitution. In one embodiment, the peptide consists of the amino acid sequence of any one of SEQ ID NOs: 1-18.

[0030] In one embodiment, the peptide is an unspliced XBP1 peptide of group A (see, e.g., Table 1), e.g., an unspliced XBP-1 peptide comprising the amino acid sequence of any one of SEQ ID NOs: 1-6, an unspliced XBP1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence of any one of SEQ ID NOs: 1-6. In one embodiment, the peptide has a length of 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids and comprises the 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 to the amino acid sequence of any one of SEQ ID NOs: 1-6. In one embodiment, the unspliced XBP1 peptide of group A has a length of 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids, and the peptide comprises the amino acid sequence of any one of SEQ ID NOs: 1-6 and has an amino acid sequence with three, two or one substituted amino acids. The substitution can be conservative or non-conservative. In one embodiment, the unspliced XBP1 peptide of group A consists of the amino acid sequence of any one of SEQ ID NOs: 1-6.

[0031] In one embodiment, the peptide is a spliced XBP1 peptide of Group A, for example, a spliced XBP-1 peptide comprising the 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 to the amino acid sequence of any one of SEQ ID NOs: 7-10. In one embodiment, the peptide has a length of 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids and comprises the 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 to the amino acid sequence of any one of SEQ ID NOs: 7-10. In one embodiment, the spliced XBP1 peptide of Group A has a length of 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids, and the peptide comprises the amino acid sequence of any one of SEQ ID NOs: 7-10 and has a three, two, one substituted amino acid sequence. The substitution can be conservative or non-conservative. In one embodiment, the spliced XBP1 peptide of Group A consists of the amino acid sequence of any one of SEQ ID NOs: 7-10.

[0032] In one embodiment, the peptide is a CD138 peptide of Group A, for example, a CD138 peptide comprising the 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 to the amino acid sequence of any one of SEQ ID NOs: 11-14. In one embodiment, the length of the peptide is 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids, and comprises the 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 to the amino acid sequence of any one of SEQ ID NOs: 11-14. In one embodiment, the CD138 peptide of Group A has a length of 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids, and the peptide comprises an amino acid sequence having the amino acid sequence of any one of SEQ ID NOs: 11-14 and having three, two, one substituted amino acid sequence. The substitution may be conservative or non-conservative. In one embodiment, the CD138 peptide of Group A consists of the amino acid sequence of any one of SEQ ID NOs: 11-14.

[0033] In one embodiment, the peptide is a CS-1 peptide of Group A, for example, a CS-1 peptide comprising the 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 to the 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 the 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 to the 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 having the amino acid sequence of any one of SEQ ID NOs: 15-18 and having three, two, one substituted amino acid sequence. The substitution may be conservative or non-conservative. In one embodiment, the CS-1 peptide of Group A consists of the amino acid sequence of any one of SEQ ID NOs: 15-18.

[0034] In one embodiment, the peptides of Group A, e.g., unspliced XBP1 peptide, spliced XBP1 peptide, CD138 peptide, and / or CS-1 peptide, are used to treat a subject suffering from cancer or at risk of developing cancer, such as the cancers described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML. In one embodiment, the peptides of Group A, e.g., unspliced XBP1 peptide, spliced XBP1 peptide, CD138 peptide, and / or CS-1 peptide, are used to treat a subject suffering from a pre-cancerous condition, such as smoldering multiple myeloma. In one embodiment, the cancer is a cancer described herein. For example, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma, and large cell carcinoma)), genitourinary cancer, e.g., ovarian cancer (including fallopian tube cancer, endometrial cancer, and peritoneal cancer), cervical cancer, prostate cancer, and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer, or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), nerve and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer.

[0035] In some embodiments, any of the isolated peptides described herein can bind to a major histocompatibility complex (MHC) molecule (e.g., MHC class I or class II molecule). The MHC molecule can be, e.g., an HLA-A2 molecule. The MHC molecule can be, for example, a human MHC molecule.

[0036] In another aspect, the present disclosure features peptides derived from immunogenic X-Box protein 1 (XBP1), peptides derived from CD138, and peptides derived from CD2 subgroup 1 (CS1) that, for example, have increased affinity for HLA-A24 molecules, increased stability in the peptide-binding groove of HLA-A24, and, in the case of MHC molecules, the ability to induce activation and proliferation of T cells (e.g., effector memory T cells and / or central memory T cells) when expressed on the surface of cells (e.g., cancer cells). For example, all or a subgroup of these peptides are expressed on the surface of a variety of cancer cells, including multiple myeloma cells, colon cancer cells, breast cancer cells, pancreatic cancer cells, prostate cancer cells, and white blood cells, such as acute myeloid leukemia (AML) cells, and the presence of these peptides induces activation and proliferation of T cells against these and other cancers.

[0037] It will be apparent from the following description that the peptides (and their pharmaceutical compositions) can be used in a variety of applications, such as methods for inducing an immune response, methods for activating T cells (e.g., including effector memory T cells and / or central memory T cells), methods for generating antibodies, and methods for treating, for example, cancer (e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, such as AML or CML), multiple myeloma, and pre-cancerous conditions such as smoldering multiple myeloma.

[0038] In one aspect, the disclosure features an isolated peptide that comprises 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 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 the amino acid sequence of any 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 the 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 the amino acid sequence of any 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 the amino acid sequence of any of SEQ ID NOs: 29-50 and has an amino acid sequence with three, two, or one substitution. The substitution can be conservative or non-conservative. In one embodiment, the peptide is for treating a subject having cancer or at risk of having cancer, e.g., the cancers described herein, e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer (e.g., invasive lobular carcinoma, invasive ductal carcinoma, mixed lobular and ductal carcinoma, intraductal cribriform carcinoma, invasive ductal and lobular carcinoma, or invasive carcinoma), colon cancer (e.g., colonic adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma.

[0039] In one embodiment, the peptide is used to treat a subject suffering from a pre-cancerous condition, such as, smoldering multiple myeloma. In one embodiment, the cancer is a cancer described herein. For example, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma and large cell carcinoma)), genitourinary cancer, such as ovarian cancer (including fallopian tube cancer, endometrial cancer and peritoneal cancer), cervical cancer, prostate cancer and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer (e.g., nasopharyngeal cancer).

[0040] 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 and having three, two or one substituted amino acids. In one embodiment, the peptide consists of the amino acid sequence of any one of SEQ ID NOs: 29-50.

[0041] In one embodiment, the peptide is an unspliced XBP1 peptide from Group B (see Table 2), e.g., an unspliced XBP-1 peptide comprising the amino acid sequence of any one of SEQ ID NOs: 29 and 33-37, or an unspliced XBP1 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to 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 the 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 to any one of SEQ ID NOs: 29 and 33-37. In one embodiment, the unspliced 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 any one of SEQ ID NOs: 29 and 33-37 and has an amino acid sequence with three, two, one substituted amino acids. The substitution can be conservative or non-conservative. In one embodiment, the unspliced XBP1 peptide is from the 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 at the C-terminus of SEQ ID NO: 29 in SEQ ID NO: 19. In one embodiment, the unspliced XBP1 peptide is from the 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 at the N-terminus of SEQ ID NO: 33 in SEQ ID NO: 19, and / or 1, 2, 3 or more (e.g., 1) amino acids at the C-terminus of SEQ ID NO: 33 in SEQ ID NO: 19.In one embodiment, the unspliced XBP1 peptide is from the amino acid sequence of 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, 4) amino acids at the N-terminus of 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 at the C-terminus of SEQ ID NO: 36 in SEQ ID NO: 19. In one embodiment, the unspliced XBP1 peptide is from the amino acid sequence SEQ ID NO: 19 and comprises the amino acid sequence of any one of SEQ ID NO: 34, 35 or 37 and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3) amino acids at the N-terminus and / or C-terminus of any one of SEQ ID NO: 34, 35 or 37 in SEQ ID NO: 19. In one embodiment, the unspliced XBP1 peptide from Group B consists of the amino acid sequence of any one of SEQ ID NO: 29 and 33 - 37.

[0042] In one embodiment, the peptide is a spliced XBP1 peptide from Group B, e.g., a spliced XBP-1 peptide comprising the amino acid sequence of any one of SEQ ID NO: 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 to any one of SEQ ID NO: 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 the amino acid sequence of any one of SEQ ID NO: 30, 38, and 39, or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to any one of SEQ ID NO: 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 any one of SEQ ID NO: 30, 38, and 39 and has an amino acid sequence with three, two, or one substituted amino acid. The substitution can be conservative or non-conservative. In one embodiment, the spliced XBP1 peptide is from the amino acid sequence of SEQ ID NO: 20 and comprises the amino acid sequence of any one of SEQ ID NO: 30, 38, and 39, and 1, 2, 3, 4, 5, 6, 7, 8, 9, or more (e.g., 1, 2, 3) amino acids at the N-terminus and / or C-terminus of any one of SEQ ID NO: 30, 38, and 39 in SEQ ID NO: 20. In one embodiment, the spliced XBP1 peptide from Group B consists of the amino acid sequence of any one of SEQ ID NO: 30, 38, and 39.

[0043] In one embodiment, the peptide is a CD138 peptide from Group B, e.g., a CD138 peptide comprising the amino acid sequence of any one of SEQ ID NO:31, or a CD138 peptide comprising an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to any one of SEQ ID NO:31 and 40 - 45. In one embodiment, the peptide has a length of 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids and comprises the amino acid sequence of any one of SEQ ID NO:31 and 40 - 45, or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to any one of SEQ ID NO:31 and 40 - 45. In one embodiment, the CD138 peptide from Group B has a length of 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids, and the peptide comprises any one of SEQ ID NO:31 and 40 - 45 and has an amino acid sequence with three, two, one substituted amino acids. The substitution can be conservative or non - conservative. In one embodiment, the CD138 peptide is from the 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 at the N - terminus of SEQ ID NO:31 in SEQ ID NO:21. In one embodiment, the CD138 peptide is from the 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 at the N - terminus of 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 at the C - terminus of SEQ ID NO:42 or 44 in SEQ ID NO:21.In one embodiment, the CD138 peptide is derived from the 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 at the N-terminus of SEQ ID NO: 45 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 or 6) amino acids at the C-terminus of SEQ ID NO: 45 in SEQ ID NO: 21. In one embodiment, the CD138 peptide is derived from the amino acid sequence SEQ ID NO: 21, and comprises the amino acid sequence of any one of SEQ ID NO: 40, 41 or 43, and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3) amino acids at the N-terminus and / or C-terminus of any one of SEQ ID NO: 40, 41 or 43 in SEQ ID NO: 21. In one embodiment, the CD138 peptide from Group B consists of the amino acid sequence of SEQ ID NO: 31 and any one of SEQ ID NO: 40 - 45.

[0044] In one embodiment, the peptide is a CS-1 peptide from Group B, e.g., a CS-1 peptide comprising the amino acid sequence of any one of SEQ ID NO: 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 to any one of SEQ ID NO: 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 the amino acid sequence of any one of SEQ ID NO: 32 and 46-50, or an amino acid sequence having 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to any one of SEQ ID NO: 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 having any one of SEQ ID NO: 32 and 46-50 and having three, two, one substituted amino acids. The substitution can be conservative or non-conservative. In one embodiment, the CS-1 peptide is derived from the amino acid sequence SEQ ID NO: 22 and comprises the amino acid sequence of SEQ ID NO: 32 and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3) amino acids at the C-terminus of SEQ ID NO: 32 in SEQ ID NO: 22. In one embodiment, the CS-1 peptide is derived from the amino acid sequence SEQ ID NO: 22 and comprises the amino acid sequence of any one of SEQ ID NO: 46-50 and 1, 2, 3, 4, 5, 6, 7, 8, 9 or more (e.g., 1, 2, 3) amino acids at the N-terminus and / or C-terminus of any one of SEQ ID NO: 46-50 in SEQ ID NO: 22. In one embodiment, the CS-1 peptide from Group B consists of the amino acid sequence of any one of SEQ ID NO: 32 and 46-50.

[0045] In one embodiment, the peptides of Group B, e.g., unspliced XBP1 peptide, spliced XBP1 peptide, CD138 peptide, and / or CS-1 peptide, are used to treat a subject suffering from cancer or at risk of developing cancer, e.g., the cancers described herein, e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML. In one embodiment, the peptides of Group B, e.g., unspliced XBP1 peptide, spliced XBP1 peptide, CD138 peptide, and / or CS-1 peptide, are used to treat a subject suffering from a pre-cancerous condition, e.g., smoldering multiple myeloma. In one embodiment, the cancer is the cancer described herein. For example, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma, and large cell carcinoma)), genitourinary cancer, e.g., ovarian cancer (including fallopian tube cancer, endometrial cancer, and peritoneal cancer), cervical cancer, prostate cancer, and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer, or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), nerve and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer (e.g., nasopharyngeal cancer).

[0046] In some embodiments, any of the isolated peptides described herein can be associated with a major histocompatibility complex (MHC) molecule that is recognized by an antigen-specific T cell receptor on a T cell.

[0047] In another aspect, the present disclosure features a fusion protein comprising a first amino acid sequence consisting of a peptide described herein (e.g., an unspliced XBP1 peptide from Group A, B, or C, a spliced XBP1 peptide from Group A or B, a CD138 peptide from Group A, B, or C, and / or a CS-1 peptide from Group A, B, or C, such as those described herein); and a second amino acid sequence that is heterologous to the first amino acid sequence.

[0048] In some embodiments, the second amino acid sequence can comprise, or can be, a targeting polypeptide, an immunostimulatory molecule, an immunoglobulin or an antigen-binding fragment thereof, an Fc receptor-binding region of an immunoglobulin molecule, or a carrier polypeptide. The targeting polypeptide can be, for example, a polypeptide that targets an isolated peptide to an antigen-presenting cell (e.g., a dendritic cell, macrophage, monocyte, or B cell). The immunostimulatory molecule can be, for example, a cytokine or a T helper epitope. The immunoglobulin can be, for example, a single-chain Fv immunoglobulin fragment or an entire immunoglobulin molecule. The carrier polypeptide can comprise, or can be, a KLH (keyhole limpet hemocyanin) polypeptide, or an albumin polypeptide.

[0049] In some embodiments, any isolated peptide described herein can contain a linker sequence. The linker sequence can directly or indirectly link the first amino acid sequence to the second amino acid sequence. The linker sequence can comprise or consist of one or more amino acids, for example, 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 ten or more) protease cleavage sites.

[0050] In some embodiments, the second amino acid sequence can be at the amino terminus or carboxyl terminus of the first amino acid sequence.

[0051] In some embodiments, any isolated peptide or fusion protein described herein can be detectably labeled. The detectable label can be selected from the group consisting of a luminescent label, a fluorescent label, a radioactive label, and an enzyme label.

[0052] In yet another aspect, the present disclosure features: (i) an isolated nucleic acid encoding any isolated peptide 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 a eukaryotic cell. The cultured cell can be, for example, 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 immune cell described herein.

[0053] In another aspect, the present disclosure features a method of producing a peptide. The method involves the step of culturing any cultured cell described herein under conditions that permit expression of the peptide. The method can further comprise the step of isolating the peptide from the cell or from the medium in which the cell is cultured.

[0054] In another aspect, the present disclosure features a pharmaceutical composition comprising any one or more 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.

[0055] In one embodiment, the composition comprises at least two peptides. For example, the composition comprises an unspliced XBP1 peptide, e.g., an unspliced XBP1 peptide from Group A, and a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group A; the composition comprises an unspliced XBP1 peptide, e.g., an unspliced XBP1 peptide from Group A, and a CD138 peptide, e.g., a CD138 peptide from Group A; the composition comprises an unspliced XBP1 peptide, e.g., an unspliced 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.

[0056] In one embodiment, the composition comprises at least three peptides. For example, the composition comprises an unspliced XBP1 peptide (e.g., the unspliced XBP1 peptide described from Group A), a spliced XBP1 peptide (e.g., the spliced XBP1 peptide from Group A), and a CD138 peptide (e.g., the CD138 peptide from Group A); the composition comprises an unspliced XBP1 peptide (e.g., the unspliced XBP1 peptide described from Group A), a spliced XBP1 peptide (e.g., the spliced XBP1 peptide from Group A), and a CS-1 peptide (e.g., the CS-1 peptide from Group A); the composition comprises an unspliced XBP1 peptide (e.g., the unspliced XBP1 peptide described from Group A), a CD138 peptide (e.g., the CD138 peptide from Group A), and a CS-1 peptide (e.g., the CS-1 peptide from Group A); the composition comprises a spliced XBP1 peptide (e.g., the spliced XBP1 peptide from Group A), a CD138 peptide (e.g., the CD138 peptide from Group A), and a CS-1 peptide (e.g., the CS-1 peptide from Group A). In one embodiment, the composition comprises at least three peptides, e.g., an unspliced XBP1 peptide (e.g., the unspliced XBP1 peptide from Group A), a spliced XBP1 peptide (e.g., the spliced XBP-1 peptide from Group A), and a CD138 peptide (e.g., the CD138 peptide from Group A).

[0057] In one embodiment, the composition comprises four peptides. For example, the composition comprises an unspliced XBP1 peptide (e.g., the unspliced XBP1 peptide described from Group A), a spliced XBP1 peptide (e.g., the spliced XBP1 peptide from Group A), a CD138 peptide (e.g., the CD138 peptide from Group A), and a CS-1 peptide (e.g., the CS-1 peptide from Group A).

[0058] In one embodiment, the composition comprises unspliced XBP1 peptides from group A, which are 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprise the amino acid sequence of any one of SEQ ID NO: 1-6, for example, SEQ ID NO: 6. In one embodiment, the composition comprises spliced XBP1 peptides from group A, which are 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprise the amino acid sequence of any one of SEQ ID NO: 7-10, for example, SEQ ID NO: 10. In one embodiment, the composition comprises CD138 peptides from group A, which are 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprise the amino acid sequence of any one of SEQ ID NO: 11-14, for example, SEQ ID NO: 12. In one embodiment, the composition comprises CS-1 peptides from group A, which are 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprise the amino acid sequence of any one of SEQ IDNO: 15-18, for example, SEQ ID NO: 16.

[0059] In one embodiment, the composition comprises four peptides, and the four peptides are peptides from group A that comprise (for example, are composed of) the amino acid sequence of SEQ IDNO: 6, peptides from group A that comprise (for example, are composed of) the amino acid sequence of SEQ ID NO: 10, peptides from group A that comprise (for example, are composed of) the amino acid sequence of SEQ ID NO: 12, and peptides from group A that comprise (for example, are composed of) the amino acid sequence of SEQ ID NO: 16.

[0060] In one embodiment, the composition comprises at least two peptides. For example, the composition comprises an unspliced XBP1 peptide, e.g., an unspliced XBP1 peptide from Group B, and a spliced XBP1 peptide, e.g., a spliced XBP1 peptide from Group B; the composition comprises an unspliced XBP1 peptide, e.g., an unspliced XBP1 peptide from Group B, and a CD138 peptide, e.g., a CD138 peptide from Group B; the composition comprises an unspliced XBP1 peptide, e.g., an unspliced 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.

[0061] In one embodiment, the composition comprises at least three peptides. For example, the composition comprises an unspliced XBP1 peptide, e.g., the described unspliced XBP1 peptide 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 an unspliced XBP1 peptide, e.g., an unspliced 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 an unspliced XBP1 peptide, e.g., an unspliced 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., an unspliced XBP1 peptide (e.g., an unspliced 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).

[0062] In one embodiment, the composition comprises four peptides. For example, the composition comprises an unspliced XBP1 peptide, e.g., an unspliced 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.

[0063] In one embodiment, the composition comprises unspliced XBP1 peptides from group B, which are 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprise the amino acid sequence of any one of SEQ ID NO: 29 and 33 - 37. In one embodiment, the composition comprises spliced XBP1 peptides from group B, which are 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprise the amino acid sequence of any one of SEQ ID NO: 30, 38 and 39. In one embodiment, the composition comprises CD138 peptides from group B, which are 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprise the amino acid sequence of any one of SEQ ID NO: 31 and 40 - 45. In one embodiment, the composition comprises CS-1 peptides from group B, which are 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, up to 25, 30 or 35 amino acids in length and comprise the amino acid sequence of any one of SEQ ID NO: 32 and 46 - 50.

[0064] In one embodiment, the composition comprises four peptides, and the four peptides are peptides from group B that comprise (such as consisting of) the amino acid sequence of any one of SEQ ID NO: 29 and 33 - 37, peptides from group B that comprise (such as consisting of) the amino acid sequence of any one of SEQ ID NO: 30, 38 and 39, peptides from group B that comprise (such as consisting of) the amino acid sequence of any one of SEQ ID NO: 31 and 40 - 45, and peptides from group B that comprise (such as consisting of) the amino acid sequence of any one of SEQ ID NO: 32 and 46 - 50.

[0065] 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.

[0066] The composition may also include, for example, one or more additional agents, such as one or more therapeutic, diagnostic, or prophylactic agents, or immunostimulatory or immunomodulatory agents. Immunostimulatory agents include, but are not limited to, for example, T helper epitopes, altered peptide ligands, adjuvants, or any other immunostimulatory agent described herein. T helper epitopes can be, for example, the PADRE sequence or the universal tetanus toxoid T helper cell (TT Th) epitope. Adjuvants can be selected from the group consisting of: Freund's complete adjuvant, Freund's incomplete adjuvant, alum, ligands of Toll receptors, saponins (e.g., QS21), RIBI, cholera toxin (CT), Escherichia coli heat-labile toxin (LT), mutant CT (MCT), mutant Escherichia coli heat-labile toxin (MLT), adjuvants comprising carboxymethylcellulose, polyinosinic:polycytidylic acid and poly-L-lysine double-stranded RNA (e.g., poly IC-LC, e.g., hiltonol), adjuvants comprising water and oil emulsions (e.g., montanide), and adjuvants comprising proteins (e.g., cytokines, complement, GCSF, GM-CSF). In one embodiment, the immunostimulatory 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., cytokines, complement, GCSF, GM-CSF. In one embodiment, the immunomodulatory agent can be a protein, e.g., an antibody that modulates the immune system. For example, an antibody that modulates the immune system can be an anti-CTLA4 antibody, e.g., ipilimumab or tremelimumab, an anti-PD-1 antibody, or an anti-PDL-1 antibody. In one embodiment, the immunomodulatory agent can be a small molecule adjuvant, e.g., thalidomide or a thalidomide derivative, e.g., lenalidomide.

[0067] The composition may further include immunogenic peptides other than the peptides disclosed above, for example, immunogenic peptides from WT1 or derivatives thereof. Exemplary WT1 peptides are described in U.S. Patent No. 7,598,221, the content of which is incorporated herein by reference. In one embodiment, the composition comprises one or more immunogenic peptides from WT1 or derivatives thereof, for example, one or more selected from the following: WT1 class I epitopes; peptides comprising or consisting of RMFPNAPYL (SEQ ID NO:538) (WT1 126-134); peptides comprising or consisting of YMFPNAPYL (SEQ ID NO:539); peptides comprising or consisting of RSDELVRHHNMHQRNMTKL (SEQ ID NO:540) (WT1 427-445); peptides comprising or consisting of PGCNKRYFKLSHLQMHSRKHTG (SEQ ID NO:541) (WT1 331-352); peptides comprising or consisting of SGQARMFPNAPYLPSCLES (SEQ ID NO:542) (WT1 122-140); and peptides comprising or consisting of SGQAYMFPNAPYLPSCLES (SEQ ID NO:543). Other immunogenic peptides include, but are not limited to, immunogenic peptides from MUC1, immunogenic peptides from gp100, immunogenic peptides from TRP-2, immunogenic peptides from MAG1, immunogenic peptides from NY-ESO1, immunogenic peptides from HER-2; and immunogenic peptides from AIM2.

[0068] In one embodiment, the compositions described herein are used to treat a subject suffering from cancer or at risk of developing cancer, such as the cancers described herein, such as breast cancer (e.g., invasive lobular carcinoma, invasive ductal carcinoma, mixed lobular and ductal carcinoma, intraductal cribriform carcinoma, invasive ductal and lobular carcinoma, invasive carcinoma), colon cancer (e.g., colonic adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML or multiple myeloma. In one embodiment, the compositions described herein are used to treat a subject suffering from a pre-cancerous condition, such as smoldering multiple myeloma. In one embodiment, the cancer is a cancer described herein. For example, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma and large cell carcinoma)), genitourinary cancer, such as ovarian cancer (including fallopian tube cancer, endometrial cancer and peritoneal cancer), cervical cancer, prostate cancer and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer.

[0069] In another aspect, the present disclosure features a kit comprising: (i) any one or more isolated peptides from Group A, from Group B, and / or from Group C; and instructions for administering the peptide to a subject, such as a subject suffering from cancer (e.g., cancers described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, multiple myeloma), or a subject suffering from a pre-cancerous condition (e.g., smoldering multiple myeloma); (ii) a composition described herein and instructions for administering the peptide to a subject, such as a subject suffering from cancer (e.g., cancers described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, multiple myeloma), or a subject suffering from a pre-cancerous condition (e.g., smoldering multiple myeloma); and / or (iii) one or more isolated nucleic acids encoding an isolated peptide, one or more vectors comprising the isolated nucleic acid, or one or more cultured cells comprising the vector, and instructions for producing the isolated peptide. In one embodiment, the cancer is a cancer described herein. For example, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma, and large cell carcinoma)), genitourinary cancer, such as ovarian cancer (including fallopian tube cancer, endometrial cancer, and peritoneal cancer), cervical cancer, prostate cancer, and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer, or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), nerve and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer.

[0070] In some embodiments, the kit may further include, for example, one or more pharmaceutically acceptable carriers, one or more immune stimulants or modulators, or one or more therapeutic, diagnostic, or prophylactic agents. In one embodiment, the immune stimulant is the immune stimulant described herein. The one or more immune stimulants may be selected from the group consisting of: T helper epitopes, altered peptide ligands, and adjuvants. In one embodiment, the immune stimulant is an adjuvant comprising carboxymethyl cellulose, 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, complement, GCSF, GM-CSF). In one embodiment, the immune modulator is the immune modulator described herein, e.g., a protein, e.g., an antibody that 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 includes instructions for co-administering the immune stimulant and / or immune modulator with the peptide or the composition described herein.

[0071] In one embodiment, the kit further contains additional immunogenic peptides, e.g., an immunogenic peptide from Wt1 or a derivative thereof, e.g., the 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 MAG1, an immunogenic peptide from NY-ESO1, an immunogenic peptide from HER-2, an immunogenic peptide from AIM2. In one embodiment, the kit further contains instructions for co-administering the additional immunogenic peptide (e.g., the WT1 peptide) with the peptide or the composition described herein.

[0072] In another aspect, the present disclosure features an article of manufacture comprising: a container, and a composition contained within the container, wherein the composition is the composition described herein. The container may have a label indicating that the composition is for inducing an immune response in a mammal (e.g., a human). The label may further indicate that the composition is to be administered to a mammal suffering from cancer, suspected of having cancer, or at risk of developing cancer, the cancer being, for example, the cancer described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma, or to a patient suffering from a pre-cancerous condition, e.g., smoldering multiple myeloma. The article of manufacture may further include instructions for administering the composition to a mammal (e.g., a human). The composition may be, for example, in solution, dry, or lyophilized.

