Methods and compositions for cancer treatment using recombinant polypeptides

Abstract

This disclosure provides a method for treating cancer in human subjects, comprising the administration of a recombinant polypeptide containing a cancer-specific CD8+ T cell epitope. The peptide, recognized by a major histocompatibility complex (MHC) molecule, can activate a T cell immune response to target cancer cells in the subject. This disclosure further provides a cancer-specific CD8+ T cell epitope constrained to an MHC molecule expressed by a specific HLA allele in the subject. This disclosure further provides a composition encoding a recombinant polypeptide capable of inducing an augmented memory CD8+ T cell response.

Description

[Technical Field]

[0001] Cross-references to related applications [1] This application claims priority to U.S. Provisional Patent Application No. 63 / 457,918, filed on April 7, 2023, the entirety of which is incorporated herein by reference. [Background technology]

[0002] [2] Esophageal cancer is one of the deadliest cancers in the world and is currently ranked sixth in cancer mortality. Esophageal cancers, including esophageal squamous cell carcinoma and esophageal adenocarcinoma, involve the abnormal proliferation of esophageal epithelial cells. [Overview of the Initiative] [Problems that the invention aims to solve]

[0003] [3] Radiotherapy, surgical resection, and chemotherapy are the primary clinical treatments for esophageal cancer. Outcomes are not optimal, with only 20% of patients surviving for at least 5 years after diagnosis. Survival rates remain low for patients treated with chemotherapy alone, highlighting a significant need for novel targeted therapies that demonstrate long-lasting efficacy. [Means for solving the problem]

[0004] [4] In some embodiments, methods and compositions for treating cancer in a subject, for example, a human subject, using a composition comprising an epitope, for example, a T cell epitope, for example, a cancer-specific CD8+ T cell epitope, are disclosed herein.

[0005] [5] In one embodiment, the present disclosure is a method for treating cancer in a subject requiring treatment for cancer, comprising the step of administering to a subject a composition comprising: (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (ii) a polynucleotide encoding the recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (iii) a vector comprising (i) or (ii); (iv) antigen-presenting cells (APCs) comprising (i), (ii), or (iii); or (v) T cells stimulated by APCs comprising (i) or (ii); wherein the subject is HLA-A * 03:01 MHC expressed by an allele, and the cancer-specific CD8+ T cell epitope is LKPDHIQR, KTEVHGRLK, RVSLPKLGYK, or SLFGARPGR; the subject is HLA-B * NLRPPTQEL is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 07:02 allele; the target is HLA-A * The MHC expressed by the 31:01 allele, and the cancer-specific CD8+ T cell epitope is SLFGARPGR; and / or the subject is HLA-B * This invention provides a method in which LRPPTQEL is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 07:02 allele.

[0006] [6] In one embodiment, the present disclosure is a method for treating cancer in a subject requiring treatment for cancer, comprising the step of administering to a subject a composition comprising: (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (ii) a polynucleotide encoding the recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (iii) a vector comprising (i) or (ii); (iv) antigen-presenting cells (APCs) comprising (i), (ii), or (iii); or (v) T cells stimulated by APCs comprising (i) or (ii); wherein the subject is HLA-A *02:01 Expresses MHC encoded by an allele, and the cancer-specific CD8+ T cell epitope is GVYDGREHTV or KLVELEHTL; the subject is HLA-B * 07:02 Expresses MHC encoded by an allele, and the cancer-specific CD8+ T cell epitope is SPSSASLAL or KPRPDVTNEL; the subject is HLA-B * 35:03 Expresses MHC encoded by an allele, and the cancer-specific CD8+ T cell epitope is SPSSASLAL or KPRPDVTNEL; the subject is HLA-B * 40:01 Expresses MHC encoded by an allele, and the cancer-specific CD8+ T cell epitope is KEFAFLEHSL or HELGFKVVL; the subject is HLA-B * 40:02 Expresses MHC encoded by an allele, and the cancer-specific CD8+ T cell epitope is KEFAFLEHSL or HELGFKVVL; the subject is HLA-B * 44:02 Expresses MHC encoded by an allele, and the cancer-specific CD8+ T cell epitope is KEFAFLEHSL or HELGFKVVL; and / or the subject is HLA-B * 18:01 Provides a method of expressing MHC encoded by an allele, and the cancer-specific CD8+ T cell epitope is HELGFKVVL.