[0073] In one embodiment, the cancer is a cancer described herein. For example, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma and large cell carcinoma)), genitourinary cancer, such as ovarian cancer (including fallopian tube cancer, endometrial cancer and peritoneal cancer), cervical cancer, prostate cancer and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), nerve and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer.

[0074] In yet another aspect, the present disclosure features methods for inducing an immune response in a subject, the methods comprising the step of delivering (e.g., administering) to the subject one or more of any of the isolated peptides described herein and / or the compositions described herein. In one embodiment, at least two, e.g., 2, 3, or 4 peptides from Group A are administered to the subject. For example, one or more of the unspliced XBP1 peptide from Group A, the spliced XBP1 peptide from Group A, the CD138 peptide from Group A, the CS-1 peptide from Group A, and combinations thereof may be administered to the subject. In one embodiment, the unspliced XBP1 peptide from Group A (e.g., the spliced XBP1 peptide comprising SEQ ID NO:6), the spliced XBP1 peptide from Group A (e.g., the spliced XBP1 peptide comprising SEQ ID NO:10), the CD138 peptide from Group A (e.g., the CD138 peptide comprising SEQ ID NO:12), and the CS-1 peptide from Group A (e.g., the CS-1 peptide comprising SEQ ID NO:16) are administered to the subject. In one embodiment, at least two, e.g., 2, 3, or 4 peptides from Group B are administered to the subject. For example, one or more of the unspliced XBP1 peptide from Group B, the spliced XBP1 peptide from Group B, the CD138 peptide from Group B, the CS-1 peptide from Group B, and combinations thereof may be administered to the subject. In one embodiment, the unspliced XBP1 peptide from Group B (e.g., the unspliced XBP1 peptide comprising SEQ ID NO:29), the spliced XBP1 peptide from Group B (e.g., the CD138 peptide comprising SEQ ID NO:30), the CD138 peptide from Group B (e.g., the CD138 peptide comprising SEQ ID NO:31), and the CS-1 peptide from Group B (e.g., the CS-1 peptide comprising SEQ ID NO:32) are administered to the subject. In one embodiment, 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) peptides from Group C are administered to the subject. In another embodiment, two or more peptides from Group A and Group C or Group B and Group C are administered to the subject.

[0075] The method may further include determining whether an immune response has occurred in a subject after delivering one or more peptides or compositions to the subject. The one or more peptides may be delivered to the subject as a pharmaceutical composition, e.g., a pharmaceutical composition described herein. The subject may be, e.g., a mammal (e.g., a human) or any other subject described herein. The subject may have cancer, be suspected of having cancer, or be at risk of developing cancer, such as 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 pre-cancerous condition, e.g., smoldering multiple myeloma. In one embodiment, the cancer is a cancer described herein. For example, the cancer may be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma, and large cell carcinoma)), genitourinary cancer, e.g., ovarian cancer (including fallopian tube cancer, endometrial cancer, and peritoneal cancer), cervical cancer, prostate cancer, and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer, or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer.

[0076] In some embodiments, the method may include determining whether one or more cancer cells express one or more of XBP1, CD138, or CS-1.

[0077] In some embodiments, the method may further comprise administering to the subject one or more additional therapies, e.g., chemotherapeutic agents, ionizing radiation, surgery, or one or more additional immunotherapeutic agents. One or more forms of ionizing radiation can be, for example, γ-radiation, X-radiation, or β-radiation. One or more chemotherapeutic agents can be the chemotherapeutic agents described herein, e.g., chemotherapeutic agents selected from the group consisting of platinum-based agents, taxanes, topoisomerase inhibitors, antimetabolites, alkylating agents, protease inhibitors, and vinca alkaloids. Exemplary chemotherapeutic agents include, but are not limited to: cisplatin, carboplatin, procarbazine, mechlorethamine, cyclophosphamide, camptothecin, adriamycin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, mitomycin, etoposide, verampil, podophyllotoxin, taxol, transplatinum, 5-fluorouracil, vincristine, vinblastine, methotrexate, and analogs of any of the foregoing. The method may further comprise administering to the subject one or more immunostimulatory agents, e.g., one or more immunostimulatory agents described herein.

[0078] In one embodiment, the method further comprises administering an additional immunogenic peptide together with one or more peptides described herein, e.g., an immunogenic peptide from WT1 or a derivative thereof, e.g., an immunogenic WT1 as described herein. Other immunogens 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 MAG1, an immunogenic peptide from NY-ESO1, an immunogenic peptide from HER-2, an immunogenic peptide from AIM2.

[0079] In some embodiments, delivery comprises administering to a subject one or more peptides from Group A, Group B, and / or Group C, or a composition described herein. In some embodiments, delivery comprises administering to a subject one or more nucleic acids, each comprising a nucleotide sequence encoding one or more peptides, the nucleotide sequence operably linked to an expression control sequence. The nucleic acid can be in a recombinant cell that has been transfected with and expresses the one or more peptides. The recombinant cell can be a transfected cell prepared by transfecting a cell obtained from the subject, or a progeny of the transfected cell. The recombinant cell can be an antigen-presenting cell, such as, but not limited to, a dendritic cell, macrophage, monocyte, or B cell.

[0080] In some embodiments of any of the methods described above, delivery comprises: contacting one or more peptides with a cell; and, after contacting the one or more peptides with the cell, delivering the cell to the subject. The cell can be, for example, an antigen-presenting cell, such as any of those described herein. The cell can be, for example, a cell obtained from the subject, or a progeny of the cell. In some embodiments, the cell can be a cell obtained from another subject of the same species as the subject, or a progeny of the cell. The other subject can express at least one MHC molecule that is the same as that of the subject. The at least one MHC molecule can be, for example, an MHC class I molecule such as an HLA-A2 molecule and / or an HLA-A24 molecule.

[0081] In another aspect, the present disclosure features methods for treating a subject having cancer (e.g., cancers described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma) or a pre-cancerous condition, e.g., a subject having smoldering multiple myeloma. In one embodiment, the method comprises administering to the subject any 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 the peptides from Group A or a composition described herein, wherein the subject has cancer or is at risk of developing cancer, the cancer being, e.g., cancers described herein, e.g., breast cancer (e.g., invasive lobular carcinoma, invasive ductal carcinoma, mixed lobular and ductal carcinoma, intraductal cribriform carcinoma, invasive lobular and ductal carcinoma, invasive carcinoma), colon cancer (e.g., colonic adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML or multiple myeloma, or the subject has a pre-cancerous condition, e.g., smoldering multiple myeloma. In one embodiment, the method comprises administering to the subject any one or more (e.g., one, two, three, or four) of the peptides from Group B, or a composition described herein, wherein the subject has cancer or is at risk of developing cancer, the cancer being the cancers described herein, e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer (e.g., invasive lobular carcinoma, invasive ductal carcinoma, mixed lobular and ductal carcinoma, intraductal cribriform carcinoma, invasive lobular and ductal carcinoma, invasive carcinoma), colon cancer (e.g., colonic adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma. In one embodiment, the method comprises administering to the subject any 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 the peptides from Group C or a composition described herein, wherein the subject has cancer or is at risk of developing cancer, e.g., cancers described herein, e.g., breast cancer (e.g., invasive lobular carcinoma, invasive ductal carcinoma, mixed lobular and ductal carcinoma, intraductal cribriform carcinoma, invasive lobular and ductal carcinoma, invasive carcinoma), colon cancer (e.g., colonic adenocarcinoma, e.g., mucinous adenocarcinoma), pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, or multiple myeloma, or the subject has a pre-cancerous condition, e.g., smoldering multiple myeloma.

[0082] In one embodiment, the cancer is the cancer described herein. For example, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell cancer (e.g., papillary renal cell cancer, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma, and large cell carcinoma)), genitourinary cancer, such as ovarian cancer (including fallopian tube cancer, endometrial cancer, and peritoneal cancer), cervical cancer, prostate cancer, and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer, or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer.

[0083] In one embodiment, at least two, three, or four peptides from Group A are administered to the subject. For example, two or more of the unspliced XBP1 peptide from Group A, the spliced XBP1 peptide from Group A, the CD138 peptide from Group A, and the CS-1 peptide from Group A can be administered to the subject, and combinations thereof. In one embodiment, an unspliced XBP1 peptide from Group A (e.g., the unspliced XBP1 peptide comprising SEQ ID NO:6), a spliced XBP1 peptide from Group A (e.g., the spliced XBP1 peptide comprising SEQ ID NO:10), a CD138 peptide from Group A (the CD138 peptide comprising SEQ ID NO:12), and a CS-1 peptide (the CS-1 peptide comprising SEQ ID NO:16) are administered to the subject. One or more peptides can be delivered to the subject as a pharmaceutical composition (e.g., a pharmaceutical composition from Group A).

[0084] In one embodiment, at least two, e.g., 2, 3, or 4, peptides from Group B are administered to a subject. For example, an unspliced 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 can be administered to a subject. In one embodiment, an unspliced XBP1 peptide from Group B (e.g., an unspliced 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 (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) are administered to a subject.

[0085] In one embodiment, 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) peptides from Group C are administered to a subject. In another embodiment, two or more peptides from Group A and Group C or Group B and Group C are administered to a subject.

[0086] In one embodiment, the method further comprises administering to the subject an additional agent, e.g., a chemotherapeutic agent and / or an immune stimulant and / or an immunomodulator. In one embodiment, the additional agent is an immune stimulant, e.g., an immune stimulant as described herein. In one embodiment, the immune stimulant 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), and an adjuvant comprising a protein (e.g., a cytokine, GCSF, GM-CSF). In one embodiment, the additional agent is an immunomodulator, e.g., an immunomodulator as described herein. In one embodiment, the immunomodulator is a protein, e.g., an antibody that 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 together with one or more peptides, e.g., an immunogenic peptide from WT1 or a derivative thereof, e.g., a WT1 peptide 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 MAG1, an immunogenic peptide from NY-ESO1, an immunogenic peptide from HER-2; and an immunogenic peptide from AIM2.

[0087] In one embodiment, the method further comprises one or more additional doses of a peptide from Group A, Group B, and / or Group C or a composition as described herein. In one embodiment, one or more additional doses are administered to the subject about 14 days after a previous administration, e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 doses of a peptide from Group A, Group B, and / or Group C or a composition as described herein are administered to the subject every other week.

[0088] In another aspect, the present disclosure features a method for selecting a treatment for a mammal in need thereof. The method includes the steps of determining whether one or more cancer cells of cancer in the mammal express XBP1, e.g., cancers described herein, e.g., breast cancer cells, colon cancer cells, pancreatic cancer cells, prostate cancer cells, blood cells, e.g., plasma cells; and if one or more cancer cells express XBP1, selecting one or more peptides from Group A, Group B, and / or Group C, a fusion protein comprising the peptide, or a composition described herein as a therapeutic agent for the mammal. The method further includes the step of delivering to the subject one or more peptides from Group A, Group B, and / or Group C, a fusion protein comprising the peptide, or a composition described herein after determining that one or more cells of the cancer express XBP1.

[0089] In another aspect, the present disclosure features a method for selecting a treatment for a mammal suffering from cancer. The method includes the steps of determining whether one or more cancer cells of the cancer in the mammal express CD138, e.g., cancers described herein, e.g., breast cancer cells, colon cancer cells, pancreatic cancer cells, prostate cancer cells, blood cells, e.g., plasma cells; and if one or more cancer cells express CD138, selecting one or more peptides from Group A, Group B, and / or Group C, a fusion peptide comprising the peptide, or a composition described herein as a therapeutic agent for the mammal. The method may further include the step of delivering to the subject one or more peptides from Group A, Group B, and / or Group C, a fusion protein comprising the peptide, or a composition described herein after determining that one or more cells of the cancer express CD138.

[0090] In another aspect, the present disclosure features a method for selecting a treatment for a mammal in need thereof. The method includes determining whether one or more cancer cells of the cancer in the mammal express CS-1, e.g., cancers described herein, e.g., breast cancer cells, colon cancer cells, pancreatic cancer cells, prostate cancer cells, blood cells, e.g., plasma cells; and if one or more of the cancer cells express CS-1, selecting one or more peptides from Group A, Group B, and / or Group C, a fusion protein comprising the peptide, or a composition described herein as a therapeutic agent for the mammal. The composition may further include the step of delivering to the subject one or more peptides from Group A, Group B, and / or Group C, a fusion protein comprising the peptide, or a composition described herein after determining that one or more cells of the cancer express CS1.

[0091] In another aspect, the present disclosure features a method for selecting a therapeutic agent for a mammal suffering from cancer, such as the cancers described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer, or a pre-cancerous condition, e.g., smoldering multiple myeloma. The method includes the steps of selecting one or more peptides from Group A, Group B, and / or Group C, a fusion protein comprising the peptide, or a composition described herein as a therapeutic agent for the mammal if one or more cancer cells of the mammal express XBP1, CD138, and / or CS-1. The method may further include the step of delivering to the subject one or more peptides from Group A, Group B, and / or Group C, a fusion protein comprising the peptide, or a composition described herein after determining that one or more cells of the cancer express XBP1, CD138, and / or CS-1.

[0092] In some embodiments of any of the above methods, the subject or mammal may be one that has been treated for cancer (such as the cancers described herein, e.g., breast cancer, colon cancer, pancreatic cancer, prostate cancer) and is non-responsive to the treatment, e.g., the peptides from Group A, Group B, and / or Group C, the fusion protein comprising the peptide, or the composition described herein may be a second-line, third-line, or fourth-line treatment.

[0093] In another aspect, the present disclosure features the compositions 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 ten or more) peptide-binding regions of the MHC molecule. In some embodiments, each peptide-binding region has a peptide from Group A, Group B, and / or Group C bound thereto. In some embodiments, each peptide-binding region has a peptide from Group A, Group B, and / or Group C non-covalently or covalently bound thereto. The MHC molecule multimer may comprise two or more (e.g., two, three, four, five, six, seven, eight, nine, or 10 or more) complete MHC molecules. The MHC molecule multimer may comprise human MHC molecules. The MHC molecule multimer may comprise MHC class I molecules such as HLA-A molecules, e.g., HLA-A2 molecules or HLA-A24 molecules.

[0094] In some embodiments, two or more peptide-binding regions can be from the same MHC molecule. In some embodiments, two or more peptide-binding regions are from different MHC molecules. In some embodiments, two or more peptide-binding regions can be 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) region from a different MHC molecule.

[0095] In some embodiments, the MHC molecule multimer is capable of binding to at least one of one or more peptides of the composition.

[0096] In some embodiments, the composition can be detectably labeled. For example, one or more peptides and / or one or more peptide-binding regions can be detectably labeled. In some embodiments, at least one of one or more MHC molecule multimers or at least one of one or more peptides is detectably labeled.

[0097] 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 belongs. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, but 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.

[0098] Other features and advantages of the present invention, e.g., methods for inducing an immune response in a subject, will become apparent from the following specification, drawings, and claims. Brief Description of the Drawings

[0100] Figure 1a is a bar graph showing the HLA-A2 binding ability 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 58-66 (IVMP 58-66 ; GILGFVFTL) (SEQ ID NO:25) was used as an HLA-A2 specific positive control peptide.

[0101] Figure 1b is a bar graph showing the HLA-A2 stability of a multipeptide 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 protein58-66 (IVMP 58-66 ; GILGFVFTL) (SEQ ID NO:25) is used as an HLA-A2 specific positive control peptide.

[0102] Figure 2 are a series of bar graphs showing different phenotypes of multi-peptide specific CTLs (MP-CTLs). The Y-axis represents the percentage of cells in a given population.

[0103] Figure 3 are a series of bar graphs showing MP-CTL responses to HLA-A2 + MM cell lines in terms of IFN-γ production. The Y-axis represents the percentage of IFN-γ+ cells in a given population.

[0104] Figure 4 shows the induction of MP-CTL proliferation by stimulation with HLA-A2+ MM cells (including primary myeloma cell lines and multiple myeloma cell lines). The upper panel is a representative dot plot from flow cytometry analysis. The Y-axis represents CD8 expression, and the X-axis represents the reduction in CFSE staining, which is a direct measure of cell proliferation. The lower panel is a bar graph showing the MP-CTL proliferative responses to primary multiple myeloma cells (lower left panel) and multiple myeloma cell lines (lower right panel). The Y-axis in the bar graph represents the percentage of proliferating MP-CTLs, and the X-axis represents the source of the stimulatory MM cells tested.

[0105] Figure 5 are a series of graphs showing the cytotoxic activity of MP-CTLs against HLA-A2+ MM cells (including primary MM cells and cell lines). The Y-axis represents percentage cytotoxicity, and the X-axis represents the ratio of effector cells (MP-CTLs) to target cells.

[0106] Figure 6a are a series of dot plots showing the peptide-specific responses of multi-peptide specific CTLs generated from a single donor (donor A). The Y-axis represents the CD107α expression level, and the X-axis represents the IFN-γ expression level.

[0107] Figure 6b are a series of bar graphs showing the peptide-specific responses of multi-peptide specific CTLs generated from three donors (donor B, donor C, and donor D). The Y-axis represents the percentage of CD107α+ cells (upper panel) or the percentage of IFN-γ+ cells (lower panel), and the X-axis represents the peptides presented by K562-A2 cells.

[0108] Figure 7 is a table showing the relative expression of unspliced XBP1 and spliced XBP1 in multiple cancer cell lines. The relative expression levels are represented using plus or minus signs and also numbers indicating the number of plus signs.

[0109] Figure 8a A series of bar graphs showing the proliferative responses of XBP1-CTL against HLA-A2+ breast cancer cell lines on day 6. The X-axis represents the reduction in CFSE staining, which is a direct measure of cell proliferation.

[0110] Figure 8b A series of bar graphs showing the proliferative responses of XBP1-CTL against HLA-A2+ breast cancer cell lines on day 7. The X-axis represents the reduction in CFSE staining, which is a direct measure of cell proliferation.

[0111] Figure 9 A series of dot plots showing IFN-γ production and cell activation (CD69 expression) of XBP1-CTL against HLA-A2+ breast cancer cell lines. The Y-axis represents CD69 expression and the X-axis represents IFN-γ expression.

[0112] Figure 10 A series of dot plots showing degranulation (CD107α) of XBP1-CTL against HLA-A2+ breast cancer cell lines. The Y-axis represents CD107α expression and the X-axis represents CD8 expression.

[0113] Figure 11 A series of graphs showing the proliferative responses of XBP1-CTL against HLA-A2+ pancreatic cancer cell lines and colon cancer cell lines. The X-axis represents the reduction in CFSE staining, which is a direct measure of cell proliferation.

[0114] Figure 12 A series of graphs showing IFN-γ production and degranulation responses of XBP1-CTL against HLA-A2+ pancreatic cancer cell lines and colon cancer cell lines. The Y-axis represents CD107α expression and the X-axis represents IFN-γ expression.

[0115] Figure 13 A table showing relative CD138 expression in multiple cancer cell lines. The relative expression levels are represented using plus or minus signs and also numbers indicating the number of plus signs.

[0116] Figure 14 A table showing relative CS1 expression in multiple cancer cell lines. The relative expression levels are represented using plus or minus signs and also numbers indicating the number of plus signs.

[0117] Figure 15aa and b show the increase in CD8+ CTLs induced by a mixture of immunogenic XBP1-unspliced, XBP1-spliced, CD138, and CS-1 HLA-A2-specific peptides from T cells of different patients with smoldering multiple myeloma. The Y-axis in the left panel represents the percentage of CD3+CD8+ CTLs, and the Y-axis in the right panel represents the percentage of CD4+ Th cells. The X-axis represents the number of peptide stimulations before phenotypic analysis.

[0118] Figure 16a are a series of bar graphs showing the proliferative responses of MP-CTLs generated from patients with smoldering multiple myeloma to myeloma cells in an HLA-A2-restricted manner. The X-axis represents the decrease in CFSE staining as a direct measure of cell proliferation. The responses 5 days after stimulation are shown in the upper panel, the responses 6 days after are shown in the middle panel, and the responses 7 days after are shown in the lower panel.

[0119] Figure 16b are a series of bar graphs showing the proliferative responses of MP-CTLs generated from a second patient with smoldering multiple myeloma to myeloma cells in an HLA-A2-restricted manner. The Y-axis represents the number of cells, and the X-axis represents CFSE staining. The responses 5 days after stimulation are shown in the upper panel, the responses 6 days after are shown in the middle panel, and the responses 7 days after are shown in the lower panel.

[0120] Figure 17a are a series of dot plots showing IFN-γ production by MP-CTLs generated from patients with smoldering multiple myeloma in response to myeloma cell lines in an HLA-A2-restricted manner. The Y-axis represents IFN-γ expression, and the X-axis represents CD8 expression.

[0121] Figure 17b are a series of bar graphs showing IFN-γ production by MP-CTLs generated from four patients with smoldering multiple myeloma in response to myeloma cell lines in an HLA-A2-restricted manner. The Y-axis represents the percentage of IFN-γ+ cells, and the X-axis represents the type of cells used to stimulate the MP-CTLs.

[0122] Figure 18a are a series of dot plots showing degranulation by MP-CTLs generated from patients with smoldering multiple myeloma in response to myeloma cell lines in an HLA-A2-restricted manner. The Y-axis represents CD107α expression, and the X-axis represents CD8 expression.

[0123] Figure 18b are a series of bar graphs showing degranulation by MP-CTLs generated from patients with smoldering multiple myeloma in response to myeloma cell lines in an HLA-A2-restricted manner. The Y-axis represents the percentage of CD107α cells, and the X-axis represents the type of cells used to stimulate the MP-CTLs.

[0124] Figure 19a A series of dot plots showing the multifunctional IFN-γ production and degranulation (CD107α) of K562-A2 cells responsive to presentation of each peptide in CD3+CD8+CD137+ MP-CTLs generated from patients with smoldering multiple myeloma. The Y-axis represents CD107α expression and the Y-axis represents + cells and the X-axis represents IFN-γ expression.

[0125] Figure 19b A series of bar graphs showing the multifunctional IFN-γ production and degranulation (CD107α) of K562-A2 cells responsive to presentation of each peptide in CD3+CD8+CD137+ MP-CTLs generated from three patients with smoldering multiple myeloma. The Y-axis represents the percentage of CD3+CD8+CD137+ that express both IFN-γ and CD107α, and the X-axis represents the peptides presented by K562 A2 + cells.

[0126] Figure 19c A summary bar graph showing the multifunctional IFN-γ production and degranulation (CD107α) of K562-A2 cells responsive to presentation of each peptide in CD3+CD8+CD137+ MP-CTLs generated from three patients with smoldering multiple myeloma. The Y-axis represents the percentage of CD3+CD8+CD137+ that express both IFN-γ and CD107α, and the X-axis represents the peptides presented by K562 A2 + cells.

[0127] Figure 19d A summary bar graph of IFN-γ production and degranulation (CD107α) both from two different experiments of K562-A2 cells responsive to presentation of each peptide in CD3+CD8+CD137+ MP-CTLs generated from three patients with smoldering multiple myeloma. The Y-axis represents the percentage of CD3+CD8+CD137+ cells that express both IFN-γ and CD107α, and the X-axis represents the peptides presented by K562 A2 + cells.

[0128] Figure 20a A series of dot plots (upper panel) and bar graphs (lower panel) showing the increase in memory CD8 + T cells in MP-CTLs generated from four patients with smoldering multiple myeloma. The Y-axis in the dot plot shows CCR7 expression and the X-axis represents CD45RP expression. The Y-axis in the bar graph represents the percentage of cells positive for the T cell subsets defined on the X-axis.

[0129] Figure 20bA series of bar graphs showing the increase in effector memory (EM) cells in MP-CTLs generated from two patients with smoldering multiple myeloma. The increase in the number of peptide stimulations from 4 to 7 led to an increase in the percentage of EM-type cells. The Y-axis in the bar graphs represents the percentage of cells positive for the T cell subsets defined on the X-axis.

[0130] Figure 20c A series of bar graphs showing the increase in IFN-γ CD107α+ in effector memory cells (EM) and terminal effector cells (TE) from three patients with smoldering multiple myeloma. The Y-axis represents the percentage of double-positive IFN-γ+ CD107α+ cells, and the X-axis represents the cell types used to stimulate the MP-CTLs.

[0131] Figure 20d A series of bar graphs showing the CD69 activation of untreated and memory CD8+ T cell subsets in MP-CTLs generated from four patients with smoldering multiple myeloma in response to MM cell lines (HLA-A2+ U266 cells or HLA-A2 RPMI cells). The Y-axis shows the percentage of CD69+ cells, and the X-axis represents the CTL subsets.

[0132] Figure 21 Shows the affinity of peptides from unspliced XBP1, spliced XBP1, CD138, and CS1 for HLA-A24. T2 cells were exposed to the indicated peptides at a concentration of 1 mg / ml. HIV envelope protein 583-591 (RYLKDQQLL; SEQ ID NO:537) was used as an HLA-A24-specific positive control peptide.

[0133] Figure 22 Shows 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 the peptides at the indicated concentrations.

[0134] Figure 23 Shows 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 the peptides at the indicated concentrations.

[0135] Figure 24 Shows the affinity of CS1 peptides 3 (SEQ ID NO:48) and 5 (SEQ ID NO:32) for HLA-A24. T2 cells were exposed to the peptides at the indicated concentrations.

[0136] Figure 25It is a diagram of a method for generating peptide-specific CTLs. APCs presenting a specified peptide are used to stimulate CD3+ T lymphocytes from a donor to generate peptide-specific CTLs.

[0137] Figure 26a It 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-axis represents the percentage of CD8+ T lymphocytes, and the X-axis represents the number of peptide stimulations before phenotypic analysis.

[0138] Figure 26b It 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-axis represents the percentage of CD8+ T lymphocytes, and the X-axis represents the number of peptide stimulations before phenotypic analysis.

[0139] Figure 26c It 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-axis represents the percentage of CD8+ T lymphocytes, and the X-axis represents the number of peptide stimulations before phenotypic analysis.

[0140] Figure 26d It 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-axis represents the percentage of CD8+ T lymphocytes, and the X-axis represents the number of peptide stimulations before phenotypic analysis.

[0141] Figure 27 It is a schematic diagram of a method for evaluating the response of peptide-specific CTLs to various multiple myeloma tumor cells. The peptide-specific CTLs are incubated with KMS, OPM1, or U266 multiple myeloma cells for 5 hours, and IFN-γ production, CD107α upregulation, CD8 T cell proliferation, or IL-2 production is measured.

[0142] Figure 28 It is a series of dot plots showing IFN-γ production and CD8 expression of peptide-specific CTLs in response to multiple myeloma cells. The Y-axis represents the level of IFN-γ expression and the X-axis indicates CD8 expression. The peptide specificity of the CTL population analyzed is shown in the left panel. The CD8+ population and IFN-γ+ population are shown in the boxed area.

[0143] Figure 29a It is a series of dot plots showing IFN-γ expression in populations and subpopulations of CTLs. CTLs specific for unspliced XBP1 peptide 7 (SEQ ID NO:29) were stimulated with KMS11 cells. The populations presented in the boxed area are shown in the figure.

[0144] Figure 29b A series of dot plots showing IFN-γ expression in populations and subpopulations of CTLs. CTLs specific for spliced XBP1 peptide 1 (SEQ ID NO:30) were stimulated with KMS11 cells. The populations presented in the boxed regions are shown in the figure.

[0145] Figure 29c A series of dot plots showing IFN-γ expression in populations and subpopulations of CTLs. CTLs specific for CD138 peptide 1 (SEQ ID NO:31) were stimulated with KMS11 cells. The populations presented in the boxed regions are shown in the figure.

[0146] Figure 29d A series of dot plots showing IFN-γ expression in populations and subpopulations of CTLs. CTLs specific for CS1 peptide 5 (SEQ ID NO:32) were stimulated with KMS11 cells. The populations presented in the boxed regions are shown in the figure.

[0147] Figure 30 Schematic diagram of a CD107α detection assay for measuring degranulation. Release of lytic granules due to degranulation of tumor cells leads to upregulation of CD107α in CTLs, which can be detected by anti-CD107α antibody.

[0148] Figure 31 A series of dot plots showing IFN-γ expression and degranulation of peptide-specific CTLs. CTLs were unstimulated or stimulated with KMS11 cells or OPM1 cells, as shown in the top panel. The peptide specificity of CTLs is shown in the left panel. The Y-axis represents CD107α expression, and the X-axis represents IFN-γ expression.

[0149] Figure 32 Schematic diagram of an assay for measuring the proliferation of peptide-specific CTLs in response to multiple myeloma cells. Peptide-specific CTLs were incubated with irradiated multiple myeloma cells for 6 or 8 days, and the proliferation of CTLs was measured by incorporation of CFSE.

[0150] Figure 33a A series of histograms showing the proliferative response of peptide-specific CTLs to myeloma cells on day 6. The peptide specificity of CTLs is shown in the left panel. The X-axis represents the decrease in CFSE staining, which is a direct measure of cell proliferation.

[0151] Figure 33b A series of histograms showing the proliferative response of peptide-specific CTLs to myeloma cells on day 8. The peptide specificity of CTLs is shown in the left panel. The X-axis represents the decrease in CFSE staining, which is a direct measure of cell proliferation.

[0152] Figure 34 A series of dot plots showing IL-2 production by peptide-specific CTLs in response to myeloma cells. The peptide specificity of the CTLs is shown in the left panel. The Y-axis represents IL-2 expression, and the X-axis represents CD8 expression. The boxed area represents the IL-2+CD8+ population.

[0153] Figure 35 A series of dot plots showing IFN-γ expression by peptide-specific CTLs from donor A in response to various colon cancer cell lines. As shown in the top panel, the CTLs were either unstimulated or stimulated with SW80, WiDr, or LS180 cells. The peptide specificity of the CTLs is shown in the left panel. The Y-axis represents IFN-γ expression, and the X-axis represents CD8 expression. The boxed area represents the IFN-γ+CD8+ population.

[0154] Figure 36 A series of dot plots showing IFN-γ expression and degranulation in peptide-specific CTLs from donor A in response to various colon cancer cell lines. As shown in the top panel, the CTLs were either unstimulated or stimulated with SW80, WiDr, or LS180 cells. The peptide specificity of the CTLs is shown in the left panel. The Y-axis represents CD107α expression, and the X-axis represents IFN-γ expression.

[0155] Figure 37 A series of dot plots showing IFN-γ expression by peptide-specific CTLs from donor B in response to various colon cancer cell lines. As shown in the top panel, the CTLs were either unstimulated or stimulated with SW80, WiDr, or LS180 cells. The peptide specificity of the CTLs is shown in the left panel. The Y-axis represents IFN-γ expression, and the X-axis represents CD8 expression. The boxed area represents the IFN-γ+CD8+ population.

[0156] Figure 38 A series of dot plots showing IFN-γ expression and degranulation in peptide-specific CTLs from group B. As shown in the top panel, the CTLs were either unstimulated or stimulated with SW80, WiDr, or LS180 cells. The peptide specificity of the CTLs is shown in the left panel. The Y-axis represents CD107α expression, and the X-axis represents IFN-γ expression.

[0157] Figure 39a A series of histograms showing the degranulation response of CD138 peptide-specific CTLs to SW480 tumor cells. As shown in the left panel, the CTLs and SW480 tumor cells were co-incubated at various cell:cell ratios, and the expression of CD107α, IFN-γ, and IL-2 was analyzed, as shown in the top panel.

[0158] Figure 39bA series of histograms showing the various responses of CD138 - peptide - specific CTLs to LS180 tumor cells. As shown in the left - hand figure, CTLs and LS180 tumor cells were co - incubated at various cell:cell ratios, and the expressions of CD107α, IFN - γ, and IL - 2 were analyzed, as shown in the top figure.

[0159] Figure 40a A series of dot plots showing the IFN - γ expression of peptide - specific CTLs from donor A in response to different cancer cell types. CTLs were incubated alone or co - incubated with SW480 colon cancer cells or KMS11 multiple myeloma cells, and IFN - γ expression was analyzed. The peptide - specificity of the CTLs is shown in the left - hand figure. The Y - axis represents IFN - γ expression, and the X - axis represents CD8 expression. The boxed area represents the IFN - γ+CD8+ population.

[0160] Figure 40b A series of dot plots showing the degranulation and IFN - γ expression of peptide - specific CTLs from donor A. CTLs were incubated alone or co - incubated with SW480 colon cancer cells or KMS11 multiple myeloma cells, and IFN - γ expression was analyzed. The peptide - specificity of the CTLs is shown in the left - hand figure. The Y - axis represents CD107α expression, and the X - axis represents IFN - γ expression.

[0161] Figure 41a A series of dot plots showing the IFN - γ expression of peptide - specific CTLs from donor B in response to different cancer cell types. CTLs were incubated alone or co - incubated with SW480 colon cancer cells or KMS11 multiple myeloma cells, and IFN - γ expression was analyzed. The peptide - specificity of the CTLs is shown in the left - hand figure. The Y - axis represents IFN - γ expression, and the X - axis represents CD8 expression. The boxed area represents the IFN - γ+CD8+ population.

[0162] Figure 41b A series of dot plots showing the degranulation and IFN - γ expression of peptide - specific CTLs from donor B in response to different cancer cell types. CTLs were incubated alone or co - incubated with SW480 colon cancer cells or KMS11 multiple myeloma cells, and IFN - γ expression was analyzed. The peptide - specificity of the CTLs is shown in the left - hand figure. The Y - axis represents CD107α expression, and the X - axis represents IFN - γ expression.

[0163] Figure 42A series of dot plots showing IFN-γ expression of peptide-specific CTLs from donor B in response to respective pancreatic cancer cell types. CTLs were incubated alone or co-incubated with 8902 cells, PL45 cells, or MiaPaca cells, and IFN-γ expression was analyzed. Peptide specificity of CTLs is shown in the left panel. The Y-axis represents IFN-γ expression, and the X-axis represents CD8 expression. The boxed area represents the IFN-γ+CD8+ population.

[0164] Figure 43 Histograms and dot plots showing different responses of CD138 peptide-specific CTLs to Panc1 pancreatic tumor cells. As shown in the left panel, CTL cells and Panc1 cells were co-incubated at a cell:cell ratio of 1:1 or 1:5, and the expression of CD107α, IFN-γ, and IL-2 was analyzed, as shown in the top panel. The Y-axis of the dot plot indicates CD107α expression, and the X-axis of the histograms and dot plots indicates IFN-γ or IL-2 expression, as shown below each figure.

[0165] Figure 44 A series of dot plots showing phenotypic changes in T cells (untreated T cells, central memory (CM) cells, effector cells, and effector memory (EM) CD8 + T cells) in CTLs induced by a mixture of heteroclitic unspliced XBP1 peptide and spliced XBP1 peptide. The Y-axis in the dot plots shows CCR7 expression, and the X-axis indicates CD45RP expression.

[0166] Figure 45 A series of bar graphs depicting the generation of central memory CD3+CD8+ T cells in three donors in response to a mixture of heteroclitic unspliced XBP1 peptide and spliced XBP1 peptide.

[0167] Figure 46 A series of bar graphs depicting the generation of effector memory CD3+CD8+ T cells in three donors in response to a mixture of heteroclitic unspliced XBP1 peptide and spliced XBP1 peptide.

[0168] Figure 47 A series of histograms showing the proliferative responses of XBP1 peptide mixture-specific CTLs to MB231 breast cancer cells. The responses were divided into proliferation in CD45RO- non-memory cells and CD45RO+ memory cells, and among the memory cells, a part was CD45RO+, CCR7+ central memory T cells, and a part was CD45RO+, CCR7- effector memory T cells.

[0169] Figure 48Are a series of histograms showing the proliferative responses of XBP1 peptide mixture-specific CTLs against LS180 colon cancer cells. The responses are divided into proliferation in CD45RO - non-memory cells and CD45RO + memory cells, and among the memory cells, a part is CD45RO +, CCR7 + central memory T cells, and a part is CD45RO +, CCR7 - effector memory T cells.

[0170] Figure 49 Are a series of histograms showing the proliferative responses of XBP1 peptide mixture-specific CTLs against Panc1 pancreatic cancer cells. The responses are divided into proliferation in CD45RO - non-memory cells and CD45RO + memory cells, and among the memory cells, a part is CD45RO +, CCR7 + central memory T cells, and a part is CD45RO +, CCR7 - effector memory T cells.

[0171] Figure 50 Are a series of histograms showing various responses of XBP1 peptide mixture-specific CTLs against various tumor cells (MB231, MCF7, LS180, SW480, Panc1, and PL45). IFN-γ expression of central memory T cells and effector memory T cells was analyzed.

[0172] Figure 51 Is a bar graph showing IFN-γ expression of effector memory T cells and central memory T cells of XBP1 mixture-specific CTLs against various tumor cells (MB231, MC7, LS180, SW480, Panc1, and PL45).

[0173] Figure 52 Are a series of histograms showing various responses of XBP1 peptide mixture-specific CTLs against various tumor cells (MB231, MCF7, LS180, SW480, Panc1, and PL45). IL-2 expression of central memory T cells and effector memory T cells was analyzed.

[0174] Figure 53 Depicts a bar graph showing IL-2 expression of effector memory T cells and central memory T cells of XBP1 mixture-specific CTLs against various tumor cells (MB231, MC7, LS180, SW480, Panc1, and PL45).

[0175] Figure 54 Are a series of histograms showing various responses of XBP1 peptide mixture-specific CTLs against various tumor cells (MB231, MCF7, LS180, SW480, Panc1, and PL45). Cytotoxicity of central memory T cells and effector memory T cells was analyzed.

[0176] Figure 55 Depicts a bar graph showing the cytotoxicity of effector memory T cells and central memory T cells of XBP1 cocktail-specific CTLs against various tumor cells (MB231, MC7, LS180, SW480, Panc1, and PL45).

[0177] Figure 56 Depicts a bar graph showing the Tbet expression of non-memory T cells and memory T cells of XBP1 cocktail-specific CTLs.

[0178] Figure 57 Depicts a bar graph showing the Tbet expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail-specific CTLs.

[0179] Figure 58 Depicts a bar graph showing the Tbet and IFN-γ expression of non-memory T cells and memory T cells of XBP1 cocktail-specific CTLs against various tumor cells (MB231, SW480, and Panc1).

[0180] Figure 59 Depicts a bar graph showing the Tbet and IFN-γ expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail-specific CTLs against MB231 breast cancer cells.

[0181] Figure 60 Depicts a bar graph showing the Tbet and IFN-γ expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail-specific CTLs against Panc1 pancreatic cancer cells.

[0182] Figure 61 Depicts a bar graph showing the Tbet and IFN-γ expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail-specific CTLs against SW480 colon cancer cells.

[0183] Figure 62 Depicts a bar graph showing the Eomes expression of non-memory T cells and memory T cells of XBP1 cocktail-specific CTLs.

[0184] Figure 63 Depicts a bar graph showing the Eomes expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 cocktail-specific CTLs.

[0185] Figure 64Depicts a bar graph showing the Eomes and IFN-γ expression of non-memory T cells and memory T cells of XBP1 mixture-specific CTLs against various tumor cells (MB231, SW480, and Panc1).

[0186] Figure 65 Depicts a bar graph showing the Eomes and IFN-γ expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against MB231 breast cancer cells.

[0187] Figure 66 Depicts a bar graph showing the Eomes and IFN-γ expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against Panc1 pancreatic cancer cells.

[0188] Figure 67 Depicts a bar graph showing the Eomes and IFN-γ expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against SW480 colon cancer cells.

[0189] Figure 68 Depicts a bar graph showing the granzyme B expression of non-memory T cells and memory T cells of XBP1 mixture-specific CTLs against various tumor cells (MB231, SW480, and Panc1).

[0190] Figure 69 Depicts a bar graph showing the granzyme B and IFN-γ expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against MB231 breast cancer cells.

[0191] Figure 70 Depicts a bar graph showing the granzyme B and IFN-γ expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against Panc1 pancreatic cancer cells.

[0192] Figure 71 Depicts a bar graph showing the granzyme B and IFN-γ expression of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against SW480 colon cancer cells.

[0193] Figure 72 Depicts a bar graph showing the number of non-memory T cells and memory T cells of XBP1 mixture-specific CTLs in the presence or absence of the adjuvant lenalidomide.

[0194] Figure 73 Depicts a bar graph showing the numbers of central memory T cells and effector memory T cells of XBP1 mixture-specific CTLs in the presence or absence of the adjuvant lenalidomide.

[0195] Figure 74 Depicts a bar graph showing the expressions of CD40L, CD69, and CD38 in XBP1 mixture-specific CTLs in the presence or absence of the adjuvant lenalidomide.

[0196] Figure 75 Depicts a bar graph showing the Tbet and IFN-γ expressions of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against MB231 breast cancer cells in the presence or absence of the adjuvant lenalidomide.

[0197] Figure 76 Depicts a bar graph showing the Eomes and IFN-γ expressions of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against MB231 breast cancer cells in the presence or absence of the adjuvant lenalidomide.

[0198] Figure 77 Depicts a bar graph showing the Tbet and IFN-γ expressions of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against Panc1 pancreatic cancer cells in the presence or absence of the adjuvant lenalidomide.

[0199] Figure 78 Depicts a bar graph showing the Eomes and IFN-γ expressions of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against Panc1 pancreatic cancer cells in the presence or absence of the adjuvant lenalidomide.

[0200] Figure 79 Depicts a bar graph showing the Tbet and IFN-γ expressions of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against SW480 colon cancer cells in the presence or absence of the adjuvant lenalidomide.

[0201] Figure 80 Depicts a bar graph showing the Eomes and IFN-γ expressions of untreated T cells, central memory cells, effector memory cells, and effector T cells of XBP1 mixture-specific CTLs against SW480 colon cancer cells in the presence or absence of the adjuvant lenalidomide.

[0202] Figure 81 Depicts a bar graph showing granzyme and IFN-γ expression of non-memory and memory T cells of XBP1 mixture-specific CTLs against MB231 breast cancer cells in the presence or absence of the adjuvant lenalidomide.

[0203] Figure 82 Depicts a bar graph showing granzyme and IFN-γ expression of non-memory and memory T cells of XBP1 mixture-specific CTLs against Panc1 pancreatic cancer cells in the presence or absence of the adjuvant lenalidomide.

[0204] Figure 83 Depicts a bar graph showing granzyme and IFN-γ expression of non-memory and memory T cells of XBP1 mixture-specific CTLs against SW480 colon cancer cells in the presence or absence of the adjuvant lenalidomide. DETAILED DESCRIPTION OF THE INVENTION

[0206] The present disclosure features immunogenic XBP1-derived peptides, CD138-derived peptides, and CS-1-derived peptides (and pharmaceutical compositions thereof) that can be used, for example, to induce an immune response (e.g., stimulate a CTL response) in a subject or to stimulate the production of antibodies. The peptides can be used in a variety of applications, such as methods for inducing an immune response, methods for producing antibodies, and methods for treating cancer (e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia (e.g., AML or CML), and plasma cell disorders such as multiple myeloma or Waldenstrom macroglobulinemia) or pre-cancerous disorders (e.g., smoldering multiple myeloma). The peptides can also be included in MHC molecule multimer compositions and used, for example, in methods for detecting T cells in a cell population.

[0207] The following provides a detailed description of the peptides and exemplary methods for producing and using the peptides.

[0208] Peptide

[0209] Group A peptides. The present disclosure features isolated peptides (“Group A peptides”) that comprise an amino acid sequence having sufficient identity or being identical to any one of SEQ ID NOs: 1-18 depicted in Table 1.

[0210] Table 1. Examples of Peptides in Group A

[0211] Origin Protein Amino Acid Position Amino Acid Sequence SEQ ID NO: Unspliced XBP1 118-126 LLREKTHGL 1 Unspliced XBP1 185-193 NISPWILAV 2 Unspliced XBP1 190-198 ILAVLTLQI 3 Unspliced XBP1 193-201 VLTLQIQSL 4 Unspliced XBP1 111-119 KLLLENQLL 5 Unspliced 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

[0212] The bolded residues represent amino acid changes from the corresponding wild-type amino acid sequence.

[0213] Preferably, the isolated peptides from Group A are 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 amino acids in length, 9 - 25 amino acids in length, 9 - 20 amino acids in length, 9 - 15 amino acids in length), and comprise an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or identical to the amino acid sequences of SEQ ID NOs: 1 - 18. Other preferred peptides are 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 amino acids in length, 9 - 25 amino acids in length, 9 - 20 amino acids in length, 9 - 15 amino acids in length), and comprise the amino acid sequences of SEQ ID NOs: 1 - 18, or the amino acid sequences of SEQ ID NOs: 1 - 18 with one, two, three or four substituted amino acids. The substitutions can be conservative substitutions or non-conservative substitutions.

[0214] The "unspliced XBP1" peptides from Group A include those described in Table 1 and refer to peptides 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) contiguous amino acids from the unspliced form of the human XBP1 protein, the unspliced form of the human XBP1 protein having 261 amino acids and the following sequence:

[0215] MVVVAAAPNPADGTPKVLLLSGQPASAAGAPAGQALPLMVPAQRGASPEAASGGLPQARKRQRLTHLSPEEKALRRKLKNRVAAQTARDRKKARMSELEQQVVDLEEENQKLLLENQLLREKTHGLVVENQELRQRLGMDALVAEEEAEAKGNEVRPVAGSAESAALRLRAPLQQVQAQLSPLQNISPWILAVLTLQIQSLISCWAFWTTWTQSCSSNALPQSLPAWRSSQRSTQKDPVPYQPPFLCQWGRHQPSWKPLMN (SEQ ID NO:19; Genbank accession number NP_005071), and peptides having no more than one, two, three, four, or five amino acid substitutions (e.g., conservative substitutions) derived from the amino acid sequence of SEQ ID NO:19. Amino acid positions referred to in Table 1 are based on SEQ ID NO:19.

[0216] "Spliced XBP1" peptides from Group A include those peptides described in Table 1 and refer to peptides 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) contiguous amino acids from the spliced form of the human XBP1 protein (XBP1splice), which has 376 amino acids and the following sequence:

[0217] MVVVAAAPNPADGTPKVLLLSGQPASAAGAPAGQALPLMVPAQRGASPEAASGGLPQARKRQRLTHLSPEEKALRRKLKNRVAAQTARDRKKARMSELEQQVVDLEEENQKLLLENQLLREKTHGLVVENQELRQRLGMDALVAEEEAEAKGNEVRPVAGSAESAAGAGPVVTPPEHLPMDSGGIDSSDSESDILLGILDNLDPVMFFKCPSPEPASLEELPEVYPEGPSSLPASLSLSVGTSSAKLEAINELIRFDHIYTKPLVLEIPSETESQANVVVKIEEAPLSPSENDHPEFIVSVKEEPVEDDLVPELGISNLLSSSHCPKPSSCLLDAYSDCGYGGSLSPFSDMSSLLGVNHSWEDTFANELFPQLISV (SEQ ID NO:20; Genbank accession number NP_001073007), and peptides having an amino acid sequence derived from the amino acid sequence of SEQ ID NO:20 and having no more than one, two, three, four, or five amino acid substitutions (e.g., conservative substitutions). The amino acid positions referred to in Table 1 are based on SEQ ID NO:20.

[0218] The "CD138" peptides from Group A include those depicted in Table 1 and refer to peptides 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) contiguous amino acids from the human CD138 protein, which human CD138 protein has 310 amino acids and the following sequence:

[0219] MRRAALWLWLCALALSLQPALPQIVATNLPPEDQDGSGDDSDNFSGSGAGALQDITLSQQTPSTWKDTQLLTAIPTSPEPTGLEATAASTSTLPAGEGPKEGEAVVLPEVEPGLTAREQEATPRPRETTQLPTTHQASTTTATTAQEPATSHPHRDMQPGHHETSTPAGPSQADLHTPHTEDGGPSATERAAEDGASSQLPAAEGSGEQDFTFETSGENTAVVAVEPDRRNQSPVDQGATGASQGLLDRKEVLGGVIAGGLVGLIFAVCLVGFMLYRMKKKDEGSYSLEEPKQANGGAYQKPTKQEEFYA (SEQ ID NO:21; GenBank accession number NP_002988), and peptides having an amino acid sequence derived from the amino acid sequence of SEQ ID NO:21 and having no more than one, two, three, four, or five amino acid substitutions (e.g., conservative substitutions). Amino acid positions referred to in Table 1 are based on SEQ ID NO:21.

[0220] The "CS-1" peptides from Group A include those described in Table 1 and refer to peptides 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) contiguous amino acids from the human CS-1 protein, the human CS-1 protein having 335 amino acids and the following sequence:

[0221] MAGSPTCLTLIYILWQLTGSAASGPVKELVGSVGGAVTFPLKSKVKQVDSIVWTFNTTPLVTIQPEGGTIIVTQNRNRERVDFPDGGYSLKLSKLKKNDSGIYYVGIYSSSLQQPSTQEYVLHVYEHLSKPKVTMGLQSNKNGTCVTNLTCCMEHGEEDVIYTWKALGQAANESHNGSILPISWRWGESDMTFICVARNPVSRNFSSPILARKLCEGAADDPDSSMVLLCLLLVPLLLSLFVLGLFLWFLKRERQEEYIEEKKRVDICRETPNICPHSGENTEYDTIPHTNRTILKEDPANTVYSTVEIPKKMENPHSLLTMPDTPRLFAYENVI (SEQ ID NO:22; Genbank accession number NP_067004), and peptides having an amino acid sequence derived from SEQ ID NO:22 and having no more than one, two, three, four, or five amino acid substitutions (e.g., conservative substitutions). The amino acid positions referred to in Table 1 are based on SEQ ID NO:22.

[0222] Group B peptides. The present disclosure features isolated peptides (“Group B peptides”) comprising an amino acid sequence having sufficient identity to or identical to any of SEQ ID NOs: 29 - 50 depicted in Table 2.

[0223] Table 2. Examples of Peptides in Group B

[0224]

[0225]

[0226] Preferably, the isolated peptides from group B have a length of at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, or 35 amino acids (e.g., between 9 and 35 amino acids in length, e.g., 9 - 30 amino acids in length, 9 - 25 amino acids in length, 9 - 20 amino acids in length, 9 - 15 amino acids in length), and comprise an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or being identical to the amino acid sequences of SEQ ID NOs: 29 - 50. Other preferred peptide lengths can be at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, or 35 amino acids (e.g., between 9 and 35 amino acids in length, e.g., 9 - 30 amino acids in length, 9 - 25 amino acids in length, 9 - 20 amino acids in length, 9 - 15 amino acids in length), and comprise the amino acid sequences of SEQ ID NOs: 29 - 50, or the amino acid sequences of SEQ ID NOs: 29 - 50 and having an amino acid sequence with one, two, three, or four substituted amino acids. The substitution can be a conservative substitution or a non - conservative substitution.

[0227] "Unspliced XBP1" from group B includes those peptides described in Table 2 and refers to peptides 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 unspliced form of the human XBP1 protein, the unspliced form of the human XBP1 protein having 261 amino acids and the amino acid sequence of SEQ ID NO: 19, and peptides derived from the amino acid sequence of SEQ ID NO: 19 and having no more than one, two, three, four, five amino acid substitutions (e.g., conservative substitutions). Unspliced 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 of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids at the N - terminus and / or C - terminus of any one of SEQ ID NOs: 29 and 33 - 37. The amino acid positions referred to in Table 2 are based on SEQ ID NO: 19.

[0228] The "spliced XBP1" peptides from Group B include those described in Table 2 and refer to peptides 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) contiguous amino acids from the spliced form of the human XBP1 protein (XBP1 splicing), wherein the spliced form of the human XBP1 protein has 376 amino acids and the amino acid sequence of SEQ ID NO:20, as well as peptides derived from the amino acid sequence of SEQ ID NO:20 and having no more than one, two, three, four, or five amino acid substitutions (e.g., conservative substitutions). The spliced XBP1 peptides from Group B include peptides having the amino acid sequence of SEQ ID NO:20, which amino acid sequence contains part or all of any one of SEQ ID NO:30, 38, and 39, e.g., a sequence of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids at the N-terminus and / or C-terminus of any one of SEQ ID NO:30, 38, and 39. The amino acid positions mentioned in Table 2 are based on SEQ ID NO:20.

[0229] The "CD138" peptides from Group B include those depicted in Table 2 and refer to peptides 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, the human CD138 protein having 310 amino acids and the amino acid sequence of SEQ ID NO:21, and peptides derived from the amino acid sequence of SEQ ID NO:21 and having no more than one, two, three, four, or five amino acid substitutions (e.g., conservative substitutions). The CD138 peptides from Group B include peptides having an amino acid sequence from SEQ ID NO:21 that includes part or all of any one of SEQ ID NO:31 and 40 - 45, e.g., a sequence of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids at the N-terminus and / or C-terminus that includes SEQ ID NO:31 and any one of 46 - 50. The amino acid positions mentioned in Table 2 are based on SEQ ID NO:21.

[0230] The "CS-1" peptides from Group B include those peptides described in Table 2 and refer to peptides 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, the human CS-1 protein having 335 amino acids and the amino acid sequence of SEQ ID NO:22, and peptides derived from the amino acid sequence of SEQ ID NO:22 and having no more than one, two, three, four, five amino acid substitutions (e.g., conservative substitutions). The CS1 peptides from Group B include peptides having an amino acid sequence from SEQ ID NO:22, the amino acid sequence comprising part or all of any one of SEQ ID NO:32 and 46 - 50, e.g., a sequence of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids at the N-terminus and / or C-terminus of any one of SEQ ID NO:32 and 46 - 50. The amino acid positions mentioned in Table 2 are based on SEQ ID NO:22.