[0007] [7] In one aspect, the present disclosure provides a method of treating cancer in a subject who needs treatment for cancer, comprising administering to the subject a composition comprising: (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (iii) a vector comprising (i) or (ii); (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii); or (v) T cells stimulated with an APC comprising (i) or (ii); wherein the subject is HLA-A *03:01 MHC expressed by an allele, and the cancer-specific CD8+ T cell epitope is SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, or RVFTSSLKTK; the subject is HLA-B * The MHC expressed by the 07:02 allele, and the cancer-specific CD8+ T cell epitope is RPASPRPAP; and / or the subject is HLA-C * This method provides a method in which IATKIALQM is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 03:04 allele.

[0008] [8] In one embodiment, the present disclosure is a method for treating cancer in a subject requiring treatment for cancer, comprising the step of administering to a subject a composition comprising: (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (ii) a polynucleotide encoding the recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (iii) a vector comprising (i) or (ii); (iv) antigen-presenting cells (APCs) comprising (i), (ii), or (iii); or (v) T cells stimulated by APCs comprising (i) or (ii); wherein the subject is HLA-A * 02:01 MHC expressed by the allele, and the cancer-specific CD8+ T cell epitope is GVYDGREHTV or KLVELEHTL; the subject is HLA-A * 03:01 MHC expressed by an allele, and the cancer-specific CD8+ T cell epitope is LKPDHIQR, RVSLPKLGYK, SLFGARPGR, or KTEVHGRLK; the subject is HLA-A * SLFGARPGR is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 31:01 allele; the target is HLA-B *The cells express MHC encoded by the 07:02 allele, and the cancer-specific CD8+ T cell epitope is SPSSASLAL, RPASPRPAP, NLRPPTQEL, or KPRPDVTNEL; the target cells are HLA-B * The cells express MHC encoded by the 35:03 allele, and the cancer-specific CD8+ T cell epitope is SPSSASLAL, RPASPRPAP, NLRPPTQEL, or KPRPDVTNEL; the target cells are HLA-B * The subject expresses MHC encoded by the 40:01 allele, and the cancer-specific CD8+ T cell epitope is either KEFAFLEHSL or HELGFKVVL; the subject is HLA-B * The subject expresses MHC encoded by the 40:02 allele, and the cancer-specific CD8+ T cell epitope is either KEFAFLEHSL or HELGFKVVL; the subject is HLA-B * The subject expresses MHC encoded by the 44:02 allele, and the cancer-specific CD8+ T cell epitope is HELGFKVVL or KEFAFLEHSL; the subject is HLA-B * The MHC expressed by the 18:01 allele, and the cancer-specific CD8+ T cell epitope is HELGFKVVL; and / or the subject is HLA-C * This invention provides a method for expressing MHC encoded by the 07:02 allele, wherein the cancer-specific CD8+ T cell epitope is LRPPTQEL, NLRPPTQEL, or KPRPDVTNEL.

[0009] [9] In one embodiment, the present disclosure is a method for treating cancer in a subject requiring treatment for cancer, comprising the step of administering to a subject a composition comprising: (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (ii) a polynucleotide encoding the recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (iii) a vector comprising (i) or (ii); (iv) antigen-presenting cells (APCs) comprising (i), (ii), or (iii); or (v) T cells stimulated by APCs comprising (i) or (ii); wherein the subject is HLA-A * The MHC expressed by the 15:01 allele, and the cancer-specific CD8+ T cell epitope is GQHLHLETF; the target is HLA-C * IATKIALQM is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 03:04 allele; the target is HLA-C * The subject expresses MHC encoded by the 07:02 allele, and the cancer-specific CD8+ T cell epitope is LRPPTQEL, NLRPPTQEL, or KPRPDVTNEL; the subject is HLA-A * 03:01 cells express MHC encoded by an allele, and the cancer-specific CD8+ T cell epitope is LKPDHIQR, RVSLPKLGYK, SLFGARPGR, KTEVHGRLK, RLHSFTLRQK, RVFTSSLKTK, RVLAKGLAK, KTPLHTLLK, KSYSKVLVR, MTYKIHFKK, or SLYQTIRLK; the subject is HLA-B * The cells express MHC encoded by the 07:02 allele, and the cancer-specific CD8+ T cell epitope is SPSSASLAL, KPRPDVTNEL, NLRPPTQEL, RPASPRPAP, GPRPSPTRSV, SPRSPSPSL, VPQEAVRAPL, VPASPALSR, SPSARPPSL, SPTLNVSAL, or SPSSASLTL; the subject is HLA-B *The subject expresses MHC encoded by the 08:01 allele, and the cancer-specific CD8+ T cell epitope is LNKVKTSL or NLKTEILRL; the subject is HLA-C * The MHC expressed by the 07:01 allele, and the cancer-specific CD8+ T cell epitope is FRGVFVHRY; the target is HLA-B * The MHC expressed by the 35:01 allele, and the cancer-specific CD8+ T cell epitope is TSGPVTEKY; the target is HLA-A * 02:05 The MHC encoded by the allele and the cancer-specific CD8+ T cell epitope is VVAAHLAGA; the target is HLA-A * 02:01 MHC expressed by the allele, and the cancer-specific CD8+ T cell epitope is GVYDGREHTV or KLVELEHTL; the subject is HLA-B * The subject expresses MHC encoded by the 40:01 allele, and the cancer-specific CD8+ T cell epitope is either KEFAFLEHSL or HELGFKVVL; the subject is HLA-B * The subject expresses MHC encoded by the 44:02 allele, and the cancer-specific CD8+ T cell epitope is either KEFAFLEHSL or HELGFKVVL; the subject is HLA-B * The MHC expressed by the 18:01 allele, and the cancer-specific CD8+ T cell epitope is HELGFKVVL; the target is HLA-A * This method provides a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 31:01 allele, and in which case SLFGARPGR is the target of the invention.