[0231] Group C peptides. The present disclosure features isolated peptides ("Group C peptides") that comprise an amino acid sequence having sufficient identity or being identical to any one of SEQ ID NOs: 51 - 536 as described in Table 3.

[0232] Table 3. Examples of Peptides in Group C

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242]

[0243] Preferably, the isolated peptides from Group C have a length of at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, or 35 amino acids (e.g., between 9 and 35 amino acids in length, e.g., 9 - 30 amino acids in length, 9 - 25 amino acids in length, 9 - 20 amino acids in length, 9 - 15 amino acids in length), and comprise an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or being identical to the amino acid sequences of SEQ ID NOs: 51 - 536. Other peptide lengths can be 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, or 35 amino acids (e.g., between 9 and 35 amino acids in length, e.g., 9 - 30 amino acids in length, 9 - 25 amino acids in length, 9 - 20 amino acids in length, 9 - 15 amino acids in length), and comprise the amino acid sequences of SEQ ID NOs: 51 - 536, or the amino acid sequences of SEQ ID NOs: 51 - 536 with one, two, three, or four amino acid substitutions. The substitutions can be conservative substitutions or non - conservative substitutions.

[0244] "Unspliced XBP1" from Group C includes those peptides described in Table 3, and refers to peptides 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 unspliced form of human XBP1, the unspliced form of the human XBP1 protein having 261 amino acids and the amino acid sequence of SEQ ID NO: 19, and peptides derived from the amino acid sequence of SEQ ID NO: 19 with no more than one, two, three, four, five amino acid substitutions (e.g., conservative substitutions). Unspliced XBP1 peptides from Group C include peptides having an amino acid sequence from SEQ ID NO: 19, the amino acid sequence comprising 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.

[0245] The "CD138" peptides from Group C include those depicted in Table 3 and refer to peptides 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) contiguous amino acids from the human CD138 protein, which human CD138 protein has 310 amino acids and the amino acid sequence of SEQ ID NO:21, as well as peptides derived from the amino acid sequence of SEQ ID NO:21 and having no more than one, two, three, four, five amino acid substitutions (e.g., conservative substitutions). The CD138 peptides from Group C include peptides having an amino acid sequence from SEQ ID NO:21, which amino acid sequence comprises part or all of SEQ ID NO:207 - 371. The amino acid positions mentioned in Table 3 are based on SEQ ID NO:21.

[0246] The "CS-1" peptides from Group C include those described in Table 3 and refer to peptides 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) contiguous amino acids from the human CS-1 protein, which human CS-1 protein has 335 amino acids and the amino acid sequence of SEQ ID NO:22, as well as peptides derived from the amino acid sequence of SEQ ID NO:22 and having no more than one, two, three, four, five amino acid substitutions (e.g., conservative substitutions). The CS1 peptides from Group C include peptides having an amino acid sequence from SEQ ID NO:22, which amino acid sequence comprises part or all of SEQ ID NO:372 - 536. The amino acid positions mentioned in Table 3 are based on SEQ ID NO:22.

[0247] Peptide Overview

[0248] The residue numbers of the N-terminal and C-terminal amino acids of the peptide are typically utilized (e.g., XBP1 118-126)When referring to the peptides described herein, it is as if the relevant sequences occur in the wild-type, full-length, mature human proteins having SEQ ID NOs: 19-22. These peptides will generally have the same sequence as the corresponding segments of the wild-type, full-length, mature proteins having SEQ ID NOs: 19-22. However, it should be understood that the terms "unspliced XBP1 peptide" (e.g., an unspliced XBP1 peptide having the following amino acid positions: 118-136, 185-193, 186-194, 190-198, 193-200 or 111-119), "spliced XBP1 peptide" (e.g., a spliced XBP1 peptide having the following amino acid positions: 197-205, 194-202, 224-232, 368-376), "CD138 peptide" (e.g., a CD138 peptide having the following amino acid positions: 256-264, 265-273, 260-268, 5-13 or 7-15), and CS1 peptide (e.g., a CS1 peptide having the following amino acid positions: 236-245, 240-248, 239-247, 232-240 or 9-17) can be fragments of the XBP1 unspliced peptide, XBP1 spliced peptide, CD138 or CS-1 polypeptide (respectively) of non-human species. As will be understood by those skilled in the art, the number of N-terminal and C-terminal amino acids of the peptide fragments of the non-human polypeptides need not be the same as those in the corresponding peptide fragments of the human polypeptides. Also, the length and / or amino acids of the peptide fragments of the non-human polypeptides need not be the same as those in the corresponding peptide fragments of the human polypeptides. Those skilled in the art will know how to establish the N- and C-terminal amino acids, length, and amino acid sequence of the peptides derived from non-human unspliced XBP1 polypeptides, spliced XBP1 polypeptides, CD138 polypeptides, and CS-1 polypeptides. A useful method for doing so is sequence alignment, and, in particular, maximum homology sequence alignment.

[0249] The percent identity between two peptide sequences (e.g., the peptides 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 (European Bioinformatics Institute (EMBL-EBI)), BLAST-Protein (National Center for Biotechnology Information (NCBI), National Institutes of Health, USA), and PSAlign (University of Texas A&M; Sze et al. (2006) Journal of Computational Biology 13:309-319).

[0250] The present disclosure also discloses variants of the human and non-human peptides described above. Variants of the human and non-human peptides described herein may include peptide forms having: (i) no more than 4 (e.g., 3, 2, or 1) amino acid substitutions (e.g., conservative or non-conservative substitutions); (ii) terminal deletions or internal deletions; or (iii) terminal additions or internal additions, all of which are detailed below.

[0251] The present disclosure is also characterized by a peptide comprising, consisting of, or consisting essentially of any one of SEQ ID NOs: 1-18 and 29-536 (depicted in Tables 1-3), but having an amino acid sequence with no more than four (e.g., no more than three, no more than two, or no more than one) substitutions. The substitutions can be, for example, conservative or non-conservative (as described above).

[0252] Conservative substitutions include substitutions from 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. Non-polar hydrophobic amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine. Polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine. Positively charged (basic) amino acids include arginine, lysine, and histidine. Negatively charged (acidic) amino acids include aspartic acid and glutamic acid. Any substitution of a member of the above-mentioned polar, basic, or acidic group by another member of the same group can be regarded as a conservative substitution. In contrast, a non-conservative substitution is the substitution of one amino acid by another amino acid having dissimilar properties.

[0253] 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 peptide are not substituted. In some embodiments, one or more of positions three, four, five, six, seven, and eight of any peptide are the same as the amino acids of the peptides in Tables 1-3.

[0254] The disclosure is further characterized by a fusion protein comprising: a first amino acid sequence of a peptide described herein (e.g., an unspliced 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 heterologous to the first amino acid sequence.

[0255] The second heterologous amino acid sequence of the peptide generally does not (and is selected so as not to) adversely affect the production of the immunogenic peptides of any of SEQ ID NOs: 1-18 and 29-536 in cells. It is expected that the cellular machinery removes any additional sequences in the peptide to generate the immunogenic peptides of any of SEQ ID NOs: 1-18 and 29-536, which are presented by class I or class II MHC molecules to stimulate an immune response against XBP1-expressing cancer cells, CD138-expressing cancer cells, or CS-1-expressing cancer cells.

[0256] 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 sequences that flank the first amino acid sequence as it exists in nature. For example, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 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, 9, 8, 7, 6, 5, 4, 3, 2, or 1) carboxy-terminal and / or amino-terminal amino acids that flank LLREKTHGL (SEQ ID NO:1) in human XBP1 are considered not to be heterologous to SEQ ID NO:1. It is understood that a fusion protein containing the first amino acid sequence may not exist in nature at all, the first amino acid sequence of which is less than 100% identical to the amino acid sequence of any of SEQ ID NOs: 1-18 and 29-536, or contains one to four conservative substitutions in the amino acid sequence of any of SEQ ID NOs: 1-18 and 29-536.

[0257] 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 acids that flank the first amino acid sequence as it exists in nature. For example, two amino acids that flank LLREKTHGL (SEQ ID NO:1) in human XBP1 are considered not to be heterologous to SEQ ID NO:1.

[0258] Heterologous sequences can be, for example, sequences for purifying recombinant proteins (e.g., FLAG, polyhistidine (e.g., hexahistidine) (SEQ ID NO: 544), hemagglutinin (HA), glutathione-S-transferase (GST), or maltose binding protein (MBP)). Heterologous sequences can also be proteins used as diagnostic markers or detectable markers, such as luciferase, green fluorescent protein (GFP), or chloramphenicol acetyltransferase (CAT). In some embodiments, the fusion protein can include a signal sequence from another protein, such as the KDEL (SEQ ID NO: 23) sequence or any other described herein. In some embodiments, the fusion protein can include 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 include a useful therapeutic polypeptide or immunostimulatory polypeptide (e.g., all or part of a T helper epitope (e.g., the PADRE epitope or the tetanus toxoid universal T helper cell epitope) or a cytokine or chemokine) and / or a carrier (e.g., KLH), for example, for eliciting an immune response (e.g., for antibody production). In some embodiments, the fusion protein can include one or more linkers, such as linkers containing peptide sequences (see below). The fusion protein can also include a targeting polypeptide. Heterologous sequences can be of different lengths and, in some cases, can be sequences that are longer than the first amino acid sequence to which the heterologous amino acids are attached. It is understood that a fusion protein comprising a first amino acid sequence and a second amino acid sequence heterologous to the first amino acid sequence will not correspond in sequence to a naturally occurring protein.

[0259] As used herein, a targeting polypeptide is a polypeptide that targets the moiety to which it is attached (e.g., the first amino acid sequence) to a specific tissue (e.g., targets lymph nodes) or cell (e.g., targets antigen-presenting cells or other immune cells), or which, when in vitro, targets a specific isolated molecule or molecular complex. A targeting polypeptide can be, for example, an antibody (immunoglobulin) or an antigen-binding fragment thereof or a ligand of a cell surface receptor. An antibody (or an antigen-binding fragment thereof) can be, for example, a monoclonal antibody, a polyclonal antibody, a humanized antibody, a fully human antibody, a single-chain antibody, a chimeric antibody, or a Fab fragment, F(ab’) 2Fragment, Fab’ fragment, Fv fragment or scFv fragment. Antibody fragments comprising an Fc region or an Fc region (with or without an antigen-binding region) can also be used to target reagents to Fc receptor-expressing cells (e.g., antigen-presenting cells such as cross-presenting dendritic cells, macrophages, monocytes or B cells). Ligands for cell surface receptors can be, for example, chemokines, cytokines (e.g., interleukin 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16), or death receptor ligands (e.g., FasL or TNFα).

[0260] In some embodiments, the heterologous sequence can be, for example, a “translocation sequence” (e.g., endoplasmic reticulum or Golgi apparatus organelles) for delivering a peptide to a cell or a specific part of a cell. Translocation sequences can include, for example, membrane translocation sequences, trafficking sequences, antennapedia sequences, cyclic integrin-binding peptides and Tat-mediated peptides, or modified forms thereof.

[0261] A linker (e.g., a linker peptide) can directly or indirectly link a first amino acid sequence to one or more heterologous amino acid sequences. For example, a linker can link a first amino acid sequence to a second amino acid sequence. The linker peptide can be, or comprise, for example, a fragment of amino acids, wherein at least four to six amino acids are glycine. (See, e.g., Mancebo et al. (1990) Mol. Cell. Biol. 10:2492-2502). The linker peptide can also be, or comprise, six or more (e.g., seven, eight, nine, 10, 11 or 12 or more) histidine residues. The linker peptide can be, or comprise, at least one (e.g., one, two, three, four, five, six, seven or eight or more) protease cleavage site. The protease site can be, for example, an insulin, chymotrypsin or factor Xa cleavage site. The protease site can be useful, for example, to separate the first amino acid sequence from the heterologous sequence. For example, after expression and purification of a fusion protein comprising a first amino acid sequence linked to a polyhistidine sequence (used for purification in this case), the polyhistidine sequence can be removed from the first amino acid sequence by contacting the fusion protein with insulin.

[0262] The first amino acid sequence and the second amino acid sequence can be combined with each other in a variety of ways. As used herein, "bind to" in the context of an interaction between two or more atomic or molecular units includes any covalent or non-covalent bonding, or physical mixture, of two or more atomic or molecular units (e.g., the first amino acid sequence and the second amino acid sequence). The chemical nature of a covalent bond (two atoms sharing two or more pairs of valence electrons) is known in the art and includes, for example, disulfide bonds or peptide bonds. A non-covalent bond is a chemical bond between atoms or molecules that does not involve the sharing of valence electron pairs. For example, non-covalent interactions include, for example, hydrophobic interactions, hydrogen bond interactions, ionic bonding, van der Waals bonding, or dipole-dipole interactions. Examples of such non-covalent interactions include antibody-antigen complex or binding pair interactions (the interaction between the first and second members of a binding pair such as the interaction between streptavidin and biotin). It is understood that the term "bind to" (e.g., in the case of the first amino acid sequence and the second amino acid sequence) coextends with the term "comprise".

[0263] In some embodiments, the first amino acid sequence and the second amino acid sequence can be encoded by a single nucleic acid sequence (and expressed therefrom as a fusion protein). In some instances, the first amino acid sequence and the 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 "Nucleic Acids and Methods for Producing Peptides" below).

[0264] When expressed or produced separately, any number of known chemical cross-linking linkers can be utilized to cross-link the first and second amino acid sequences together. Examples of such chemical cross-linking linkers are those that link two amino acid residues via a bond including a "hindered" disulfide bond. In these bonds, the disulfide bond in the cross-linking unit is protected by reduction (of the hindered groups on either side of the disulfide bond), which can be carried out, for example, by the action of reduced glutathione or an enzyme disulfide reductase. A suitable chemical cross-linker, 4-succinimidyloxycarbonyl-α-methyl-α-(2-pyridyldithio)toluene (SMPT), forms the said bond between two amino acid sequences using a terminal lysine on one of the amino acid sequences and a terminal cysteine on another amino acid. Heterobifunctional reagents cross-link via different coupling moieties on each amino acid sequence. In this way, the resulting "dimer" will be a heterodimer (a peptide containing the first and second amino acid sequences) rather than a homodimer or a mixture of homodimers and heterodimers (e.g., two first amino acid sequences or two second amino acid sequences). Thus, the coupling moiety on the first amino acid sequence can be a cysteine residue and on the other amino acid a lysine residue. Other useful cross-linkers include, but are not limited to, those that link two amino groups (e.g., N-5-azido-2-nitrobenzoic acid succinimidyl ester), two sulfhydryl groups (e.g., 1,4-di-maleimidobutane), an amino group and a sulfhydryl group (e.g., m-maleimidobenzoyl-N-hydroxysuccinimide ester), an amino and a carboxyl group (e.g., 4-[p-azidosalicylamido]butylamine), and chemicals that link the amino and guanadium groups present in the side chain of arginine (e.g., p-azidophenylglyoxal monohydrate).

[0265] The coupling moieties will preferably be at the termini (C or N) of each amino acid sequence. As shown above, they can be cysteine residues on each amino acid sequence or a cysteine on one amino acid sequence and a lysine on the other amino acid sequence. Where they are two cysteine residues, cross-linking can be subject to, for example, the influence of exposing the amino acid sequence to oxidative conditions.

[0266] The fusion protein can contain the first and second amino acid sequences or the fusion protein can contain more than one (e.g., two, three, four, five, six, seven, eight or more) additional heterologous amino acid sequences. The additional heterologous amino acid sequences can flank or be linked to the amino terminus and / or carboxyl terminus of the first amino acid sequence.

[0267] In cases where more than two amino acid sequences are to be joined, at least one of the amino acid sequences may have more than one cross-linking moiety. For example, the first amino acid sequence may have cross-linking moieties at the amino terminus and the carboxyl terminus. The polymer may be interpreted as being "sequentially". Thus, each amino acid sequence is joined to the next such that the terminal amino acid sequences in the chain have only one residue participating in the inter-domain (inter-agent) bonding, while the "internal" amino acid sequences have two moieties participating in the inter-domain bonding. Optionally, one amino acid sequence (such as the first amino acid sequence) may be joined to a plurality (e.g., 2, 3, 4, or 5) of other amino acid sequences.

[0268] Another feature is a peptide composition comprising: a first component and a second component, wherein the first component is a peptide as described herein. The second component may be, for example, 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, the peptide composition may comprise 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.

[0269] It is understood that in some embodiments, the peptides described herein may have up to 200 (e.g., 1, 2, 3, 4, 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, 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) heterologous amino acids at the amino terminus and / or the carboxyl terminus.

[0270] The peptides described herein can bind to major histocompatibility complex (MHC) molecules (e.g., MHC class I molecules or MHC class II molecules). "Major histocompatibility complex" or "MHC" is a cluster of genes that plays a role in regulating cell interactions responsible for physiological immune responses. In humans, the MHC is known as the HLA complex (see, e.g., Paul et al., FUNDAMENTAL IMMUNOLOGY, Third Edition, Raven Press, New York, (1993) and Stites, et al., IMMUNOLOGY, Eighth Edition, Lange Publishing, Los Altos, Calif. (1994))

[0271] As used herein, "HLA supertype or family" refers to a group of HLA molecules grouped based on shared peptide-binding specificities. HLA class I molecules that share a certain degree of similar binding affinity for peptides containing 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 represents a specific HLA type) are synonymous. Types of HLA class I molecules include, for example, HLA-A1, HLA-A2, HLA-A3, HLA-A24, HLA-B7, HLA-B27, HLA-B44, HLA-B58, or HLA-B62. The HLA molecules are described in detail in U.S. Patent No. 7,026,443, the entire content of which is incorporated herein by reference in its entirety.

[0272] Peptides can bind to MHC molecules with high affinity or moderate affinity. As used herein, "high affinity" binding of a peptide to an HLA class I molecule is defined as binding with a dissociation constant (K D ) of less than 50 (e.g., 45, 40, 35, 30, 25, 20, 15, 10, 5, 1, 0.5, 0.1, or less than 0.5) nM. "Moderate affinity" is when the peptide binds to an HLA class I molecule with a K 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) Dbinding. A peptide is defined as having "high affinity" for an HLA class II molecule if it binds with a K 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 D binding. "Moderate affinity" of a peptide for an HLA class II molecule is with a K between approximately 100 and approximately 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) D binding. Methods for determining the binding affinity of a peptide and an MHC are known in the art and are set forth in the appended examples. Suitable methods are also described in, for example, U.S. Patent No. 7,026,443.

[0273] The peptides described herein can also bind to MHC molecules and be recognized by antigen-specific T cell receptors on T cells. A variety of suitable methods can be used to determine whether a peptide that binds to 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 in vitro with a test peptide in the presence of antigen-presenting cells for several weeks. During this time, T cells specific for the peptide become activated and can be detected using, for example, a proliferation assay (carboxyfluorescein succinimidyl ester (CFSE) assay or 3 H-thymidine assay), a limiting dilution assay, a cytotoxicity assay (e.g., calcein release assay) or a cytokine (e.g., IFNγ), lymphokine assay or 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 entire contents of each of which are incorporated herein by reference in their entirety). Suitable in vivo methods involve immunizing HLA transgenic mice, wherein the peptide in adjuvant is administered subcutaneously to the HLA transgenic mice, and several weeks after immunization, spleen cells are removed and cultured in vitro in the presence of the test peptide for about one week, and using, for example, 51 Cr release assays (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 contents of each of which are incorporated herein by reference in their entirety) to detect peptide-specific T cells. Suitable methods are listed in the Examples below. For example, activation of T cells by a peptide (in the context of MHC molecules) can be by IFN-γ cytokine production, CD107α degranulation, or calcein release cytotoxicity assays (see, e.g., Examples 13 and 14).

[0274] In addition, direct quantification of antigen-specific T cells can be performed by staining T cells with MHC complexes labeled with detectable labels, such as any of the MHC molecule multimer compositions described herein (see below) or HLA-I tetramers (e.g., 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 herein by reference in their entirety).

[0275] In some embodiments, a peptide can be modified (e.g., an amino acid of the peptide can be replaced) to modulate (e.g., increase or decrease) one or more properties of the peptide. For example, one or more (e.g., two, three, or four) amino acids of a peptide depicted in Table 1 can be replaced to increase the peptide's affinity for an MHC molecule. In some embodiments, an amino acid of a peptide described herein (e.g., an amino acid residue of a T cell receptor contacting peptide) can be modified to enhance the binding interaction between a T cell receptor and the peptide (in the context of an MHC molecule). The modified peptide is generally referred to as an "altered peptide ligand" (see, e.g., Kalergis et al. (2000) J Immunol. 165(1):280; Conlon et al. (2002) Science 1801; and International Publication No. WO02070003, the disclosures of each of which are incorporated by reference in their entirety).

[0276] Suitable methods for modifying peptides and determining the effect of the modification are set forth in the appended examples and described, for example, in Collins et al. (Immunlogical Reviews (1998) 163:151-160, the disclosures of each of which are incorporated by reference in their entirety).

[0277] Nucleic Acids and Methods for Producing Peptides

[0278] The present disclosure features nucleic acid sequences (and nucleic acid vectors containing the nucleic acid sequences), and methods for producing one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of any of the peptides described herein (or fusion proteins described herein). The methods can include the steps of: optionally, providing a nucleic acid vector comprising a nucleic acid sequence containing one or more of any of the nucleic acid sequences encoding the peptides described herein (or fusion proteins described herein), the nucleic acid sequence being operably linked to an expression control sequence, and culturing the cells under conditions permitting the expression of the peptide (or fusion protein). The methods can further include the step of isolating one or more peptides (or proteins) from the cells or from the culture medium of the cultured cells.

[0279] Suitable methods for constructing nucleic acid sequences and vectors for the recombinant expression of one or more of the peptides (or fusion proteins) described herein are well known to those skilled in the art and are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Volumes 1, 2, and 3, Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York, USA, Nov. 1989, the disclosure of which is incorporated herein by reference in its entirety. The nucleic acids and vectors can be used to express the peptides (or fusion proteins) in a variety of host cells including, for example, bacteria, yeast, or mammalian cells. The nucleic acids and vectors can also be used, for example, in in vivo and ex vivo methods as described below.

[0280] A peptide coding sequence (or fusion protein coding sequence) can be operably linked to a promoter and / or enhancer element that directs the expression of the peptide (or fusion protein) encoded by the nucleic acid. Enhancers provide expression specificity in terms of time, location, and level. Unlike promoters, enhancers can function when located at different distances from the transcription start site (if a promoter is present). Enhancers can also be located downstream of the transcription start site or within exons of the relevant gene. For the coding sequence to be under the control of a promoter, the translation start site of the translation reading frame of the peptide needs to be placed between one nucleotide and 50 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 λ, the regulatory region of fd coat protein, the promoter of 3-phosphoglycerate kinase, the promoter of acid phosphatase, and the promoter of yeast α mating factor, the adenovirus E1b minimal promoter, or the thymidine kinase minimal promoter.

[0281] A peptide coding sequence, a fusion protein coding sequence, or a vector comprising said sequence may comprise a leader sequence encoding a signal peptide. The leader sequence may be at the 5' end of the sequence encoding one or more of the peptides or fusion proteins described herein. The signal peptide may be immediately adjacent to the N-terminus of a given peptide (or fusion protein) or may be separated from said given protein 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 is typically cleaved from the peptide (or fusion protein) prior to secretion (unless the signal peptide directs the insertion of a transmembrane protein), directs the peptide (or fusion protein) to which it is attached into the lumen of the endoplasmic reticulum (ER) of the host cell during translation, and then secretes the peptide (or fusion protein) via secretory vesicles into the environment of the host cell. Useful signal peptides include, for example, the native leader sequences of cytokines or even factors, KDEL (SEQ ID NO:23), or any signal sequence described, for example, in U.S. Patent No. 5,827,516, the disclosure of which is incorporated herein by reference in its entirety.

[0282] In some embodiments, the 5' end of the peptide coding sequence (or fusion protein coding sequence) may comprise a non-native ATG "start sequence". That is, for example, an ATG sequence may be added to the nucleic acid encoding the peptide (or fusion protein) to ensure proper transcription and translation of the peptide (or fusion protein). While the leader sequence typically includes an ATG start sequence, in embodiments where it does not include an ATG start sequence, an ATG sequence may be added to the 5' end of the nucleic acid encoding the leader sequence.

[0283] Suitable methods for constructing peptide coding sequences and expression vectors are known to those of skill in the art and are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual Second Edition vol. 1, 2 and 3. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York, USA, Nov. 1989, the entire contents of which are incorporated herein by reference in its entirety.

[0284] A variety of methods can be used to introduce a recombinant vector into a cell, and the method can (at least in part) depend 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, for example, the spheroplast technique or the whole-cell lithium chloride yeast transformation method (see, for example, U.S. Patent 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. Patent No. 4,879,231; and Sreekrishna et al. (1987) Gene 59:115, the entire contents of which are incorporated herein by reference in their entirety). Transfection of animal cells can be characterized, for example, by using calcium phosphate, electroporation, heat shock, liposomes, or transfection reagents such as or to introduce the vector into the cell, or by contacting the naked nucleic acid vector with the cell in solution to introduce the vector into the cell (see, for example, Sambrook et al., supra).

[0285] Expression systems that can be used for the small-scale or large-scale production of the peptides (or fusion proteins) described herein include, but are not limited to, microorganisms such as bacteria (e.g., Escherichia coli (E. coli) and Bacillus subtilis) transformed with recombinant phage DNA, plasmid DNA, or cosmid DNA expression vectors; fungi (e.g., yeast (e.g., Saccharomyces and Pichia)) transformed with recombinant yeast expression vectors; insect cell systems infected with recombinant viral expression vectors (e.g., baculovirus); plant cell systems infected with recombinant viral expression vectors (e.g., cauliflower mosaic virus (CaMV) and tobacco mosaic virus (TMV)) or with recombinant plasmid expression vectors (e.g., Ti plasmid); or mammalian cell systems containing recombinant expression constructs comprising promoters derived from mammalian cells (e.g., the metallothionein promoter) or from mammalian viruses (e.g., the adenovirus late promoter, the CMV promoter, the SV40 promoter, or the vaccinia virus 7.5K promoter) (e.g., COS, CHO, BHK, 293, VERO, HeLa, MDCK, WI38, and NIH 3T3 cells). Also used as host cells are primary or secondary cells directly obtained from mammals, which are transfected with plasmid vectors or infected with viral vectors (e.g., viral vectors such as herpesvirus, retrovirus, vaccinia virus, attenuated vaccinia virus, canarypox virus, adenovirus, and adenovirus-associated virus, etc.).

[0286] As described above, after any expression of the peptides (or fusion proteins) described herein, the peptides (or fusion proteins) can be isolated from the cultured cells or from the culture medium of the cultured cells using standard techniques (see, Sambrook et al., supra). Methods for isolating proteins are known in the art and include, for example, liquid chromatography (e.g., HPLC), affinity chromatography (e.g., metal chelate or immunoaffinity chromatography), ion exchange chromatography, hydrophobic interaction chromatography, precipitation or differential solubilization.

[0287] Smaller peptides (e.g., peptides having fewer than 200 (e.g., fewer than 175, fewer than 150, fewer than 125, fewer than 100, fewer than 90, fewer than 80, fewer than 70 or fewer than 60) amino acids) can be synthesized by standard chemical methods such as FMOC solid phase synthesis chemistry (see Example 1).

[0288] The peptides (and fusion proteins) described herein can be isolated (but not necessarily). The term "isolated", when applied to any of the peptides (or fusion proteins) described herein, refers to a peptide and its fragments that have been separated or purified from its natural accompanying components (e.g., proteins or other naturally occurring biomolecules or organic molecules), (or for use in a composition, macromolecular complex). It is understood that recombinant molecules (e.g., recombinant peptides) will generally be "isolated". Generally, a peptide (or fragment or macromolecular complex) is isolated when the peptide comprises at least 60% by weight of the total molecules of the same type in the 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 comprises at least 60% by weight of the total proteins in a preparation or sample. In some embodiments, the molecules in the preparation consist of at least 75%, at least 90, or at least 99% by weight of the total molecules of the same type in the preparation.

[0289] Similarly, a peptide coding sequence, a fusion protein coding sequence, or a vector containing the sequences described herein can be isolated. When applied to any of the peptide coding sequences, fusion protein coding sequences, or vectors described herein, the term "isolated" refers to a peptide coding sequence, a fusion protein coding sequence, or a vector, or a fragment thereof, that has been separated or purified from the components that naturally accompany it (e.g., nucleic acids, proteins, or other naturally occurring biomolecules or organic molecules). It is understood that recombinant molecules (e.g., recombinant vectors or peptide coding sequences or fusion protein coding sequences) will generally be "isolated". Typically, a peptide coding sequence, a fusion protein coding sequence, or a vector (or a 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 a vector described herein is considered isolated when it constitutes at least 60% by weight of the total nucleic acids in a preparation or sample. In some embodiments, the molecules in a preparation consist of at least 75%, at least 90%, or at least 99% by weight of the total molecules of the same type in the preparation.

[0290] In some embodiments, an isolated peptide, fusion protein, peptide coding sequence, fusion protein coding sequence, or vector can be frozen, lyophilized, or immobilized and stored under suitable conditions that allow the molecule to remain active (e.g., the ability of the peptide to bind to MHC molecules such as MHC class I molecules, or the ability of the vector to support the expression of the peptide in a cell).

[0291] Additional Processing of Peptides

[0292] After the expression or synthesis of any of the peptides (or fusion proteins) described herein, the peptide (or fusion protein) can be further processed. Additional processing can include chemical or enzymatic modification of the peptide (or fusion protein), or, in the case of a modified peptide (or fusion protein), the processing can include enzymatic or chemical alteration of the existing modification, or both. Additional processing of the peptide can include the addition (covalent or non-covalent) of heterologous amino acid sequences, such as, but not limited to, any of the heterologous amino acid sequences described above. Enzymatic treatment can involve contacting the peptide with, for example, one or more proteases, phosphatases, or kinases under conditions that allow modification of the peptide. Enzymatic treatment can involve contacting the peptide with one or more enzymes capable of glycosylating the peptide or modifying the glycosylation of the peptide (e.g., oligosaccharyltransferase or mannosidase).

[0293] The processing may include, for example, adding a detectable tag to the peptide. For example, the peptide can be detectably labeled with: an enzyme (e.g., horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase), a fluorescent substance (e.g., umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine, fluorescein, dansyl chloride, allophycocyanin (APC), or phycoerythrin), a luminescent substance (e.g., lanthanide elements or their chelates), a bioluminescent substance (e.g., luciferase, luciferin, or aequorin), or a radionuclide (e.g., 3 H, 32 P, 33 P, 125 I or 35 S).

[0294] The processing may also involve coupling the peptide (or fusion protein) to a polymer (e.g., a poly(alkylene) glycol moiety such as a polyethylene glycol moiety). In some embodiments, the polymer can be coupled to the peptide at a site that is the N-terminus on the peptide. In some embodiments, the peptide can contain one or more internal amino acid insertions that provide internal polymer conjugation sites to which the polymer can be conjugated.

[0295] Pharmaceutical Compositions

[0296] The peptides, fusion proteins, and nucleic acids encoding the peptides or fusion proteins described herein can be incorporated into pharmaceutical compositions. The compositions generally include one or more peptides (and / or nucleic acids encoding the peptides) or a pharmaceutically acceptable carrier. As used herein, the phrase “pharmaceutically acceptable carrier” includes solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and absorption delaying agents, and the like that are compatible with pharmaceutical administration. One or more peptides can be formulated as syrups, elixirs, suspensions, powders, granules, tablets, capsules, lozenges, troches, aqueous solutions, creams, ointments, lotions, gels, emulsions, etc. Supplementary active compounds (e.g., one or more chemotherapeutic agents) can also be included in the compositions. Preferably, the compositions contain two or more (e.g., 2, 3, 4, 5, or 6) of those described herein. The compositions can also include immunogenic peptides other than one peptide disclosed herein, e.g., peptides from WT1 or derivatives thereof, e.g., as described herein. Other immunogenic peptides include, but are not limited to, immunogenic peptides from MUC1, immunogenic peptides from gp100, immunogenic peptides from TRP-2, immunogenic peptides from MAG1, immunogenic peptides from NY-ESO1, immunogenic peptides from HER-2; and immunogenic peptides from AIM2.

[0297] A pharmaceutical composition is generally formulated to be compatible with its intended route of administration. Routes of administration include, for example, oral, rectal, and parenteral, such as intravenous, intramuscular, intradermal, subcutaneous, inhalation, transdermal, or transmucosal. The excipients for a parenteral suspension may include the following components: sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol, or other synthetic solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium metabisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetic, citric, or phosphoric acid, and agents for adjusting tonicity such as sodium chloride or dextrose. The pH may be adjusted with an acid or a base, such as hydrochloric acid or sodium hydroxide. The composition may be enclosed in an ampoule, disposable syringe, or multi-dose vial made of glass or plastic.

[0298] 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 dispersions. 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 must be a fluid to the extent that easy syringability is obtained. It must be stable under the conditions of preparation and storage and must maintain its activity against microbial contamination such as bacteria and fungi. The carrier can be a solvent or a dispersion medium containing, for example, water, ethanol, polyols (such as glycerin, propylene glycol, and liquid polyethylene glycol and the like), and suitable mixtures thereof. The proper fluidity can be maintained by the use of coatings such as lecithin, by maintaining the desired particle size in the case of a dispersion, and by the use of surfactants. Prevention of microbial contamination can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it is desirable to include isotonic agents in the composition, for example, sugars, polyols, such as mannitol, sorbitol, sodium chloride. Prolonged absorption of injectable compositions can be achieved by including agents that prolong absorption, for example, aluminum monostearate and gelatin.

[0299] If desired, after filtration sterilization, a sterile injectable solution can be prepared by incorporating the required amount of one or more peptides (or one or more nucleic acids encoding the peptides) with one of the ingredients or a combination of ingredients listed above into a suitable solvent. Generally, it is prepared by incorporating the peptide (or fusion protein or nucleic acid encoding the peptide) into a sterile vehicle that comprises a basic dispersion medium and the required other ingredients from those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation can include vacuum drying or lyophilization, which yields a powder of the active ingredient and any additional required ingredients from a previously sterile filtered solution.

[0300] Oral compositions generally include an inert diluent or an edible carrier. For purposes of oral therapeutic administration, one or more peptides (or fusion proteins) can be combined 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 as a mouthwash. Pharmaceutically compatible binding agents and / or adjuvant substances can be included as part of the composition. Tablets, pills, capsules, troches, and the like can contain any of the following ingredients, or compounds of a similar nature: binders such as microcrystalline cellulose, gum tragacanth, or gelatin; excipients such as starch or lactose, disintegrating agents such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate or sterotes; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; flavoring agents such as peppermint, methyl salicylate, or orange flavor.

[0301] Powders and tablets can contain from 1% to 95% (w / w) of the individual peptide or a mixture of two or more peptides. In certain embodiments, the peptides can range from about 5% to 70% (w / w). Suitable carriers are magnesium carbonate, magnesium stearate, marble, sugar, galactose, gelatin, dextrin, starch, gelatin, gum tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, and the like. The term "article" is intended to include the formulation of a peptide (or nucleic acid) using an encapsulating substance as a carrier, which provides a capsule in which the peptide, with or without other carriers, is surrounded by the carrier, such that the peptide is bound to the carrier. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0302] If desired, an aqueous solution for oral administration can be prepared by dissolving the active ingredient in water and adding suitable coloring agents, flavoring agents, stabilizers, and thickening agents. An aqueous suspension for oral administration can be prepared by dispersing the finely divided active component in water containing a viscous substance such as natural or synthetic rubber, resin, carboxymethylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents.

[0303] For administration by inhalation, the peptide may be delivered in the form of an aerosol spray from a pressurized container, dispenser, or nebulizer containing a suitable propellant, e.g., a gas such as carbon dioxide.

[0304] Systemic administration can also be effected 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, detergents, bile salts, and fusidic acid derivatives for transmucosal administration. Transmucosal administration can be effected using a nasal spray or a suppository. For transdermal administration, the peptide (or fusion protein or nucleic acid) can be formulated as an ointment, salve, gel, or cream, which are generally known in the art.

[0305] The peptide can also be prepared in the form of a suppository (e.g., containing conventional suppository bases such as cocoa butter and other glycerides) or a retention enema for rectal delivery.

[0306] In one embodiment, the peptide (or fusion protein or nucleic acid) can be formulated with a carrier that will protect the peptide (fusion protein or nucleic acid) from rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene-vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. The methods for preparing such formulations will be apparent to those skilled in the art. The materials are also commercially available from Alza Corporation and Nova Pharmaceuticals, Inc. Liposome suspensions (including targeted liposomes, e.g., liposomes containing monoclonal antibodies specific for an APC antigen to which the APC is specific) can be used as pharmaceutically acceptable carriers. These can be prepared by methods known to those skilled in the art, e.g., as described in U.S. Patent No. 4,522,811.

[0307] It may be advantageous to formulate oral or parenteral compositions in unit dosage form for ease of administration and uniformity of dosage. As used herein, a unit dosage form refers to a physically discrete unit suitable as a single dose for the subject to be treated; each unit containing a predetermined quantity of the peptide (or fusion protein or nucleic acid) calculated to produce the desired therapeutic effect, in association with the required pharmaceutical carrier. The unit dosage forms can also be accompanied by directions for use.

[0308] Nucleic acid molecules encoding the peptide (or fusion protein) can be inserted into a vector and used as a gene therapy vector (as described above). The gene therapy vector can be delivered to a subject by, for example, intravenous injection, local administration, or stereotactic injection (see, e.g., Chen, et al. (1994) Proc. Natl. Acad. Sci. USA 91:3054-3057). Pharmaceutical compositions of the gene therapy vector can include the gene therapy vector in an acceptable diluent, or can include a slow release matrix in which the gene delivery vehicle is embedded. Optionally, where the complete gene delivery vector can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical composition can include one or more cells that produce the gene delivery system (see under "ex vivo methods").

[0309] Additional examples of gene delivery vehicles include, but are not limited to, liposomes, biocompatible polymers, including natural and synthetic polymers; lipoproteins; polypeptides; polysaccharides; lipopolysaccharides; artificial virus envelopes; metal particles; bacteria; viruses such as baculoviruses, adenoviruses, and retroviruses; antibiotics; cosmids; plasmids; fungal vectors; and other recombinant vectors commonly used in the art, which have been described for expression in a variety of eukaryotic and prokaryotic hosts, and can be used for gene therapy as well as for simple protein expression.

[0310] Examples of viral vectors include retroviral vectors, adenoviral vectors, adeno-associated viral vectors, alphavirus vectors, and the like. Liposomes containing targeting moieties such as antibodies or fragments thereof can also be used to prepare pharmaceutical compositions for delivering nucleic acids to a subject.

[0311] Any of the pharmaceutical compositions described herein and the described methods of administration described below can be included in a container, pack, or dispenser.

[0312] MHC Molecule Multimer Compositions and Methods of Using the Compositions

[0313] The present disclosure is also characterized by compositions comprising: (i) one or more of any of the peptides described above, and (ii) a major histocompatibility complex (MHC) molecule multimer. The multimer comprises two or more (e.g., three, four, five, six, seven, eight, nine, or 10 or more) intact MHC molecules or peptide-binding regions of MHC molecules. One or more peptides can be associated with (e.g., covalently or non-covalently bound to) the MHC molecule multimer.

[0314] The MHC molecules of the multimer can be class I MHC molecules (e.g., HLA-A molecules such as HLA-A2 or HLA-A24 molecules) or MHC class II molecules. The MHC molecules can be mammalian (e.g., rodent, non-human primate, human or any other mammal described herein) MHC molecules.

[0315] Two or more MHC molecules (or the peptide-binding regions of MHC molecules) in the multimer can be from the same MHC molecule or from different MHC molecules. For example, an MHC molecule multimer can comprise five MHC molecules, three of which are the same MHC molecule, 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.

[0316] In some embodiments, the above compositions can comprise 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 compositions can also include immunogenic peptides other than the one disclosed herein, e.g., peptides from WT1 or derivatives thereof. Other immunogenic peptides include, but are not limited to, immunogenic peptides from MUC1, immunogenic peptides from gp100, immunogenic peptides from TRP-2, immunogenic peptides from MAG1, immunogenic peptides from NY-ESO1, immunogenic peptides from HER-2; and immunogenic peptides from AIM2.

[0317] The compositions can also be conjugated 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 substances, luminescent substances, bioluminescent substances or radionuclides) and methods for linking the detectable label to the peptides or the MHC molecules described above.

[0318] MHC multimer compositions can be generated using the peptides described above, as follows: refolding the peptide bound to the HLA molecule in the presence of the corresponding HLA heavy chain and β 2 -microglobulin to generate a trimolecular complex. The complex is then biotinylated at the carboxyl terminus of the heavy chain at a site previously engineered into the heavy chain. Multimer formation is then induced by addition of streptavidin.

[0319] Because T cell receptors are able to recognize specific peptide-MHC complexes among a variety of other peptide-MHC complexes, the MHC multimer compositions described herein can be used, for example, to detect antigen-specific T cells in a population of unrelated T cells (see below). For such assays, the multimers will typically be detectably labeled (see above).

[0320] For example, MHC molecule / peptide complexes of multimers can be used in assays of peripheral blood mononuclear cells for the assessment of antigen-specific CTLs after exposure to an immunogen. MHC multimer complexes can be used to directly visualize antigen-specific CTLs (see, e.g., Ogg et al., Science 279:2103-2106, 1998; and Altman et al., Science 174:94-96, 1996) and to determine the frequency of antigen-specific CTL populations in a sample of peripheral blood mononuclear cells. In one example, streptavidin with a detectable label used to multimerize the MHC multimer can be used to label T cells that bind to the MHC molecule / peptide complex of the multimer. To this end, cells treated with the multimer are exposed to, e.g., a tag (e.g., a fluorophore conjugated to biotin). The cells can then be readily separated or assayed, e.g., using a flow cytometer.

[0321] Applications

[0322] The peptides, fusion proteins (and their pharmaceutical compositions), MHC multimers containing the compositions, kits, and articles described herein can be used in a variety of methods. For example, the peptides described herein can be used for: (i) inducing an immune response in a subject (e.g., a subject suffering from cancer); (ii) activating T cells in culture (e.g., central memory T cells and / or effector memory T cells); and / or (iii) treating or even preventing cancer. Cancers include, for example, lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, blood cell cancers, e.g., plasma cell cancers such as multiple myeloma and white blood cells such as AML or CML. The peptides described herein can be used to treat pre-cancerous conditions such as smoldering multiple myeloma. As described above, MHC multimers containing the compositions can be used, e.g., to detect antigen-specific T cells in an unrelated T cell population.

[0323] Although the peptides (or their pharmaceutical compositions), MHC multimers containing the compositions, kits, or articles are not intended to be limited to any particular embodiment described herein, exemplary methods in which these reagents can be used are provided below.

[0324] Methods for Inducing an Immune Response

[0325] The present disclosure features a variety of methods for inducing an immune response in a subject. Methods for inducing an immune response in a subject can include the step of administering to the subject one or more of any of the peptides described herein or any of the pharmaceutical compositions described herein. The immune response can be a CD8 + T cell, CD4 +T cells, cytotoxic T lymphocytes (CTLs), T H 1 responses, T H 2 responses, or a combination of the two types of responses.

[0326] Any of the above methods may also include, for example, methods for treating or preventing (defending against) cancer in a subject (e.g., plasma cell disorders such as multiple myeloma or Waldenstrom macroglobulinemia or any other cancer expressing XBP1, CD138, or CS1). When the terms “prevent” as used herein are related to a given treatment for a given disorder, they mean that the treated subject will not develop the disorder at a clinically observable level (e.g., the subject will not exhibit one or more symptoms of the disorder, or in the case of cancer, the subject will not develop a detectable level of cancer).

[0327] As used herein, the term “treat / therapy” in reference to a subject afflicted with a disorder (e.g., cancer) is used in connection with a given treatment for the given disorder, wherein at least one symptom of the disorder is cured, healed, alleviated, slowed, altered, corrected, improved, or ameliorated. Treatment includes administering an amount of a composition effective to alleviate, slow, alter, correct, improve, or affect the disorder or a symptom of the disorder. Treatment may inhibit the worsening or deterioration of the symptoms of the disorder in the subject, or may cause the disorder to progress more slowly and / or to a lesser extent compared to treatment in the absence thereof (e.g., fewer symptoms or a lower number of cancer cells in the subject). For example, if treatment is given during the course of a disorder, e.g., during early diagnosis of cancer where the development of established manifestations of the disorder (advanced cancer) is anticipated (e.g., detection of a small number of cancer cells in a sample from the subject), and results in the subject experiencing fewer and / or less severe symptoms of the disorder than would be expected, the treatment will be considered to have “treated” the disorder. When a subject exhibits only mild and apparent symptoms of cancer, treatment may “treat” the cancer (e.g., plasma cell disorders such as multiple myeloma or Waldenstrom macroglobulinemia).

[0328] In one embodiment, the cancer is a cancer as described herein. For example, the cancer can be bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma, and large cell carcinoma)), genitourinary cancer, such as ovarian cancer (including fallopian tube cancer, endometrial cancer, and peritoneal cancer), cervical cancer, prostate cancer, and testicular cancer, lymphatic system cancer, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer, or lower gastric cancer), gastrointestinal cancer (e.g., anal cancer or bile duct cancer), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancer (e.g., glioblastoma multiforme), and head and neck cancer (e.g., nasopharyngeal cancer).

[0329] Generally, the peptide to be delivered to the subject can be suspended in a pharmaceutically acceptable carrier (e.g., saline) and administered orally, rectally, or parenterally, e.g., by intravenous injection, subcutaneous injection, intramuscular injection, intrathecal injection, intraperitoneal injection, rectal injection, vaginal injection, intranasal injection, intragastric injection, intratracheal injection, or intralung injection.

[0330] Administration can be by periodic injection of boluses of the pharmaceutical composition, or by intravenous or intraperitoneal administration from an external reservoir (e.g., an IV bag) or an internal reservoir (e.g., a bioerodible implant, a bioartificial trachea, or a population of reagent-producing cells implanted). See, e.g., U.S. Patent Nos. 4,407,957, 5,798,113, and 5,800,828, each incorporated herein by reference in its entirety.

[0331] Generally, the dose of the desired peptide or nucleic acid depends on the choice of administration route; the nature of the formulation; the nature or severity of the subject's disease; the subject's immune status; the size, weight, surface area, age, and sex of the subject; other drugs being administered; and the judgment of the attending physician.

[0332] The suitable dosage range of the peptide for inducing an immune response is from 0.000001 mg to 10 mg per kg of the subject of the reagent or antigenic / immunogenic composition. Wide variations in the required dosage are expected depending on the different kits and the differential efficiencies of the different routes of administration. For example, nasal administration or rectal administration may require higher dosages than administration by intravenous injection. As is well known in the art, the differences in these dosage levels can be adjusted using standard empirical procedures for optimization. The administration can be single or multiple (e.g., 2-fold, 3-fold, 4-fold, 6-fold, 8-fold, 10-fold, 20-fold, 50-fold, 100-fold, 150-fold or higher). For example, the peptide can be administered as an initial immunization and then subsequently administered one or more times as a booster immunization.

[0333] To optimize the efficacy (e.g., the potency of one or more peptides or nucleic acids encoding the peptides to induce an immune response in a subject), the composition comprising the peptide or nucleic acid can be administered for the first time in different dosing regimens. The unit dosage and regimen depend on factors including, for example, the species of the mammal, its immune status, and the body weight of the mammal.

[0334] The frequency of administering the pharmaceutical composition (e.g., the pharmaceutical composition described herein) is within the clinical judgment of the person skilled in the art and medical practitioners (e.g., a doctor or a nurse). Generally, the dosing regimen is established through clinical trials that can establish the optimal dosing parameters. However, the practitioner can alter the dosing regimen based on the age, health, weight, gender, and medical status of the subject.

[0335] In some embodiments, the pharmaceutical composition can be administered to the subject at least two times (e.g., three times, four times, five times, six times, seven times, eight times, nine times, 10 times, 11 times, 12 times, 15 times or 20 times or more). For example, the pharmaceutical composition can be administered to the subject once a month for three months; once a week for one 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.

[0336] In some embodiments, the reagent can be administered together with an immunomodulator such as a Toll receptor ligand or an adjuvant (see below).

[0337] As defined herein, a "therapeutically effective amount" of a peptide or nucleic acid encoding a peptide is an amount of the peptide or nucleic acid that is capable of generating an immune response in a subject being treated. The therapeutically effective amount of a peptide (i.e., the effective dose) includes amounts of the agent in milligrams, micrograms, nanograms, or picograms per kilogram of subject or sample weight (e.g., from about 1 nanogram per kilogram to about 500 micrograms per kilogram, from about 1 microgram per kilogram to about 500 milligrams per kilogram, from about 100 micrograms per kilogram to about 5 milligrams per kilogram, or from about 1 milligram per kilogram to about 50 micrograms per kilogram). The therapeutically effective amount of a nucleic acid also includes amounts of the agent in micrograms, nanograms, or picograms per kilogram of subject or sample weight (e.g., from about 1 nanogram per kilogram to about 500 micrograms per kilogram, from about 1 microgram per kilogram to about 500 micrograms per kilogram, from about 100 micrograms per kilogram to about 500 micrograms per kilogram, or from about 1 microgram per kilogram to about 50 micrograms per kilogram).

[0338] 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 generating an immune response in a subject being treated against cancer cells (e.g., breast cancer cells, colon cancer cells, pancreatic cancer cells, prostate cancer cells, multiple myeloma cells, or Waldenstrom macroglobulinemia), said immune response being capable of preventing the development of cancer in the subject, or being capable of substantially reducing the chance that the subject will develop or continue to develop cancer (see above). The prophylactically effective amount of a peptide (i.e., the effective dose) includes amounts of the agent in milligrams, micrograms, nanograms, or picograms per kilogram of subject or sample weight (e.g., from about 1 nanogram per kilogram to about 500 micrograms per kilogram, from about 1 microgram per kilogram to about 500 milligrams per kilogram, from about 100 micrograms per kilogram to about 5 milligrams per kilogram, or from about 1 milligram per kilogram to about 50 micrograms per kilogram). The prophylactically effective amount of a nucleic acid also includes amounts of the agent in micrograms, nanograms, or picograms per kilogram of subject or sample weight (e.g., from about 1 nanogram per kilogram to about 500 micrograms per kilogram, from about 1 microgram per kilogram to about 500 milligrams per kilogram, from about 100 micrograms per kilogram to about 5 milligrams per kilogram, or from about 1 milligram per kilogram to about 50 micrograms per kilogram).

[0339] The subject can be any animal capable of mounting 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., a chimpanzee, baboon or monkey), mouse, rat, rabbit, guinea pig, gerbil, hamster, horse, domestic animal type (e.g., cow, pig, sheep or goat), dog, cat or whale. The subject can be one suffering from cancer, suspected of having cancer or at risk of developing cancer, such as multiple myeloma, Waldenstrom macroglobulinemia, or any type of cancer expressing XBP1, CD138 or CS-1 (e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric 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 cancer, such as breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, e.g., AML or CML, multiple myeloma or Waldenstrom macroglobulinemia.

[0340] The method can further comprise the step of determining, in a subject to whom one or more peptides (or nucleic acids) are administered, that one or more cancer cells of the subject's cancer (e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia or plasma cell disorders such as multiple myeloma or Waldenstrom macroglobulinemia) express XBP1, CD138 or CS-1. Expression of these proteins includes mRNA and protein expression. Methods for detecting protein and mRNA expression in cells are known in the art and include, for example, enzyme-linked immunosorbent assay (ELISA), Western blotting techniques or immunohistochemical techniques for detecting proteins and reverse transcriptase polymerase chain reaction (RT-PCR) or northern blotting techniques for detecting mRNA. (See, Sambrook et al., supra).

[0341] A peptide or composition can be used in combination with other known therapies. As used herein, "combination" administration means delivering two (or more) different treatments to a subject during the course of a disorder in the subject, e.g., delivering two or more treatments after diagnosing the subject with a disorder and before curing or eliminating the disorder or discontinuing treatment delivery for other reasons. In some embodiments, when delivery of the second therapy is initiated, delivery of one therapy is still occurring, such that there is an overlap in administration. This is sometimes referred to as "simultaneous" or "concurrent" delivery. In further embodiments, delivery of one therapy ends before delivery of the other therapy begins. In some embodiments in either case, the combination of therapies is more effective. For example, the second therapy is more effective, e.g., an equivalent effect is observed with less of the second therapy as compared to what would be observed when administering the second therapy without the first therapy, or the second therapy reduces symptoms to a greater extent. In some embodiments, the delivery is such that the reduction in symptoms or other parameters associated with the disorder is higher as compared to what is observed with delivery of one therapy without the other. The effects of the two therapies can be partially additive, fully additive, or more than additive. The delivery can be such that the effect of the first therapy being delivered is still detectable when the second therapy is delivered.

[0342] The additional therapy can be, for example, surgery, one or more chemotherapeutic agents, one or more forms of ionizing radiation, and / or one or more immunomodulators.

[0343] One or more forms of ionizing radiation can be γ-irradiation, X-irradiation, or β-irradiation.