[0010]

[10] In one embodiment, the present disclosure provides a method for treating cancer in a subject requiring treatment for cancer, comprising the step of administering to the subject a composition comprising: (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope; (iii) a vector comprising (i) or (ii); (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii); or (v) T cells stimulated by an APC comprising (i), (ii), or (iii).

[0011]

[11] In some embodiments, the subject is HLA-A * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 03:01 allele are LKPDHIQR, KTEVHGRLK, RVSLPKLGYK, or SLFGARPGR.

[0012]

[12] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 07:02 allele is NLRPPTQEL.

[0013]

[13] In some embodiments, the subject is HLA-A * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 31:01 allele is SLFGARPGR.

[0014]

[14] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 07:02 allele is LRPPTQEL.

[0015]

[15] In some embodiments, the subject is HLA-A *The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 02:01 allele are GVYDGREHTV or KLVELEHTL.

[0016]

[16] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 07:02 allele are SPSSASLAL or KPRPDVTNEL.

[0017]

[17] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 35:03 allele are PSSASLAL, RPASPRPAP, NLRPPTQEL, or KPRPDVTNEL.

[0018]

[18] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 35:03 allele are SPSSASLAL or KPRPDVTNEL.

[0019]

[19] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 40:01 allele are KEFAFLEHSL or HELGFKVVL.

[0020]

[20] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 40:02 allele are KEFAFLEHSL or HELGFKVVL. In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 44:02 allele are KEFAFLEHSL or HELGFKVVL.

[0021]

[21] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 18:01 allele is HELGFKVVL.

[0022]

[22] In some embodiments, the subject is HLA-A * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 03:01 allele are SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, or RVFTSSLKTK.

[0023]

[23] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 07:02 allele is RPASPRPAP.

[0024]

[24] In some embodiments, the subject is HLA-C * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 03:04 allele is IATKIALQM.

[0025]

[25] In some embodiments, the subject is HLA-A * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 03:01 allele are LKPDHIQR, RVSLPKLGYK, SLFGARPGR, KSYSKVLVR, MTYKIHFKK, or KTEVHGRLK.

[0026]

[26] In some embodiments, the subject is HLA-A * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 31:01 allele is SLFGARPGR.

[0027]

[27] In some embodiments, the subject is HLA-B *The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 07:02 allele are SPSSASLAL, RPASPRPAP, NLRPPTQEL, VPASPALSR, SPSARPPSL, SPTLNVSAL, or KPRPDVTNEL.

[0028]

[28] In some embodiments, the subject is HLA-C * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 07:02 allele is LRPPTQEL.

[0029]

[29] In some embodiments, the subject is HLA-C * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 07:02 allele are KPRPDVTNEL or NLRPPTQEL.

[0030]

[30] In some embodiments, the subject is HLA-C * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 03:03 allele is NLRPPTQEL.

[0031]

[31] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 15:01 allele is GQHLHLETF.