[0344] Exemplary classes of chemotherapeutic agents include, for example, those described below:

[0345] Alkylating agents (including, but not limited to, nitrogen mustards, ethyleneimine derivatives, alkyl sulfonates, nitrosoureas, and triazines): nitrogen mustard (Aminouracil Uracil nitrogen )、nitrogen mustard (chlormethine) cyclophosphamide ( Revimmune TM )、ifosfamide melphalan chlorambucil pipobroman triethylenemelamine triethylenethiophosphoramide, temozolomide thiotepa busulfan Carmustine .Lomustine Streptozocin and Dacarbazine

[0346] Anti-EGFR antibodies (e.g., Cetuximab and Panitumumab

[0347] Anti-HER-2 antibodies (e.g., Trastuzumab

[0348] Antimetabolites (including, but not limited to, folic acid antagonists (also referred to herein as antifolates), pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors: Methotrexate 5-Fluorouracil Floxuridine Cytarabine( Tarabine PFS), 6-Mercaptopurine 6-Mercaptopurine (Thioguanine ), Fludarabine Phosphate Pentostatin Pemetrexed Raltitrexed Cladribine Clofarabine Mercaptopurine Capecitabine Nelarabine Azacitidine and Gemcitabine Preferred antimetabolites include, for example, 5-fluorouracil Floxuridine Capecitabine Pemetrexed Raltitrexed and Gemcitabine

[0349] Vinca alkaloids: Vinblastine Vincristine Vindesine Vinorelbine

[0350] Platinum-based agents: Carboplatin Cisplatin Oxaliplatin

[0351] Anthracyclines: Daunomycin Doxorubicin Epirubicin Idarubicin . Mitoxantrone Valrubicin Preferred anthracyclines include daunomycin and doxorubicin

[0352] Topoisomerase inhibitors: Topotecan Irinotecan Etoposide Teniposide Lamellarin D, SN-38, camptothecin.

[0353] Taxanes: Paclitaxel Docetaxel Larotaxel, cabazitaxel.

[0354] Epothilones: Ixabepilone, epothilone B, epothilone D, BMS310705, dehydelone, ZK-Epothilone (ZK-EPO).

[0355] 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).

[0356] Antibiotics: Actinomycin Bleomycin Hydroxyurea Mitomycin

[0357] Immunomodulators: Lenalidomide Thalidomide

[0358] Immune cell antibodies: Alemtuzamab Gemtuzumab Rituximab Tositumomab

[0359] Interferons (e.g., IFN-α or

[0360] Interleukins: IL-1, IL-24, IL-12.

[0361] HSP90 inhibitors (e.g., geldanamycin or any derivatives thereof). In certain embodiments, the HSP90 inhibitor is selected from geldanamycin, 17-alkylamino-17-demethoxygeldanamycin (“17-AAG”), or 17-(2-dimethylaminoethyl)amino-17-demethoxygeldanamycin (“17-DMAG”).

[0362] Angiogenesis inhibitors, including but not limited to 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 O2) (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-MyersSquibb), 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 Pharmaceuticals), 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 (ImCloneLLC), 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 (AlconResearch), 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.), VEGFTrap (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.

[0363] Anti-androgens, including but not limited to nilutamide and bicalutamide

[0364] Anti-estrogens, including but not limited to tamoxifen toremifene letrozole testolactone anastrozole bicalutamide exemestane flutamide fulvestrant raloxifene and raloxifene hydrochloride.

[0365] Hypercalcemia agents, including but not limited to, gallium(III) nitrate hydrate and disodium pamidronate

[0366] Apoptosis inducers, including but not limited to, ethanol, 2-[[3-(2,3-dichlorophenoxy)propyl]amino]-(9Cl), gambogic acid, embelin, and arsenic trioxide

[0367] Aurora kinase inhibitors, including but not limited to, binucleine 2.

[0368] Bruton tyrosine kinase inhibitors, including but not limited to aspergillic acid.

[0369] Calcineurin inhibitors, including but not limited to, cypermethrin, deltamethrin, fenvalerate, and tyrosine phosphorylation inhibitor 8.

[0370] CaM kinase II inhibitors, including but not limited to, 5-isoquinolinesulfonic acid, 4-[{2S)-2-[(5-isoquinolinylsulfonyl)methylamino]-3-oxo-3-{4-phenyl-1-piperazinyl)propyl]phenyl ester, and benzenesulfonamide.

[0371] CD45 tyrosine phosphatase inhibitors, including but not limited to phosphonic acid.

[0372] CDC25 phosphatase inhibitors, including but not limited to, 1,4-naphthoquinone, 2,3-bis[(2-hydroxyethyl)thio]-(9Cl).

[0373] CHK kinase inhibitors, including but not limited to debromohymenialdisine.

[0374] Cyclooxygenase inhibitors, including but not limited to 1H-indole-3-acetamide, 1-(4-chlorobenzoyl)-5-methoxy-2-methyl-N-(2-phenylethyl)-(9Cl), 5-alkyl-substituted 2-aryl aminophenylacetic acid and its derivatives (e.g., celecoxib rofecoxib etoricoxib lumiracoxib valdecoxib or 5-alkyl-2-aryl aminophenylacetic acid).

[0375] cRAF kinase inhibitors, including but not limited to 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).

[0376] Cyclin-dependent inhibitors, including but not limited to olomoucine and its derivatives, purvalanol B, roscovitine Isatin, kenpaullone, purvalanol A, and isatin-3'-oxime.

[0377] Cysteine protease inhibitors, including but not limited to 4-morpholinecarboxamide, N-[(1S)-3-fluoro-2-oxo-1-(2-phenylethyl)propyl]amino]-2-oxo-1-(benzyl)ethyl]-(9Cl).

[0378] DNA intercalating agents, including but not limited to mithramycin and daptomycin

[0379] DNA strand breakers, including but not limited to bleomycin

[0380] E3 ligase inhibitors, including but not limited to N-((3,3,3-trifluoro-2-trifluoromethyl)propanoyl)sulfonamide.

[0381] EGF pathway inhibitors, including but not limited to tyrosine phosphorylation inhibitor 46, EKB-569, erlotinib gefitinib lapatinib and those compounds disclosed generally and specifically in the following: 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, US 5,747,498, WO 98 / 10767, WO 97 / 30034, WO 97 / 49688, WO 97 / 38983, and WO 96 / 33980.

[0382] Farnesyl transferase inhibitors, including but not limited to A-hydroxyfarnesylphosphonic acid, butyric acid, 2-[(2S)-2-[[(2S,3S)-2-[[(2R)-2-amino-3-mercaptopropyl]amino]-3-methylphenyl]oxy]-1-oxo-3-phenylpropyl]amino]-4-(methylsulfonyl)-1-methylethyl ester (2S)-(9Cl), and manumycin A.

[0383] Flk-1 kinase inhibitors, including but not limited to 2-acrylamide, 2-cyano-3-[4-hydroxy-3,5-bis(1-methylethyl)phenyl]-N-(3-phenylpropyl)-(2E)-(9Cl).

[0384] Glycogen synthase kinase-3 (GSK3) inhibitors, including but not limited to indirubin-3'-oxime.

[0385] Heat shock protein 90 (Hsp90) chaperone modulators, including but not limited to AUY922, STA-9090, ATI13387, MCP-3100, IPI-504, IPI-493, SNX-5422, Debio0932, HSP990, DS-2248, PU-H71, 17-DMAG (Alvespimycin), and XL888.

[0386] Histone deacetylase (HDAC) inhibitors, including but not limited to hydroxamic acid (SAHA), [4-(2-aminobenzoyl)-benzyl]-carbamic acid pyrimidin-3-ylmethyl ester and its derivatives, butyric acid, pyroxamide, trichostatin A, oxamflatin, apicidin, desacetylcephalosporin C, depudecin, trapoxin, and the compounds disclosed in WO 02 / 22577.

[0387] I-kappa B-alpha kinase inhibitors (IKK), including but not limited to 2-acrylonitrile, 3-[(4-tolyl)sulfonyl]-(2E)-(9Cl).

[0388] Imidazotetrazinones, including but not limited to temozolomide ( and its derivatives (e.g., as generally and specifically disclosed in US 5,260,291) and mitozolomide.

[0389] Insulin-like growth factor pathway inhibitors, such as IGF inhibitors or IGF receptor (IGFR1 or IGFR2) inhibitors, including but not limited to, 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).

[0390] Insulin tyrosine kinase inhibitors, including but not limited to hydroxy-2-naphthalenemethylphosphonic acid.

[0391] c-Jun-N-terminal kinase (JNK) inhibitors, including but not limited to pyrazoloanthrone and epicatechin gallate.

[0392] Mitogen-activated protein kinase (MAP) inhibitors, including but not limited to benzenesulfonamide, N-[2-[[[3-(4-chlorophenyl)-2-propenyl]methyl]amino]methyl]phenyl]-N-(2-hydroxyethyl)-4-methoxy-(9Cl).

[0393] MDM2 inhibitors, including but not limited to trans-4-iodo-4'-boronyl-chalcone.

[0394] MEK inhibitors, including but not limited to succinonitrile, bis[amino[(2-aminophenyl)thio]methylene]-(9Cl).

[0395] MMP inhibitors, including but not limited to actinomycin, epicatechin gallate, collagen peptide mimetic and non-peptide mimetic inhibitors, tetracycline derivative marimastat prinomastat, incyclinide shark cartilage extract AE-941 Tanomastat, TAA211, MMI270B or AAJ996.

[0396] mTor inhibitors, including but not limited to rapamycin and its analogs and derivatives, AP23573 (also known as ridaforolimus, deforolimus or MK-8669), CCI-779 (also known as temsirolimus) and SDZ-RAD.

[0397] NGFR tyrosine kinase inhibitors, including but not limited to tyrosine phosphorylation inhibitor AG 879.

[0398] p38 MAP kinase inhibitors, including but not limited to phenol, 4-[4-(4-fluorophenyl)-5-(4-pyridyl)-1H-imidazol-2-yl]-(9Cl) and benzamide, 3-(dimethylamino)-N-[3-[(4-hydroxybenzoyl)amino]-4-tolyl]-(9Cl).

[0399] p56 tyrosine kinase inhibitors, including but not limited to daphnetin and tyrosine phosphorylation inhibitor 46.

[0400] PDGF pathway inhibitors, including but not limited to tyrosine phosphorylation inhibitor AG 1296, tyrosine phosphorylation inhibitor 9, 1,3-butadiene-1,1,3-tricarbonitrile, 2-amino-4-(1H-indol-5-yl)-(9Cl), imatinib and gefitinib and those compounds generally and specifically disclosed in European Patent No. 0 564 409 and PCT Publication No. WO 99 / 03854.

[0401] Phosphatidylinositol 3-kinase inhibitors, including but not limited to wortmannin and quercetin dihydrate.

[0402] Phosphatase inhibitors, including but not limited to cantharidic acid, cantharidin and L-leucinamide.

[0403] PKC inhibitors, including but not limited to 1-H-pyrrole-2,5-dione, 3-[1-[3-(dimethylamino)propyl]-1H-indol-3-yl]-4-(1H-indol-3-yl)-(9Cl), bisindolylmaleimide IX, sphingosine, staurosporine and hypericin.

[0404] PKCδ kinase inhibitors, including but not limited to catalpol. Polyamine synthesis inhibitors, including but not limited to DMFO.

[0405] Proteasome inhibitors, including but not limited to aclacinomycin A, gliotoxin and bortezomib

[0406] Protein phosphatase inhibitors, including but not limited to cantharidic acid, cantharidin L-p-bromotetramisole oxalate, 2(5H)-furanone, 4-hydroxy-5-(hydroxymethyl)-3-(1-oxohexadecyl)-(5R)-(9Cl) and benzylphosphonic acid.

[0407] Protein tyrosine kinase inhibitors, including but not limited to tyrosine phosphorylation inhibitor Ag 216, tyrosine phosphorylation inhibitor Ag1288, tyrosine phosphorylation inhibitor Ag 1295, geldanamycin, genistein, and 7H-pyrrolo[2,3-d]pyrimidine derivatives.

[0408] PTP1B inhibitors, including but not limited to L-leucinamide.

[0409] SRC family tyrosine kinase inhibitors, including but not limited to PP1 and PP2.

[0410] Syk tyrosine kinase inhibitors, including but not limited to piceatannol.

[0411] Janus (JAK-2 and / or JAK-3) tyrosine kinase inhibitors, including but not limited to tyrosine phosphorylation inhibitor AG 490 and 2-naphthyl vinyl ketone.

[0412] Retinoids, including but not limited to isotretinoin and tretinoin RNA polymerase II elongation inhibitors, including but not limited to 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole.

[0413] Serine / threonine kinase inhibitors, including but not limited to 2-aminopurine.

[0414] Sterol biosynthesis inhibitors, including but not limited to squalene epoxidase and CYP2D6. VEGF pathway inhibitors, including but not limited to anti-VEGF antibodies, e.g., bevacizumab, and small molecules, e.g., sunitinib Sorafenib ZD6474 (also known as vandetanib) (ZactimaTM), SU6668, CP-547632, AV-951 (tivozanib), and AZD2171 (also known as cediranib) (Recentin TM )

[0415] For example, one or more chemotherapeutic agents can be selected from: cisplatin, carboplatin, procarbazine, mechlorethamine, cyclophosphamide, camptothecin, doxorubicin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosourea, actinomycin D, daunorubicin, doxorubicin, bleomycin, oligomycin, mitomycin, etoposide, verampil, podophyllotoxin, tamoxifen, paclitaxel, thalidomide, lenalidomide, proteasome inhibitors (e.g., bortezomib), HSP90 inhibitors (e.g., tenespinmycin), transplatinum, 5-fluorouracil, vincristine, vinblastine, methotrexate, or analogs of any of the foregoing. Immunomodulators include, for example, various chemokines and cytokines such as interleukin 2 (IL-2), granulocyte / macrophage colony-stimulating factor (GM-CSF), and interleukin 12 (IL-12).

[0416] In one embodiment, the additional therapy is one or more additional immunogenic peptides, for example, one or more immunogenic peptides from WT1 or its derivatives. Exemplary WT1 immunogenic peptides include, but are not limited to, WT1 class 1 epitopes; peptides containing RMFPNAPYL (SEQ ID NO:538) (WT1 126-134) or consisting thereof; peptides containing YMFPNAPYL (SEQ ID NO:539) or consisting thereof; peptides containing RSDELVRHHNMHQRNMTKL (SEQ ID NO:540) (WT1 427-445) or consisting thereof; peptides containing PGCNKRYFKLSHLQMHSRKHTG (SEQ ID NO:541) (WT1 331-352) or consisting thereof; peptides containing SGQARMFPNAPYLPSCLES (SEQ ID NO:542) (WT1 122-140) or consisting thereof; and peptides containing SGQAYMFPNAPYLPSCLES (SEQ ID NO:543) or consisting thereof. Other WT1 immunogenic peptides are described in U.S. Patent No. 7,598,221, the content of which is incorporated herein by reference. Other immunogenic peptides include, but are not limited to, immunogenic peptides from MUC1, immunogenic peptides from gp100, immunogenic peptides from TRP-2, immunogenic peptides from MAG1, immunogenic peptides from NY-ESO1, immunogenic peptides from HER-2; and immunogenic peptides from AIM2.

[0417] The subject may have cancer, be suspected of having cancer, or be at risk of developing cancer, such as lung cancer, liver cancer, bile duct cancer, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia, multiple myeloma, or Waldenstrom macroglobulinemia. A "subject suspected of having cancer" is a subject with one or more symptoms of cancer. The symptoms of cancer are well known to those skilled in the art and generally include, but are not limited to, pain, weight loss, weakness, excessive fatigue, difficulty eating, loss of appetite, chronic cough, severe shortness of breath, coughing up blood, blood in the urine, blood in the stool, nausea, abdominal fullness, abdominal distension, vomiting, ascites, vaginal bleeding, constipation, abdominal distension, colon perforation, acute peritonitis (infection, fever, pain), pain, vomiting blood, profuse sweating, fever, high blood pressure, anemia, diarrhea, jaundice, dizziness, chills, muscle spasms, difficulty swallowing, etc. The symptoms of multiple myeloma specifically include, for example, bone pain (e.g., in the back or ribs), hypercalcemia, excessive thirst or urination, constipation, nausea, loss of appetite, confusion, weakness or numbness in the legs, weight loss, or recurrent infections. The symptoms of Waldenstrom macroglobulinemia include, for example, weakness, enlarged lymph nodes, severe fatigue, nosebleeds, weight loss, and nerve problems.

[0418] As used herein, a subject at risk of developing cancer is a subject who has a susceptibility to cancer, i.e., a genetic susceptibility to developing cancer, such as a mutation in a tumor suppressor gene (e.g., a mutation in BRCA1, p53, RB, or APC), who has been exposed to conditions that can cause cancer or who is currently affected by conditions that can cause cancer. Thus, a subject can also be a "subject at risk of developing cancer" when the subject is exposed to certain compounds at mutagenic or carcinogenic levels (e.g., carcinogenic compounds in cigarette smoke such as acrolein, 4-aminobiphenyl, aromatic amines, benzene, benzo{a}anthracene, benzo{a}pyrene, formaldehyde, hydrazine, polonium-210 (Radon), carbamate, or vinyl chloride). A subject can be "at risk of developing cancer" when the subject is exposed to, for example, high doses of ultraviolet light or X-irradiation, or tumorigenic / associated viruses 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 cancer" when the subject has an inflammation (e.g., chronic inflammation). If, for example, a subject has monoclonal gammopathy of undetermined significance, the subject can be at risk of developing multiple myeloma. A subject can be at risk of developing any of the cancers described herein, e.g., bladder cancer (including accelerated bladder cancer and metastatic bladder cancer), breast cancer (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 cancer (including colorectal cancer), kidney cancer (e.g., renal cell carcinoma (e.g., papillary renal cell carcinoma, clear cell carcinoma, chromophobe cell carcinoma)), liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer (including adenocarcinoma, squamous cell carcinoma, bronchioloalveolar carcinoma, and large cell carcinoma)), genitourinary cancers such as ovarian cancer (including fallopian tube cancer, endometrial cancer, and peritoneal cancer), cervical cancer, prostate cancer, and testicular cancer, lymphatic system cancers, rectal cancer, laryngeal cancer, pancreatic cancer (including exocrine pancreatic cancer), gastric cancer (e.g., gastroesophageal cancer, upper gastric cancer, or lower gastric cancer), gastrointestinal cancers (e.g., anal cancer or cholangiocarcinoma), gallbladder cancer, thyroid cancer, leukemia (e.g., acute myeloid leukemia), neural and glial cell cancers (e.g., glioblastoma multiforme), head and neck cancer, or multiple myeloma.

[0419] Multiple myeloma is a hematological cancer that affects approximately 45,000 people per year in the United States. Multiple myeloma is characterized by multifocal proliferation and clonal expansion of plasma cells in the bone marrow, which can lead to skeletal muscle disorders, serum monoclonal gammopathy, immunosuppression, and end-organ sequelae. Subjects with smoldering multiple myeloma (SMM) are at high risk of developing active multiple myeloma. Although patients with active multiple myeloma are treated with therapies including bone marrow transplantation and chemotherapy, patients with these disease courses generally have a poor prognosis.

[0420] Smoldering multiple myeloma (SMM) is an asymptomatic plasma cell proliferative disorder characterized by monoclonal plasma cell proliferation in the bone marrow and monoclonal protein in the blood and / or urine, without renal disturbances, 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 no 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.

[0421] Currently, there is no active therapy for SMM. Instead, a "watch-and-wait" approach is taken, with treatment starting after progression to a symptomatic disease. For most patients, the following indicate progression: increasing anemia (hemoglobin < 2 g / dL, below the lower limit of normal or < 10 g / dL) and / or skeletal muscle involvement, including bone lesions and / or diffuse osteoporosis.

[0422] Subjects with smoldering multiple myeloma are also subjects at risk of developing multiple myeloma. Smoldering multiple myeloma can be determined by elevated levels of monoclonal protein, such as from the urine or blood of the subject. In addition, subjects with smoldering multiple myeloma may exhibit an increased number of myeloma cells in the bone marrow. Currently, it has been estimated that SMM accounts for approximately 15% of all newly diagnosed MM cases. The median time from diagnosis to progression to symptomatic MM ranges from 2 to 3 years, and the estimated annual risk of progression from SMM to symptomatic MM requiring treatment is 10%. The risk of progression depends on 1) monoclonal protein level ≥ 3 g / dL; 2) BMPC ≥ 10%; and 3) the presence of an abnormal serum free light chain (FLC) ratio. As shown in Table 1, the median time to progression (1.9 years) was significantly shorter in patients who met all three prognostic criteria than in those who met only one of these criteria (10 years) or two of these criteria (5.1 years). Similarly, in those patients who met all 3 prognostic criteria, the proportion of patients who progressed to symptomatic MM within 5 years was significantly higher than in those who met only one or two of these criteria (25% and 51%, respectively).

[0423] Table 4. MM Prognosis

[0424]

[0425] Source: Kyle RA, et al., 2010.

[0426] Recent data have confirmed the importance of other prognostic criteria for SMM, including the presence of an abnormal phenotype of BMPCs, a decrease in one or two of the uninvolved immunoglobulin (Ig) isotypes, and whether the M protein remains stable or deteriorates over time. In the latter case, termed progressive SMM, patients with a progressive increase in serum M protein level have a shorter median time to progression (TTP) compared to those with a stable M protein, 1.3 years versus 3.9 years, respectively.

[0427] In some embodiments, the method may further comprise determining whether an immune response has occurred in a subject after administering to the subject a peptide, nucleic acid, or composition described herein. Suitable methods for determining whether an immune response has occurred in a subject include using immunoassays to detect, for example, the presence of antibodies specific for the peptide in a biological sample from the subject. For example, after administering 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. An immune response can also be detected by determining the presence or amount of activated T cells in the sample. Such assays include, for example, proliferation assays, limiting dilution assays, cytotoxicity assays (e.g., lymphokine release assays or 51 51Cr release assays (as described above).

[0428] In some embodiments, the method may further comprise the step of determining whether the subject has cancer. Suitable methods for such determination depend on the type of cancer to be detected in the subject, but such methods are known in the art. The methods can be qualitative or quantitative. For example, when a subject exhibits two or more symptoms of multiple myeloma (such as any of those described herein), a physician may diagnose the subject with multiple myeloma. The subject having multiple myeloma can also be determined by measuring the serum calcium level, the number of white or red blood cells, or the number of proteins in the urine of the subject.

[0429] Ex vivo methods In vitro strategies for inducing an immune response in a subject can include contacting suitable APCs (e.g., dendritic cells, monocytes, or macrophages) obtained from the subject with a peptide or composition described herein. Optionally, cells can be transfected with a nucleic acid (e.g., an expression vector) encoding one or more peptides and optionally cultured for a period of time and under conditions that permit expression of the peptide. The transfection method will depend on the type of cell and the type of nucleic acid to be transfected into the cell. (See "Nucleic Acids and Methods for Producing Peptides" and Sambrook et al., supra, for the above.) After the contacting or transfection, the cells are returned to the subject.

[0430] The cells can be any broad range of cells that express MHC class I or class II molecules. For example, the cells can include myeloid cells, macrophages, monocytes, dendritic cells, T cells (e.g., T helper cells, CD4 + cells, CD8 + cells or cytotoxic T cells) or B cells.

[0431] Ex vivo methods for stimulating an immune response can include contacting, in vitro, T cells (e.g., from a population of lymphocytes obtained from a subject) with antigen-presenting cells that express MHC molecules that bind to one of the peptides described herein, for a time sufficient to activate the T cells (under conditions sufficient to activate the T cells). After the contacting, the activated T cells are re-introduced into the subject from whom the cells were obtained. Methods for generating APCs that express MHC molecules that bind to one of the peptides described herein are described above in this section.

[0432] In some embodiments of any ex vivo method, the cells can be obtained from the same species (allogeneic) other than the subject, can be contacted with a reagent (or immunogenic / antigenic composition) and administered to the subject.

[0433] Methods for Generating Antibodies in a Subject

[0434] Methods for generating antibodies specific for an immunogen (e.g., one or more of any of the peptides described herein) are known in the art and are described below. For example, antibodies or antibody fragments specific for the peptides 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 the peptides described herein can be used to generate antibodies or antibody fragments.

[0435] Antibodies can be prepared using a peptide by immunizing a suitable subject (e.g., rabbit, goat, mouse, or other mammal such as a human). Suitable immunogenic preparations can comprise, for example, any of the reagents described herein. The preparation can also include adjuvants such as Freund's complete or incomplete adjuvant, alum, RIBI, or similar immunostimulatory agents. Adjuvants also include, for example, cholera toxin (CT), Escherichia coli (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. Immun. 64:974-979), carboxymethylcellulose, polyinosinic-cytidylic acid and poly-L-lysine double-stranded RNA (e.g., poly IC-LC, e.g., hiltonol) combination, water and oil emulsions (e.g., montanide), and proteins (e.g., cytokines, complement, GCSF, GM-CSF). MCT and MLT contain point mutations that substantially attenuate toxicity relative to the parental molecule without substantially impairing adjuvant activity. 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.

[0436] As used herein, the term antibody refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules (i.e., molecules that contain an antigen-binding site that specifically binds to a 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, for example, F(ab) fragments, F(ab') 2 fragments or any other antibody fragments described herein (see below).

[0437] Anti-peptide antibodies can be preparations of monoclonal or polyclonal antibodies. As used herein, the term monoclonal antibody refers to a population of antibody molecules that contain only one antigen-binding site capable of an immune response to a peptide. Thus, a monoclonal antibody composition typically exhibits a single binding affinity for the particular peptide to which it is immunologically responsive.

[0438] Polyclonal anti-peptide antibodies can be prepared by immunizing a suitable subject with a peptide immunogen as described above. Over time, the anti-peptide antibody titers in the immunized subject can be monitored by standard techniques such as enzyme-linked immunosorbent assay (ELISA) using immobilized peptide. If desired, the antibody molecules directed against the peptide can be isolated from a mammal (e.g., from 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 titer is 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, human B cell hybridoma technique (Kozbor et al. (1983) Immunol. Today 4:72) or EBV-hybridoma technique (Cole et al. (1985), Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96). Any of a number of well-known protocols for fusing lymphocytes and an established cell line can be used for the purpose of generating anti-peptide monoclonal antibodies (see, e.g., Current Protocols in Immunology, supra; Galfre et al. (1977) Nature 266:550-52; R.H. Kenneth, in Monoclonal Antibodies: A New Dimension In Biological Analyses, Plenum Publishing Corp., New York, New York (1980); and Lerner (1981) Yale J. Biol. Med., 54:387-402, the disclosures of each of which are incorporated herein by reference in their entirety).

[0439] As an alternative to preparing monoclonal antibody-secreting hybridomas, monoclonal anti-peptide antibodies can be identified and isolated by screening a recombinant combinatorial immunoglobulin library (e.g., an antibody phage display library) described herein with the peptide to isolate immunoglobulin library members that bind the peptide.

[0440] Anti-peptide antibodies (e.g., monoclonal antibodies) can be used to isolate peptides by techniques such as affinity chromatography or immunoprecipitation. Also, anti-peptide antibodies can be used to detect peptides in the screening assays described herein. The antibody can optionally be linked 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.

[0441] Non-human antibodies against 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. Patent No. 6,602,503, EP 239 400, U.S. Patent No. 5,693,761, and U.S. Patent No. 6,407,213, the disclosures of each of which are incorporated by reference in their entirety.

[0442] Methods for Selecting a Therapy

[0443] A method for selecting a therapy for a subject having cancer (e.g., a plasma cell disorder such as multiple myeloma and / or Waldenstrom macroglobulinemia or any cancer in which XBP1, CD138, or CS1 is expressed (e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, leukemia such as AML or CML)) or a pre-cancerous condition (e.g., smoldering multiple myeloma) includes the steps of: optionally, determining that one or more cells (e.g., plasma cells) of the subject's cancer express XBP1; and if one or more cells express XBP1, selecting a peptide or composition described herein as a therapy for the subject, the peptide or composition e.g., an XBP1 peptide or composition comprising an XBP1 peptide described herein.

[0444] A method for selecting a therapy for a subject having cancer can include the steps of: optionally, determining that one or more cells (e.g., plasma cells) of the subject's cancer release and express CD138; and if one or more cells express CD138, selecting a peptide or composition described herein as a therapy for the subject, the peptide or composition e.g., a CD138 peptide or composition comprising a CD138 peptide described herein.

[0445] A method for selecting a therapy for a subject having cancer can include the steps of: optionally, determining that one or more cells (e.g., plasma cells) of the subject's cancer release and express CS-1; and if one or more cells express CS-1, selecting a peptide or composition described herein as a therapy for the subject, the peptide or composition e.g., a CS-1 peptide or composition comprising a CS-1 peptide described herein.

[0446] It is understood that when two or more of one or more cells (e.g., plasma cells) of a subject's cancer express XBP1, CD138, and CS-1, a suitable combination of peptides can be delivered to the subject, e.g., via the compositions described herein. For example, when it is determined that one or more cells (e.g., plasma cells) of a subject's cancer express XBP1 and CD138, a method of selecting a therapy can include selecting, for the subject, as a therapy: at least one XBP1 peptide and at least one CD138 peptide described herein or a composition comprising the peptides.

[0447] Methods for determining the release of one or more cells expressing XBP1, CD138, or CS-1 are known in the art and are described above. For example, a biological sample (e.g., a blood sample or a lymph node tissue sample) obtained from a subject can be assayed using an XBP1, CD138, or CS-1 specific antibody prepared by the methods described herein to detect the presence or amount of XBP1, CD138, or CS-1 polypeptide expressed by the cells (or cell lysates). (See, e.g., the Examples and Sambrook et al., supra). Methods for determining the presence or amount of a polypeptide in a biological sample include, for example, ELISA, immunohistochemistry, flow cytometry, Western blotting, or dot blot assays.

[0448] In some embodiments, any of the methods described herein can further include the step of providing and / or obtaining a biological sample from a subject. Suitable biological samples for the methods described herein include any biological fluid, cell, tissue, or portion thereof that includes an analyte protein of interest (e.g., an XBP1, CD138, or CS-1 protein). The biological sample can be, for example, a sample obtained from a subject (e.g., a mammal such as a human) or a sample that can be derived from the subject. For example, the sample can be a tissue section obtained by biopsy, or cells placed in or suitable for tissue culture. The biological sample can also be a cell-containing biological fluid such as urine, blood, plasma, serum, saliva, semen, sputum, cerebrospinal fluid, tears, mucus, or aspirate (e.g., lung or nipple aspirate), or such a sample adsorbed onto paper or a polymer matrix. The biological sample can be further fractionated (if needed) to a fraction containing a specific cell type. For example, a blood sample can be fractionated into serum or a fraction containing a specific blood cell type such as red blood cells or white blood cells (lymphocytes). If needed, the sample can be a combination of sample types from a subject such as a combination of tissue and biological fluid.

[0449] Biological samples can be obtained from a subject, e.g., a subject having cancer (e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia (e.g., AML or CML), multiple myeloma, and / or Waldenstrom macroglobulinemia), suspected of having cancer, or at risk of developing cancer. Any suitable method for obtaining a biological sample can be employed, although exemplary methods include, e.g., phlebotomy, swabbing (e.g., oral swabbing), aspiration, or fine needle biopsy procedures. Non-limiting examples of tissues amenable to fine needle aspiration include lymph nodes, lung, thyroid, breast, and liver. Samples can also be collected by, e.g., microdissection (e.g., laser capture microdissection (LCM) or laser microdissection (LDM)), bladder irrigation, smear (PAP smear), or catheter lavage.

[0450] After detecting cancer (e.g., lung cancer, liver cancer, cholangiocarcinoma, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, leukemia (e.g., AML or CML), multiple myeloma, and / or Waldenstrom macroglobulinemia) or a pre-cancerous condition such as smoldering multiple myeloma in a subject, e.g., using the methods described above, a physician (e.g., doctor) can select an appropriate treatment modality for the subject, e.g., by: (i) writing a medical prescription; (ii) giving (but not necessarily administering) a drug to the subject (e.g., giving a sample of a prescription drug to the patient when the patient is in the examining room); (iii) communicating with the patient (verbally, in writing (other than the prescription), or electronically (e.g., email, electronic posting to a secure site)) regarding the treatment modality recommended or suggested (e.g., a therapy comprising one or more of the peptides described herein); or (iv) identifying an appropriate treatment modality for the subject and disseminating the information to other medical channels, e.g., by way of a medical record. The latter (iv) may be useful, e.g., in situations where more than one therapy or therapeutic agent is to be administered to the patient by different physicians.

[0451] After detecting XBP1, CD138, or CS-1 in a subject (using any of the methods above); and / or selecting a therapy for the subject, a physician (e.g., doctor) can administer an appropriate treatment modality to the subject. Methods for administering the therapy include one or more of the peptides described herein detailed above.

[0452] In addition, a physician may alternatively prescribe and / or administer one or more additional therapeutic agents to treat cancer or one or more drugs to treat side effects of an anti-cancer agent. Suitable chemotherapeutic agents for treating multiple myeloma and / or Waldenstrom's macroglobulinemia include, for example, melphalan, cyclophosphamide, vinblastine, doxorubicin, prednisone, dexamethasone, proteasome inhibitors (e.g., bortezomib), thalidomide or lenalidomide.

[0453] Side effects of anti-cancer agents include, for example, anemia, gastrointestinal symptoms (e.g., nausea, vomiting, diarrhea), leukopenia (decrease in the number of white blood cells, which can lead to infection), temporary hair loss or thrombocytopenia (decrease in the number of platelets, which can lead to bleeding). Accordingly, a physician may prescribe or administer to a subject a chemotherapeutic agent such as vinblastine in combination with an anti-anemia drug such as epoetin alfa (e.g., or ).

[0454] Manufacturer's Kits and Articles

[0455] The present disclosure is also characterized by a variety of kits. The kits may include, for example, 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 kits may include one or more pharmaceutically acceptable carriers and / or one or more immunostimulants and / or one or more immunomodulators. The immunostimulant may be, for example, a T helper epitope, a modified peptide ligand or an adjuvant. In one embodiment, the immunostimulant may be a combination of carboxymethylcellulose, polyinosinic-cytidylic 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 a protein (e.g., a cytokine, complement, GCSF, GM-CSF). In one embodiment, the immunomodulator is a protein, e.g., an antibody that 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).

[0456] The kit may further comprise one or more therapeutic, diagnostic, or prophylactic agents. The one or more therapeutic, diagnostic, or prophylactic agents include, but are not limited to: (i) agents that modulate the inflammatory response (e.g., aspirin, indomethacin, ibuprofen, naproxen, steroids, cromolyn sodium, or theophylline); (ii) agents that affect renal and / or cardiovascular function (e.g., furosemide, thiazides, amiloride, spironolactone, captopril, enalapril, lisinopril, nifedipine, verapamil, digoxin, isosorbide dinitrate, dobutamine, lidocaine, quinidine, adenosine, digitalis, mevastatin, simvastatin, lovastatin, or mevalonic acid); (iii) drugs that affect gastrointestinal function (e.g., omeprazole or sucralfate); (iv) antibiotics (e.g., tetracycline, clindamycin, amphotericin B, quinine, methicillin, vancomycin, penicillin, 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, bleomycin, doxorubicin, mitomycin C, hydroxyurea, prednisone, tamoxifen, cisplatin, or dacarbazine); (vi) immunomodulators (e.g., interleukins, interferons (e.g., interferon γ (IFN-γ), granulocyte macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor α (TNFα), tumor necrosis factor β (TNFβ), cyclosporine, FK506, azathioprine, steroids); (ix) drugs that act on blood and / or 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, estrogen, androgen, progesterone, gonadotropin-releasing hormone (GnRH), thyroxine, triiodothyronine); hormone antagonists; agents that affect calcification and bone turnover (e.g., calcium, phosphate, parathyroid hormone (PTH), vitamin D, bisphosphonates, calcitonin, fluoride).

[0457] Another feature is a manufacturer's product comprising: 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 inducing an immune response in a mammal (e.g., any of the mammals described herein). The label may further indicate that the composition is to be administered to a mammal having cancer, suspected of having cancer, or at risk of developing cancer, such as lung cancer, liver cancer, bile duct cancer, gastric cancer, cervical cancer, nasopharyngeal cancer, breast cancer, colon cancer, pancreatic cancer, prostate cancer, multiple myeloma, smoldering multiple myeloma, and / or Waldenstrom macroglobulinemia. The composition of the manufacturer's product may be dried or lyophilized, and it may include, for example, one or more solutions (and / or instructions) for dissolving the dried or lyophilized composition.

[0458] The manufacturer's product may further include instructions (e.g., as described above) for administering the composition to a mammal.

[0459] The following examples are intended to illustrate and not limit the invention. Examples

[0460] Example 1: Materials and Methods

[0461] Cell LinesMultiple 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 a gift from Dr. Y. Matsuo, Fujisaki Cell Center, Okayama, Japan. The T2 cell line, which is a human B cell and T cell hybrid expressing the HLA-A2.1 molecule (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, TX) and used as a source of antigen-presenting cells (APCs). K562-A*0201 cells were provided by Karen Anderson (Dana Farber Cancer Institute, Boston, MA) and used in immunomonitoring assays to present individual peptides to CTLs. Multiple 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 μg / ml streptomycin (Gibco-Life Technologies).

[0462] Reagents Mouse anti-human CD80 or CD83 monoclonal antibodies (mAbs) conjugated to phycoerythrin (PE) were purchased from Immunotech (Hialeigha, FL). Mouse anti-human CD3, CD4, CD8, CCR7, CD45RO, CD69, CD107α, IFN-γ, and HLA-A2 mAbs conjugated to 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, CA). Recombinant human IL-2, IL-4, IFN-, and TNF- were purchased from R&D Systems (Minneapolis, MN) and GM-CSF was obtained from Immunex (Seattle, WA).

[0463] Synthetic Peptides Influenza virus matrix peptide 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 natural unspliced XBP1 peptides were designed: XBP1 118-126 (LLREKTHGL; SEQ ID NO:1); XBP1 185-193 (NISPWILAV(SEQ ID NO:2)); XBP1 190-198 (ILAVLTLQI(SEQ ID NO:3)); XBP1 193-201 (VLTLQIQSL(SEQ ID NO:4)); XBP1 111-119 (KLLLENQLL(SEQ ID NO:5)); XBP1 94-102 (RMSELEQQV(SEQ ID NO:27)); including SP XBP1 197-205 (GILDNLDPV(SEQ ID NO:7)); SP XBP1 194-202 (ILLGILDNL(SEQ ID NO:8)); SP XBP1 368-376 (ELFPQLISV(SEQID NO:9)) of three natural spliced XBP1 peptides; aberrant XBP1 (YISPWILAV(SEQ ID NO:6)); and aberrantly spliced XBP1 (YILDNLDPV(SEQ ID NO:24)); and YLFPQLISV(SEQ ID NO:10)) peptides were examined as possible HLA-A2 binding peptides. As used herein, "aberrant" (e.g., aberrant peptide) refers to a form of such a peptide in which one or more amino acids have been modified from the wild-type or original sequence to produce a peptide that is more immunogenic than the corresponding wild-type peptide. For example, in the exemplary aberrant peptides described above, the bolded amino acids indicate amino acids that have been diluted from the wild-type sequence of XBP1.

[0464] Four natural CD138 peptides were designed: CD138 256-264 (VIAGGLVGL(SEQ ID NO:11)); CD138 260-268 (GLVGLIFAV(SEQ ID NO:12)); CD138 5-13 (ALWLWLCAL(SEQ ID NO:13)); and CD138 7-15(WLWLCALAL (SEQ ID NO:14)) and examined as a potential HLA-A2 binding peptide.

[0465] Four natural CS1 peptides were designed: CS1-P1: CS1 236-245 (LLLSLFVLGL (SEQ ID NO:15)); CS1-P2: CS1 239-247 (SLFVLGLFL (SEQ ID NO:16)); CS1-P3: CS1 232-240 (LLVPLLLSL (SEQ ID NO:17)); and CS1-P4: CS1 9-17 (TLIYILWQL (SEQ ID NO:18)) (using three different databases, RANKPEP, BIMAS, and NetMHC) and examined as a potential HLA-A2 binding peptide. (See, e.g., Reche et al. (2002) Human Immunology 63:710 - 709).

[0466] XBP-1 and CD138 peptides were synthesized by standard FMOC (9-fluorenylmethyl-oxycarbonyl) chemistry (Biosynthesis, Lewisville, TX), purified to >85% by reverse-phase chromatography, and the molecular weights were verified by mass spectrometry. CS1 peptides were synthesized by New England Peptides LLC with a purity higher than 95%.

[0467] Aberrant XBP1 US 185-193 (YISPWILAV) (SEQ ID NO:6), aberrant XBP1 SP 368-376 (YLFPQLISV) (SEQ ID NO:10), native CD138 260-268 (GLVGLIFAV) (SEQ ID NO:12) and native CS1 239-247 (SLFVLGLFL) (SEQ ID NO:16) peptides are derived from XBP1 unspliced (US), XBP1 spliced (SP), CD138, and CS1 antigens, respectively. Influenza virus matrix protein was selected 58-66(GILGFVFTL)(SEQ ID NO:25) and CMV pp65 (NLVPMVATV)(SEQ ID NO:28) were used as HLA-A2 specific control peptides. All peptides were synthesized by standard fmoc (9-fluorenylmethyl-oxycarbonyl) chemistry, purified to >90% by reverse-phase chromatography, and their molecular weights were verified by mass spectrometry (Biosynthesis, Lewisville, TX). The lyophilized peptides were dissolved in DMSO (Sigma, St. Louis, MO), diluted in AIM-V medium (Gibco-Life Technologies) and stored at -140 °C.

[0468] Peptide Binding Assay The binding affinity of a mixture of four HLA-A2 peptides (irregular XBP1US 185-193 , irregular XBP1 SP 368-376 , CD138 260-268 and CS1 239-247 ) was evaluated 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 1x10 6 cells / ml, and transferred to 48-well tissue culture plates. Cells were pulsed with a mixture of four peptides with a total peptide concentration range of 0 - 50 μg / ml plus 3 μg / ml human β2-microglobulin (Sigma, St Louis, MO) and incubated at 37 °C, 5% CO 2 in humidified air. After overnight incubation, the cells were washed, stained with mouse anti-human HLA-A2-FITC mAb for 15 minutes at 4 °C, and analyzed using a FACSCanto TM II flow cytometer (Becton Dickinson, San Jose, CA).

[0469] Peptide Stability Assay The HLA-A2 stability of the multiple peptide mixture was examined over time. After overnight incubation of T2 cells pulsed with the multiple peptide mixture (25 μg / ml; 6.25 μg / ml / peptide), the cells were washed to remove unbound peptides and incubated with 10 μg / ml brefeldin A (Sigma) at 37 °C and 5% CO 2 for 1 hour to block the cell surface expression of newly synthesized HLA-A2 molecules. At 0, 2, 4, 6, and 14 hours after BFA treatment, the peptide / HLA-A2 complex stability was measured by staining the cells with mouse anti-human HLA-A2-FITC mAb and analyzing by flow cytometry.

[0470] Generation of Monocyte-Derived Mature Dendritic Cells In Ficoll-Paque TM Plus (Amersham Pharmacia Biotech AB, Uppsala Sweden), peripheral blood mononuclear cells (PBMCs) were isolated from leukopaks obtained from HLA-A2 + normal individuals by standard density gradient centrifugation. To generate dendritic cells (DCs), monocytes isolated as the adherent cell fraction were cultured for 7 days in RPMI-1640 medium (Gibco-Life Technologies) supplemented with 10% FCS in the presence of 1,000 U / ml GM-CSF and 1,000 U / ml IL-4. Fresh medium supplemented with GM-CSF and IL-4 was added to the cultures every other day. On day 7, mature DCs (mDCs) were obtained by adding 1,000 U / ml IFN-α and 10 ng / ml TNF-α in 10% FCS-RPMI, along with fresh GM-CSF and IL-4, and incubating for an additional 3 days.

[0471] CD3 + Isolation of T cells By using magnet and CD3 + T cells were obtained from the non-adherent cell fraction by negative selection. Briefly, T cell enrichment was accomplished by depleting non-CD3 T cells (including B cells, monocytes, NK cells, erythroid cells, platelets, and basophils) via labeling with bispecific tetramer antibody complexes directed against CD14, CD16, CD19, CD20, CD36, CD56, CD66b, CD123, and glycophorin A.

[0472] Isolate primary CD138 from the bone marrow mononuclear cells of MM patients + cells In Ficoll-Paque TM Plus, bone marrow monocytes (BMMCs) were isolated from bone marrow cells obtained from MM patients by standard density gradient centrifugation. CD138 MM cells were isolated from BMMCs using + CD138-positive immunomagnetic selection technology (StemCell Technologies).

[0473] Induction of Peptide-Specific CTLs By +Repeated stimulation of CD3+ T lymphocytes from HLA-A24+ donors generated CTLs specific for a single peptide (peptide-specific CTLs) or CTLs specific for multiple peptides (MP-CTLs) in vitro (see Figure 25 ). Briefly, the cells were cultured at 37°C and 5% CO in a humidified atmosphere. 2 Using either individual peptides or irregular XBP1US 185-193 , irregular XBP1 SP 368-376 、CD138 260-268 and CS1 239-247 APCs (mDCs or T2 cells) were pulsed with a mixture of peptides (25 μg / ml total peptide). Loaded APCs were collected, washed, irradiated at 20 Gy and resuspended in AIM-V medium supplemented with 10% human AB serum. Using irradiated mp-pulsed mDCs, CD3 + T cell ratio priming autologous CD3 + T cells. Cultures were restimulated with irradiated APC / mp every 7 days for a total of 4 cycles to generate CTLs specific for mp. IL-2 (50 U / ml) was added to the cultures 2 days after the second stimulation and supplemented until the culture was completed.

[0474] Phenotypic Analysis of XBP1-CTLs, CD138-CTLs, or Target Cells One week after the fourth stimulation, total CD3 T cells of MP-CTLs and control cells were evaluated by staining with CD3-PacBlue, CD8-APC-H7, CCR7-PeCy7, CD45RO-PE, and / or CD69-PerCP mAbs for 30 min at 4°C. + CD8 + T cells or untreated, effector memory, and activated CD3 + CD8 + T cells. After staining, cells were washed, fixed with 2% paraformaldehyde-PBS and analyzed by flow cytometry.

[0475] Western BlotApproximately 100 μg of protein lysates from each cell line (U266, McCAR, ML-2, and MM1S) were suspended in Laemmli sample buffer (0.1 M Tris-HCl buffer, pH 6.8, containing 1% sodium dodecyl sulfate (SDS), 0.05% β-mercaptoethanol, 10% glycerol, and 0.001% bromophenol blue), boiled for 2 minutes, and subjected to 8 - 16% gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at 80 V for 2 hours (Xcell Surelock Mini Cell, Invitrogen, Carlsbad, CA). A protein ladder (a mixture of proteins of known molecular weights) was used as a size marker in the gel to determine the molecular weight of the peptides (Invitrogen, Carlsbad, CA). The gel was electroblotted onto a nitrocellulose membrane (Trans-Blot, 0.2 μm transfer membrane, Bio-Rad Laboratories, CA) at 40 V in Tris-glycine buffer for 2 hours. The transfer of proteins onto the nitrocellulose membrane was confirmed by Ponceau S staining. The membrane was incubated with mouse anti-human XBP1 antibody or anti-human CD138 antibody in phosphate-buffered saline and Tween 20 containing 1% BSA (PBST) for 1 hour with continuous shaking. The membrane was washed three times with PBST and incubated in anti-mouse IgG-horseradish peroxidase conjugate in PBST containing 3% non-fat milk powder for 1 hour. After washing, specific proteins were detected using enhanced chemiluminescence according to the instructions provided in the product manual (Amersham Life Sciences Inc., Arlington Heights, IL).

[0476] IFN-γ ELISA The IFN-γ release of XBP1-CTL, CD138-CTL, or CS1-CTL after co-culture with multiple myeloma (MM) cells (McCAR, MM1S), acute myeloid leukemia (AML) cells (ML-2), or T2 cells (described above) was measured using a human IFN-γ ELISA kit from BD Biosciences (San Diego, CA) (see Figure 27)。Briefly, a dilution of purified IFN-γ as a standard or CTL supernatant was transferred into wells of a 96-well plate pre-coated with a monoclonal anti-human IFN-γ capture antibody and incubated for 2 hours at room temperature. After several washes, a buffer containing the detection antibody and avidin-horseradish peroxidase conjugate was added to each well and incubated for 1 hour at room temperature. The wells were washed and a horseradish peroxidase substrate solution was added to each well and incubated for 30 minutes at room temperature. A stop solution was added to each well and the absorbance at 450 nm was determined using a PerkinElmer Wallac Victor2 counter (PerkinElmer, Wellesley, MA). The amount of cytokine present in the CTL culture supernatant was calculated based on the IFN-γ standard curve.

[0477] Cell Proliferation Traced by Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) CTL proliferation was measured after co-culture with multiple myeloma (MM) cells (McCAR, MM1S), acute myeloid leukemia (AML) cells (ML-2), or T2 cells (described above) (see Figure 27 and 32 ). Individual XBP1-CTL, CD138-CTL, CS1-CTL, or multi-peptide-generated CTLs were washed twice in PBS (Gibco-BRL) and resuspended in RPMI-1640 medium at a concentration of 1 x 10 6 cells / ml. CFSE (Molecular Probes, Eugene, OR) in the form of a 5 mM stock solution in DMSO was added to the CTLs to a final concentration of 5 μM and incubated for 10 minutes at 37 °C in a CO 2 incubator in the dark. After incubation, a volume of ice-cold PBS (containing 2% FCS) equal to 5 times the volume of the CTL cells was added to the cells to quench the reaction. The cells were incubated on ice for 5 minutes, centrifuged, and after washing three times, resuspended in fresh PBS (containing 2% FCS). The CFSE-labeled T cells were adjusted to a concentration of 2 x 10 6 cells / ml using RPMI medium and stimulated with 2 x 10 5 cells / ml primary multiple myeloma cells, MM cell lines, various cancer cell lines, or K562-A2 cells (which present individual peptides). The stimulated CFSE-labeled cells were examined by flow cytometry.

[0478] Cytotoxicity AssayAs described by Roden et al. (1999) J. Immunol Methods 226:29-41, the cytotoxic activity of XBP1-CTL, CD138-CTL, CS1-CTL, or CTL generated by multiple peptides alone was measured by calcein release assay. Briefly, target cells (3x10 5 cells) (including T2, U266 cells, McCAR cells, ML-2 cells, MM1S cell line, or primary multiple myeloma cells) were incubated in serum-free medium containing 10 mM calcein-AM (Molecular Probes) at 37 °C for 30 minutes, washed three times in PBS with 5% FCS, and incubated with effector cells (5x10 3 cells / well) at various effector:target cell ratios in 96-well U-bottom microtiter plates (three replicate wells / sample). The plates were incubated at 37 °C and 5% CO 2 for 3 hours. After incubation, the cells were pelleted by centrifugation at 1,000 rpm for 5 minutes and 100 μl of supernatant was transferred to wells of a 96-well flat-bottom microtiter plate (Nunc), and calcein release was measured as the amount of fluorescence released from the cells (using VICTOR 2 -1420 multilabel counter (PerkinElmer, Boston, MA)). Maximum calcein release was obtained from detergent-released target cell counts, and spontaneous release was obtained from target cell counts in the absence of effector cells. Cytotoxicity was calculated as follows: % specific lysis = [(experimental release – spontaneous release) ÷ (maximum release – spontaneous release)] x 100.

[0479] CD107α Degranulation Assay CD107α degranulation was measured after co-culture with multiple myeloma (MM) cells (McCAR, MM1S), acute myeloid leukemia (AML) cells (ML-2), or T2 cells (above) (see Figure 27 and 30)。As described by Betts et al. (2003) and Mittendorf et al. (2005), with minor modifications (detailed below), a CD107α degranulation assay was performed, which is a measure of cytotoxic activity. Primary multiple myeloma cells, MM cell lines, various cancer cell lines, or K562-A2 cells pulsed with individual peptides were co-cultured with CTLs at various effector:target ratios. 10 μl aliquots of each of CD107α and CD107b (both conjugated to the detectable marker FITC) were added to each well, along with CD138-CTL. The cell-containing plates were centrifuged at 1000 rpm for 5 minutes and incubated at 37 °C for 1 hour. After incubation, brefeldin A and monensin were added to each well and the cells were incubated for a further four hours at 37 °C. The cells were collected and washed and stained with fluorescent dye-conjugated anti-human MABs. The cells were washed and analyzed by flow cytometry.

[0480] CD107α Upregulation and Intracellular IFN-γ Production After co-culture with multiple myeloma (MM) cells (McCAR, MM1S), acute myeloid leukemia (AML) cells (ML-2), or T2 cells (above), CD107α upregulation and IFN-γ production were measured (see Figure 27 and 30 ). CD107α degranulation and IFN-γ-producing CD8 + CTLs were determined by flow cytometry by staining for cell surface markers and intracellular cytokines. Briefly, cells were stimulated (HLA-A2 6 McCAR or U266 MM cell lines or K562-A*0201 cells pulsed with the appropriate peptide) with 1x10 + stimulator cells and 1x10 6 responder cells (MP-CTL, control T cells). CD107α mAb was added to the cultures and the cells were placed at 37 °C, 5% CO 2In an incubator. After 1 hour of incubation, CD28 / CD49d mAb (BD), the protein transport inhibitor brefeldin A (BD), and monensin (BD) were added to the cell culture and incubated for another 5 hours. As a baseline control, MP-CTLs were cultured in a medium with CD28 / CD49d mAb, brefeldin A, and monensin without additional stimulation. After incubation, the cells were harvested, washed, and stained with CD3-PacBlue and CD8-APC-H7, CCR7-PeCy7, CD45RO-PE, and / or CD69-PerCP anti-human mAbs for 30 minutes. Then the cells were permeabilized, fixed using Cytofix / Cytoperm (BD), and stained with anti-IFN-γ FITC mAb for 45 minutes to detect intracellular cytokine production. Finally, the cells were washed with Perm / Wash solution (BD) and fixed in 2% paraformaldehyde and captured by flow cytometry.