[0032]

[32] In some embodiments, the subject is HLA-C * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 03:04 allele is IATKIALQM.

[0033]

[33] In some embodiments, the subject is HLA-C * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 07:02 allele is LRPPTQEL.

[0034]

[34] In some embodiments, the subject is HLA-A * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 03:01 allele are LKPDHIQR, RVSLPKLGYK, SLFGARPGR, KTEVHGRLK, RLHSFTLRQK, RVFTSSLKTK, RVLAKGLAK, KTPLHTLLK, KSYSKVLVR, MTYKIHFKK, or SLYQTIRLK.

[0035]

[35] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 07:02 allele are SPSSASLAL, KPRPDVTNEL, NLRPPTQEL, RPASPRPAP, GPRPSPTRSV, SPRPSPSL, VPQEAVRAPL, VPASPALSR, SPSARPPSL, SPTLNVSAL, or SPSSASLTL.

[0036]

[36] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitopes that express MHC encoded by the 08:01 allele are LNKVKTSL or NLKTHLRL.

[0037]

[37] In some embodiments, the subject is HLA-C * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 07:01 allele is FRGVFVHRY.

[0038]

[38] In some embodiments, the subject is HLA-B * The cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 35:01 allele is TSGPVTEKY.

[0039]

[39] In some embodiments, the subject is HLA-A *02:05 The cancer-specific CD8+ T cell epitope that expresses MHC encoded by this allele is VVAAHLAGA.

[0040]

[40] In some embodiments, cancer may be a selection of cancers from colorectal cancer, prostate cancer, esophageal cancer, liver cancer, kidney cancer, lung cancer, bladder cancer, breast cancer, pancreatic cancer, brain cancer, hepatocellular carcinoma, lymphoma, leukemia, stomach cancer, cervical cancer, ovarian cancer, thyroid cancer, melanoma, carcinoma, head and neck cancer, skin cancer, nasopharyngeal cancer, Epstein-Barr-driven cancer, human papillomavirus-driven cancer and soft tissue sarcoma.

[0041]

[41] In some embodiments, polynucleotides may be vectors.

[42] In one embodiment, the Disclosure relates to a composition comprising a recombinant polypeptide or a polynucleotide encoding a recombinant polypeptide, wherein the recombinant polypeptide is, for example, GVYDGREHTV, KLVELEHTL, LKPDHIQR, RVSLPKLGYK, SLFGARPGR, KTEVHGRLK, SLFGARPGR, SPSSASLAL, RPASPRPAP, NLRPPTQEL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, LRPPTQEL, IATKIALQM, SLY The present invention provides a composition comprising cancer-specific CD8+ T cell epitopes, which may include QTIRLK, KTPLHTLLK, RLHSFTLRQK, RVFTSSLKTK, RVLAKGLAK, GPRPSPTRSV, GQHLHLETF, SPRSPSPSL, VPQEAVRAPL, SPSSASLTL, LNKVKTSL, NLKTHLRL, FRGVFVHRY, TSGPVTEKY, KSYSKVLVR, MTYKIHFKK, VPASPALSR, SPSARPPSL, SPTLNVSAL, and / or VVAAHLAGA, or any combination thereof.

[0042]

[43] In one embodiment, the Disclosure provides a composition comprising a recombinant polypeptide or a polynucleotide encoding a recombinant polypeptide, wherein the recombinant polypeptide encoded by the recombinant polypeptide or polynucleotide comprises a cancer-specific CD8+ T cell epitope which may include GVYDGREHTV, KLVELEHTL, LKPDHIQR, RVSLPKLGYK, SLFGARPGR, KTEVHGRLK, SLFGARPGR, SPSSASLAL, RPASPRPAP, NLRPPTQEL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, and / or LRPPTQEL, or any combination thereof.

[0043]

[44] In some embodiments, the composition comprises a recombinant polypeptide or a recombinant polypeptide encoded by a polynucleotide comprising a single cancer-specific CD8+ T cell epitope, which may include, for example, GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL and / or HELGFKVVL, or any combination thereof.

[0044]

[45] In some embodiments, the composition comprises a recombinant polypeptide or a recombinant polypeptide encoded by a polynucleotide comprising a single cancer-specific CD8+ T cell epitope, which may include, for example, LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, and / or LRPPTQEL, or any combination thereof.