[0481] IL-2 Production Assay CD107α degranulation and IL-2-producing CD3+CD8+ CTLs were identified by flow cytometry through cell surface marker and intracellular cytokine staining. Briefly, peptide-specific CTLs or control T cells were stimulated with each specific stimulator in the presence of CD107a mAb. After 1 hour of incubation, CD28 / CD49d mAb (BD) and the protein transport inhibitors brefeldin A and monensin were added to the culture and incubated for another 5 hours. As a baseline control, CTLs were cultured in a medium containing only CD28 / CD49d mAb, brefeldin A, and monensin. After incubation, the cells were stained with CD3-PacBlue and CD8-APC-H7 anti-human mAbs, then fixed / permeabilized and stained with anti-IL-2 APC anti-human mAb to detect intracellular cytokine production. After staining, the cells were washed three times with Perm / Wash solution, fixed in 2% paraformaldehyde and analyzed by flow cytometry.

[0482] Statistical Analysis Results are presented as mean ± SE. Unpaired Student's t-test was used to compare groups. Differences were considered significant when p < 0.05.

[0483] Example 2: Multipeptide (MP) Mixtures of Unspliced XBP1, Spliced XBP1, CD138, and CS1-Specific Peptides Exhibiting high HLA-A2 binding affinity and stability

[0484] Four immunogenic peptides were demonstrated individually: the truncated XBP1 US 185-193 (YISPWILAV, SEQ ID NO:6), the truncated XBP1 SP 368-376 (YLFPQLISV, SEQ ID NO:10), native CD138 260-268(GLVGLIFAV, SEQ ID NO:12) and native CS1 239-247 (SLFVLGLFL, SEQ ID NO:16) (Table 1) induces an immune response. Here we evaluated it as an MP mixture. The HLA-A2-specific binding and stability of the MP mixture were evaluated by measuring the upregulation of HLA-A2 molecules on T2 cells using flow cytometry (27). Peptide binding assays showed that the mean fluorescence intensity (MFI) of HLA-A2 on T2 cells increased in a dose-dependent manner (0 - 50 μg / ml) and reached a plateau at a total peptide concentration of 25 μg / ml (6.25 μg / peptide / ml; MFI: 10,787.33 ± 2,371.71), similar to that at the highest total peptide concentration of 50 μg / ml (MFI: 10,889.33 ± 2,888.48)( Figure 1a ). Therefore, an MP concentration of 25 μg / ml (6.25 μg / peptide / ml) was chosen to evaluate HLA-A2 binding stability.

[0485] Figure 1b In the peptide binding stability assay, T2 cells were pulsed overnight with a 25 μg / ml MP mixture, washed to remove unbound peptides, and then treated with brefeldin A (BFA) to block cell surface expression of newly synthesized HLA-A2 molecules. The HLA-A2 MFI of T2 cells was then evaluated at 0, 2, 4, 6, or 14 hours after BFA treatment. Flow cytometry analysis showed that the stability of the MP mixture was highly maintained up to 6 hours after BFA treatment (MFI: 0 hours = 9,726.00 ± 1,373.24, 2 hours = 9,132.33 ± 1,435.51, 4 hours: 9,125.33 ± 1,130.62, 6 hours: 8,818.67 ± 413.50)( 58–66 At 14 hours after BFA treatment, the HLA-A2-specific affinity of the MP mixture was lower but greater than (MFI: 6,793.67 ± 1,617.01) that of the control influenza virus matrix protein (IVMP)

[0486] Table 4: Evaluation of natural and degenerate epitopes with multi-peptides targeting MM

[0487]

[0488] Example 3: Multi-peptide specific CTLs exhibit different phenotypes presenting specific T cell subsets

[0489] peptide affinity (MFI: 4,921.33 ± 1,428.16). Based on these results, we confirmed the high level of HLA-A2-specific affinity and stability of the MP mixture and proceeded to further evaluate the immunogenicity of the mixture and its ability to induce MM-specific CTLs.Stimulation of HLA-A2 by APC pulsed weekly with MP mixture (total 25 g / ml; 6.25 g / ml / peptide) + T cells from normal donors were used to generate MP-CTLs. One week after the fourth stimulation, the phenotype and functional activity of the resulting MP-CTLs were evaluated. Flow cytometry analysis showed that compared to control T cell cultures (donor 1: 25%, donor 2: 25%; Figure 2 ), MP-CTLs contained a higher proportion of CD3 + CD8 + T cells (donor 1: 86%, donor 2: 74%). We also observed distinct phenotypic changes in the CD3 + CD8 + T cell subset in MP-CTLs. The frequency of effector memory T cells (EM: CD45RO + CCR7 - / CD3 + CD8 + ) was increased (donor 1: control 5% vs. MP-CTL 44%, donor 2: control 4% vs. MP-CTL 35%), which was associated with a corresponding decrease in untreated T cells (CD45RO - CCR7 + / CD3 + CD8 + ) (donor 1: control 74% vs. MP-CTL 8%, donor 2: control 60% vs. MP-CTL 6%). In addition, we observed an increased frequency of activated CD69 + / CD3 + CD8 + T cells in MP-CTLs compared to control T cell cultures (donor 1: control 3% vs. MP-CTL 39%, donor 2: control 5% vs. MP-CTL 13%; Figure 2 ). Thus, these results indicate that repeated stimulation of CD3 + T cells with MP mixtures specific for XBP1, CD138, or CS1 leads to distinct phenotypic alterations and expansion of the antigen-specific CTL characteristic CD3 + / CD8 + T cell subset.

[0490] Example 4: Multipeptide-specific CTLs include a high proportion of CD8 cells that respond to HLA-A2 and produce IFN-γ against MM cells + CD8 cells producing IFN-γ against MM cells + CTL

[0491] The ability of MP-CTLs to produce intracellular IFN-γ after stimulation with HLA-A2 + MM cell lines was analyzed by flow cytometry. In response to HLA-A2 + MM cell lines, EM (CD45RO+ CCR7 - ) and activated (CD69 + )CD3 + CD8 + T cells produce IFN-γ ( Figure 3 The frequency of IFN-γ producing cells was increased after stimulation with 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]. When stimulated with MM cell lines, the frequency of untreated (CD45RO - CCR7 + )CD3 + CD8 + T cells showed minimal levels of IFN-γ production.

[0492] Example 5: Multiple peptide-specific CTLs show response to HLA-A2 + Cell proliferation of MM cells

[0493] The function of MP-CTL was analyzed using CFSE proliferation assay. + After stimulation of MM primary cells or cell lines, MP-CTL proliferation was measured as the decrease in fluorescence of CFSE-labeled MP-CTL (Q1-gated cells) on day 5 ( Figure 4 ). MP-CTL showed responses obtained from three different HLA-A2 + CD138 in MM patients + High levels of cell proliferation of primary cells (proliferating cells: 33%, 29% or 41%). In addition, MP-CTLs showed high levels of response to HLA-A2 + Cell proliferation of MM cell lines, the HLA-A2 + MM cell lines included McCAR (proliferating cells: 57%) and U266 (proliferating cells: 49%). MP-CTL cultured in medium alone showed a low level of proliferation (5%). In summary, these data suggest that when HLA-A2 + The functional activity of MP-CTLs was demonstrated by their proliferative capacity upon stimulation of primary MM cells or MM cell lines.

[0494] Example 6: Specific lysis of HLA-A2 + MM cells by multi-peptide specific CTL

[0495] We evaluated the cytotoxic activity of MP-CTL using a 4-hour calcein release assay. The cytotoxic activity of MP-CTL generated from CD3 + T cells from different HLA-A2 + donors against HLA-A2 + MM primary cells or cell lines ( Figure 5 ) was evaluated. HLA-A2 + primary MM cells were efficiently lysed by 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 exhibited high levels of cytotoxic activity against 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 different effector:target cell ratios. Compared to MP-CTL, control CD3 + T cells from the same donors showed significantly lower levels of cytotoxicity against HLA-A2 + MM primary cells or cell lines. In addition, MP-CTL did not lyse antigen-mismatched or MHC-mismatched tumor cells, including HLA-A2 + breast cancer cell line (MCF-7), HLA-A2 - MM cell line (MM1S) or HLA-A2 - primary cells from three different MM patients (data not shown). Collectively, these data confirm the HLA-A2-restricted and antigen-specific cytotoxic activity of MP-CTL.

[0496] Example 7: Multi-peptide specific CTLs generate individual immune responses against each relevant peptide

[0497] The ability of MP-CTL to degranulate (CD107α expression) and produce intracellular IFN-γ in response to each relevant peptide was analyzed, and the relevant peptides included the aberrant XBP1 US 185-193 (YISPWILAV) (SEQ ID NO:6), the aberrant XBP1 SP 368-376 (YLFPQLISV) (SEQ ID NO:10), the native CD138 260-268 (GLVGLIFAV) (SEQ ID NO:12) and the native CS1 239-247(SLFVLGLFL) (SEQ ID NO:16). Analysis was performed by measuring the MP-CTL responses specific for K562-A*0201 cells stimulated with the respective peptides. As controls, we used K562-A*0201 cells without peptide pulsing or K562-A*0201 cells pulsed with an irrelevant HLA-A2-specific CMV pp65 (NLVPMVATV) peptide (SEQ ID NO:28). Figure 6a Representative flow cytometric analysis of peptide-specific responses of MP-CTLs from donor A is shown. MP-CTLs showed a high proportion of responses to the XBP1 US (2.7%), CD138 (1.7%), and CS1 (12.5%) peptides, but not to the CD107α of the XBP1 SP (0.2%) peptide. + IFNγ + / CD3 + CD8 + T cells (gated Q2). No responses were observed for the irrelevant CMV pp65 peptide (0.2%) or the no-peptide control (0.2%). Additional analysis of MP-CTLs generated from three additional HLA-A2 + donors (donor B, donor C, donor D) was performed regarding their CD107α degranulation or IFN-γ production in response to K562-A*0201 cells Figure 6b presenting each individual peptide. Specific responses to all relevant peptides, but not to the irrelevant CMV pp65 peptide, were detected in MP-CTLs generated from each of these donors. However, variations were detected in the levels of specific responses in degranulation and IFN-γ production for each relevant peptide among CTLs generated from different individuals. Thus, these studies demonstrate that MP mixtures including XBP1US, XBP1 SP, CD138, and CS1 epitopes are able to induce responses against the respective peptides using specific CTLs that can target multiple antigens on MM cells.

[0498] Example 8: XBP1 expression in cancer cell lines

[0499] Expression of unspliced and spliced XBP1 antigens in different cancer cell lines was analyzed by flow cytometry. Relative expression levels are indicated by plus or minus signs and also numbers indicating the number of plus signs ( Figure 7 ).

[0500] Example 9: Proliferation of XBP1-CTLs in response to HLA-A2+ breast cancer cells

[0501] Proliferation of XBP1-CTLs in response to stimulator cells from breast cancer cell lines was analyzed using a CFSE proliferation assay. In HLA-A2 +Breast cancer cell lines MCF-7, HLA-A2 - Breast cancer cell line BT434, HLA-A2 + Prostate cancer cell line LnCap, HLA-A2 - Proliferation of XBP1-CTL was measured by the decrease in fluorescence of CFSE-labeled MP-CTL (cells gated in P3) on days 6 and 7 after prostate cancer cell lines VCap and NK-sensitive CML K562 or without cell stimulation. XBP1-CTL showed high levels of cell proliferation on day 6 ( Figure 8a ) and day 7 ( Figure 8b ) in response to HLA-A2 + breast cancer cells rather than other cancer cell lines that served as controls.

[0502] Example 10: IFN-γ production and cell activation of XBP1-CTLs in response to HLA-A2+ breast cancer cells

[0503] IFN-γ expression and CD69 upregulation of XBP1-CTL in response to breast cancer cell lines (MB231, MCF-7, BT434) and prostate cancer cell lines (LnCAP, VCap) were analyzed by flow cytometry. IFN-γ expression and activation (CD69) of XBP1-CTL were increased after stimulation with HLA-A2+ breast cancer cells (including MB231 and MCF-7) rather than other cancer cells that served as controls ( Figure 9 ).

[0504] Example 11: Degranulation (CD107α) of XBP1-CTLs in response to HLA-A2+ breast cancer cell lines

[0505] The ability of XBP1-CTL to degranulate in response to breast cancer cell lines (MB231, MCF-7, BT434) and prostate cancer cell lines (LnCAP, VCap) was analyzed. CD107α upregulation was analyzed by flow cytometry in gated CD8 + T cells as a measure of cytotoxic activity. A significant level of degranulation (CD107α upregulation) was detected in response to HLA-A2+ breast cancer cells (including MB231 and MCF-7) rather than other cancer cell lines that served as controls ( Figure 10 ).

[0506] Example 12: Proliferation of XBP1-CTLs in response to HLA-A2+ pancreatic cancer cell lines and colon cancer cell lines

[0507] Proliferation of XBP1-CTL in response to pancreatic cancer cell lines and colon cancer cell lines was performed using a CFSE-based assay. After using HLA-A2 + prostate cancer cell line (LnCap), HLA-A2 + pancreatic cancer cell line (8902), HLA-A2 - pancreatic cancer cell line (MiaPaca), HLA-A2+ Colorectal cancer cell line (LS180) and HLA-A2 - After stimulation with the colorectal cancer cell line (WiDr) or cell-free control, the proliferation of XBP1-CTL was measured by the decrease in fluorescence of CFSE-labeled XBP1-CTL on day 6. XBP1-CTL showed a response to HLA-A2 + Pancreatic cancer cell line 8902 and HLA-A2 + Higher levels of cell proliferation in the colorectal cancer cell line LS180( Figure 11 )

[0508] Example 13: IFN-γ production of XBP1-CTLs in response to HLA-A2+ pancreatic cancer cell lines and colon cancer cell lines and degranulation

[0509] The ability of XBP1-CTL to degranulate and produce IFN-γ in response to pancreatic or colorectal cancer cell lines was analyzed. XBP1-CTL was incubated with the following: HLA-A2 + pancreatic cancer cell lines (Pan1 and PL45), HLA-A2 - pancreatic cancer cell line (MiaPaca), HLA-A2 + colorectal cancer cell lines (LS180 and SW480), HLA-A2 - prostate cancer cell line (WiDr) or cell-free control. CD107α and IFN-γ production were analyzed by flow cytometry. CD107α upregulation and IFN-γ production of XBP1-CTL were increased in stimulation with HLA-A2 + pancreatic cancer cells and HLA-A2 + colorectal cancer cells rather than with other control cancer cells, and showed responses in an antigen-specific and HLA-A2-restricted manner( Figure 12 )

[0510] In addition, as shown by retrieving the public databases of canEvolve( http: / / www.canevolve.org / AnalysisResults / AnalysisResults.html ) and Oncomine( http: / / www.webcitation.org / getfile?fileid=b cbe297e4085b19933cca759a88e0e2b9fac3b1e ), it was found that the XBP1 gene expression in primary tumor cells from breast cancer or colorectal cancer patients was significantly higher than that in cells from healthy donors, as shown below:

[0511] canEvolve

[0512]

[0513]

[0514] Oncomine

[0515]

[0516] Example 14: CD138 and CS1 expression in cancer cell lines

[0517] The CD138 and CS1 antigen expressions of different cancer cell lines were analyzed by flow cytometry. For different cancer cell lines, the relative CD138 and CS-1 expression levels were indicated by a minus or plus sign and also by numbers indicating the expression levels of CD138 ( Figure 13 ) and CS1 ( Figure 14 ).

[0518] Example 15: Percentage of CD8+ CTLs in T cells from relapsed multiple myeloma patients stimulated with XBP1 / CD138 / CS1 peptides Example 16: Proliferation of MP-CTLs from relapsed multiple myeloma patients

[0519] The specific T cells of multiplex peptide-specific CTLs (MP-CTLs) generated from T cells obtained from four HLA-A2+ smoldering multiple myeloma patients were analyzed by flow cytometry. The MP-CTLs generated from the T cells of each patient showed increased percentages of CD3+CD8+ CTLs and decreased percentages of CD3+CD4+ Th cells (Figure 15).

[0520] Figure 16a

[0521] The proliferation of MP-CTLs generated from two smoldering multiple myeloma patients in response to different MM cell lines was analyzed using a CFSE-based assay. In the presence or absence of stimulation with HLA-A2 + cell line (McCAR), HLA-A2 - cell line (MM1S, RPMI), NK-sensitive cell line (K562), cell proliferation was measured by the decrease in fluorescence of CFSE-labeled MP-CTLs (P3 gated) on days 5, 6, and 7. The MP-CTLs generated from the T cells of two SMM patients showed high levels of cell proliferation in response to the HLA-A2 + multiple myeloma cell line McCAR but not in response to HLA-A2- cell lines or NK-sensitive cell lines, indicating HLA-A2-restricted MP-CTL activity against multiple myeloma cells ( Example 17: IFN-γ production of MP-CTLs generated from relapsed multiple myeloma patients , b).

[0522] Figure 17a

[0523] The IFN-γ production of MP-CTLs generated from four smoldering multiple myeloma patients in response to different MM cell lines was analyzed. After stimulation with HLA-A2+ cell lines (McCAR, U266), HLA-A2- cell lines (MM1S, RPMI), NK-sensitive cell line (K562) or without any stimulation, production was analyzed by flow cytometry. The MP-CTLs generated from the four patients showed increased IFN-γ production in an HLA-A2-restricted manner after stimulation with HLA-A2 + multiple myeloma cell lines (Example 18: Degranulation (CD107α) of MP-CTLs generated from relapsed multiple myeloma patients against myeloma cell lines , b).

[0524] Figure 18b Example 19: Percentage of IFN-γ+CD107α+ double positive cells in the CD3+CD8+CD137+ subset of MP-CTLs generated from relapsed multiple myeloma patients

[0525] The degranulation of HLA-A2+ and HLA-A2- cancer cell lines with different MP-CTL responses generated from four smoldering multiple myeloma patients was analyzed as a measure of cytotoxic activity. The MP-CTL specific responses to U266, McCAR, MM1S, RPMI, K562 cells or unstimulated cells were analyzed. CD107α upregulation was analyzed by flow cytometry in gated CD8+ T cells. MP-CTLs generated from SMM patients showed elevated levels of CD107α degranulation in response to HLA-A2+ MM cell lines, U266 cell lines, and McCAR cell lines. HLA-A2-restricted degranulation responses against multiple myeloma cells were confirmed using MP-CTLs generated from four SMM patients ( Figure 19a ).

[0526] Example 20: Phenotypic characterization of MP-CTLs generated from relapsed multiple myeloma patients Figure 20a

[0527] The degranulation (CD107α expression) and IFN-γ production of the CD3+CD8+CD137+ subgroup of MP-CTLs generated from smoldering multiple myeloma patients in response to K562-A2 cells presenting individual peptides were analyzed. CD107α and / or IFN-γ expression in the CD3+CD8+

[0528] CD137+ subgroup was analyzed by flow cytometry. MP-CTLs generated from three SMM patients showed elevated levels of peptide-specific IFN-γ production and / or CD107α degranulation ( Figure 20b -d).

[0529] Figure 20c

[0530] The untreated central memory (CM), effector memory (EM), and terminal effector (TE) CD8+ T subgroups of MP-CTLs generated by a total of four multiplex peptide stimulations were analyzed by flow cytometry. MP-CTLs generated from four SMM patients showed increased percentages of EM and TE CD8 + cells ( Figure 20d ). MP-CTLs from SMM patient #2 and SMM patient #4 received an additional three rounds of multiplex peptide stimulation (a total of 7 cycles of stimulation) and were analyzed by flow cytometry. An additional three cycles of MP stimulation led to further expansion of the EM subgroup (cycle 4 vs. cycle 7) ( Example 21: Peptides from unspliced XBP1, spliced XBP1, CD138, and CS1 bind HLA- with high affinity)。In additional experiments, IFN-γ production and CD107α upregulation were evaluated in cancer cell lines with different responses of EM and TE subsets of MP-CTLs generated from three SMM patients. Compared to TE, EM cells in MP-CTLs from all three patients showed a higher percentage of IFN-γ+CD107α+ double-positive cells in response to the HLA-A2+ MM cell line McCAR( Figure 21 ). Additionally, compared to untreated (CD45RO - CCR7 + ) CD8 + T cells, the memory (CD45RO + ) type of CD8 + T cells showed higher expression of the CD69 activation marker in MP-CTLs generated from four smoldering multiple myeloma patients against the HLA-A2 + multiple myeloma cell line U266, confirming the higher anti-tumor activity of the memory subset of MP-CTLs( Figure 22-24 ).

[0531] Example 22: Percentage of CD8+ CTLs in T cells from two donors stimulated with XBP1, CD138, and CS1 peptides A24

[0532] The full-length sequences of unspliced 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, Heidelberg) to predict peptide specificity for HLA-A24, and then the BIMAS program was used to select peptides with an extended half-life dissociation rate. The following peptides from unspliced XBP1 were selected as possible 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 possible 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 possible 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 possible 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 the HIV envelope protein 583-591 (SEQ ID NO:537) (which is known as an HLA-A24-binding peptide) was evaluated using the T2 peptide-binding assay at a peptide concentration of 1 mg / ml. The specific affinity of the peptide was assessed by HLA-A24 mean fluorescence intensity (MFI), which is a function of the upregulation of HLA-A24 on T2 cells after peptide binding to HLA-A24. Among the peptides tested, the following peptides exhibited binding affinity for HLA-A24 equal to or higher than that of the HIV envelope protein 583-591 (SEQ ID NO...

Claims

1. A composition, which comprises: an unspliced XBP1 peptide consisting of the amino acid sequence of SEQ ID NO: 29, a spliced XBP1 peptide consisting of the amino acid sequence of SEQ ID NO: 30, a CD138 peptide consisting of the amino acid sequence of SEQ ID NO: 31, and a CS-1 peptide consisting of the amino acid sequence of SEQ ID NO:

32.

2. A pharmaceutical composition comprising the composition according to claim 1 and a pharmaceutically acceptable carrier.

3. Use of the composition according to claim 1 in the manufacture of a medicament for inducing an immune response in a subject suffering from, at risk of suffering from or in remission from multiple myeloma, breast cancer, colon cancer, pancreatic cancer, prostate cancer, monoclonal gammopathy of undetermined significance (MGUS) or leukemia.

4. Use of the composition according to claim 1 in the manufacture of a medicament for treating a subject suffering from multiple myeloma, breast cancer, colon cancer, pancreatic cancer, prostate cancer, monoclonal gammopathy of undetermined significance (MGUS) or leukemia.

5. Use of the composition according to claim 1 in the manufacture of a medicament for inducing an immune response in a subject suffering from, at risk of suffering from or in remission from smoldering multiple myeloma or for treating a subject suffering from smoldering multiple myeloma.

6. Use according to claim 3 or 4, wherein the breast cancer is estrogen receptor-positive breast cancer, estrogen receptor-negative breast cancer, HER-2-positive breast cancer, HER-2-negative breast cancer, triple-negative breast cancer or inflammatory breast cancer.

7. Use according to any one of claims 3-5, wherein the composition is formulated for administration in combination with one or more additional therapies selected from chemotherapeutic agents, ionizing radiation or immunotherapeutic agents, wherein optionally, the chemotherapeutic agent is selected from capecitabine, cyclophosphamide, gemcitabine or platinum-based agents.

8. Use according to any one of claims 3-5, wherein the composition is formulated for administration in combination with one or more immunostimulants and / or one or more immunomodulators.

9. Use according to claim 8, wherein the one or more immunostimulants are selected from adjuvants including carboxymethylcellulose, polyinosinic acid-polycytidylic acid and poly-L-lysine double-stranded RNA; adjuvants including water and oil emulsions; and adjuvants including proteins.

10. Use according to claim 8, wherein the one or more immunomodulators are selected from antibodies and small molecule adjuvants that activate the immune system.

11. Use according to claim 9, wherein the adjuvant including carboxymethylcellulose, polyinosinic acid-polycytidylic acid and poly-L-lysine double-stranded RNA is poly ICLC.

12. Use according to claim 9, wherein the adjuvant including water and oil emulsions is montanide.

13. Use according to claim 10, wherein the small molecule adjuvant is lenalidomide.

14. The use according to claim 9, in combination with an anti-PD-1 antibody, an anti-PDL-1 antibody, an HDAC inhibitor, or other immunogenic peptides, wherein optionally said other immunogenic peptides are from WT1, MUC1, NY-ESO1 or HER-2.

15. The use according to any one of claims 3-5, wherein the subject expresses HLA-A24.

16. A method of producing the composition of claim 1, said method comprising the steps of: (i) providing a cell comprising a nucleic acid vector comprising a nucleic acid sequence encoding the amino acid sequence of the peptide of claim 1, wherein said nucleic acid sequence is operably linked to an expression control sequence; (ii) culturing the cell under conditions permitting expression of the peptide; and (iii) isolating the peptide from the cell or from the culture medium of the cultured cell.

17. Use of the composition of claim 1 in the manufacture of a medicament for treating and / or inducing an immune response in a subject suffering from, at risk of suffering from, or in remission from: multiple myeloma, breast cancer, colon cancer, pancreatic cancer, prostate cancer, Waldenstrom's macroglobulinemia, monoclonal gammopathy of undetermined significance (MGUS) or leukemia.

18. Use of the composition in the manufacture of a medicament for treating one or more cancer cells in a mammal expressing XBP1, CD138 and / or CS-1, said cancer cells being breast cancer cells, colon cancer cells, pancreatic cancer cells, prostate cancer cells, multiple myeloma cells, Waldenstrom's macroglobulinemia cells, monoclonal gammopathy of undetermined significance (MGUS) cells or blood cells; wherein the composition comprises: (a) an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 29; (b) an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 30; (c) an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 31; and (d) an isolated peptide consisting of the amino acid sequence of SEQ ID NO:

32.

19. The use according to claim 18, wherein the cancer cells are smoldering multiple myeloma cells.

20. Use of the composition in the manufacture of a medicament for treating and / or inducing an immune response in a subject having cancer, at risk of having cancer, or in remission from cancer, said composition comprising: a) an anti-CTLA4 antibody; and b) an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 29; an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 30; an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 31; and an isolated peptide consisting of the amino acid sequence of SEQ ID NO:

32.

21. Use of a composition in the preparation of a medicament for treating and / or inducing an immune response in a subject, wherein the subject has cancer, is at risk of developing cancer, or is in cancer remission, and the composition comprises: a) capecitabine, gemcitabine or oxaliplatin; and b) an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 29; an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 30; an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 31; and an isolated peptide consisting of the amino acid sequence of SEQ ID NO:

32.

22. An ex vivo method of activating multiple T cells, comprising: (i) contacting multiple antigen-presenting cells (APCs) comprising MHC molecules with a composition comprising: (a) an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 29; (b) an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 30; (c) an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 31; and (d) an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 32; (ii) contacting the multiple T cells with the APCs, thereby activating the multiple T cells.

23. Use of multiple T cells produced by the method of claim 22 in the preparation of a medicament for treating and / or inducing an immune response in a subject, wherein the subject has cancer, is at risk of developing cancer, or is in cancer remission.

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