[0045]

[46] In some embodiments, the composition comprises a recombinant polypeptide or a recombinant polypeptide encoded by a polynucleotide comprising a single cancer-specific CD8+ T cell epitope, which may include, for example, RPASPRPAP, IATKIALQM, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, and / or RVFTSSLKTK, or any combination thereof.

[0046]

[47] In some embodiments, the recombinant polypeptide or the recombinant polypeptide encoded by a polynucleotide comprises a single cancer-specific CD8+ T cell epitope which may include, for example, IATKIALQM, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, RVFTSSLKTK, RVLAKGLAK, GPRPSPTRSV, GQHLHLETF, SPRSPSPSL, VPQEAVRAPL, SPSSASLTL, LNKVKTSL, NLKTHLRL, FRGVFVHRY, TSGPVTEKY, and / or VVAAHLAGA, or any combination thereof.

[0047]

[48] ​​In one embodiment, the Disclosure relates to a composition comprising a recombinant polypeptide or a vector encoding a polynucleotide encoding a recombinant polypeptide, wherein the recombinant polypeptide is, for example, GVYDGREHTV, KLVELEHTL, LKPDHIQR, RVSLPKLGYK, SLFGARPGR, KTEVHGRLK, SLFGARPGR, SPSSASLAL, RPASPRPAP, NLRPPTQEL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, LRPPTQEL, IATKIALQ The present invention provides a composition comprising cancer-specific CD8+ T cell epitopes, which may include M, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, RVFTSSLKTK, RVLAKGLAK, GPRPSPTRSV, GQHLHLETF, SPRSPSPSL, VPQEAVRAPL, SPSSASLTL, LNKVKTSL, NLKTHLRL, FRGVFVHRY, TSGPVTEKY, KSYSKVLVR, MTYKIHFKK, VPASPALSR, SPSARPPSL, SPTLNVSAL, and / or VVAAHLAGA, or any combination thereof.

[0048]

[49] In some embodiments, the vector encodes a single cancer-specific CD8+ T cell epitope, which may include, for example, GVYDGREHTV, KLVELEHTL, LKPDHIQR, RVSLPKLGYK, SLFGARPGR, KTEVHGRLK, SLFGARPGR, SPSSASLAL, RPASPRPAP, NLRPPTQEL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, and / or LRPPTQEL, or any combination thereof.

[0049]

[50] In some embodiments, the vector encodes a single cancer-specific CD8+ T cell epitope, which may include, for example, GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL and / or HELGFKVVL, or any combination thereof.

[0050]

[51] In some embodiments, the vector encodes a single cancer-specific CD8+ T cell epitope, which may include, for example, LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, and / or LRPPTQEL, or any combination thereof.

[0051]

[52] In some embodiments, the vector encodes a single cancer-specific CD8+ T cell epitope, which may include, for example, RPASPRPAP, IATKIALQM, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, and / or RVFTSSLKTK, or any combination thereof.

[0052]

[53] In some embodiments, the vector encodes a single cancer-specific CD8+ T cell epitope, which may include, for example, IATKIALQM, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, RVFTSSLKTK, RVLAKGLAK, GPRPSPTRSV, GQHLHLETF, SPRSPSPSL, VPQEAVRAPL, SPSSASLTL, LNKVKTSL, NLKTHLRL, FRGVFVHRY, TSGPVTEKY, and / or VVAAHLAGA, or any combination thereof.

[0053]

[54] In some embodiments, the cancer-specific CD8+ T cell epitope may be an esophageal cancer-specific CD8+ T cell epitope.

[55] In some embodiments, the vector may be a vector selected from plasmids, cosmids, bacterial vectors, viral vectors, artificial chromosomes, liposomes, lipid nanoparticles, or exosomes.

[0054]

[56] In some embodiments, the viral vector may be a viral vector, for example, an adenovirus vector, for example, an Ad5 vector, for example, a replication-deficient Adhu5 vector. In some embodiments, the vector is an adenoviral-associated (AAV) vector.

[0055]

[57] In some embodiments, the recombinant polypeptide is 8 to 100 amino acids long. In some embodiments, the recombinant polypeptide may be 8 to 100 amino acids long, for example, 8 to 12 amino acids long, for example, 9 to 10 amino acids long, for example, 9 amino acids long or 10 amino acids long.

[0056]

[58] In some embodiments, cancer-specific CD8+ T cell epitopes can bind to MHC class I molecules with binding affinity of, for example, 500 nM or less.

[59] In some embodiments, cancer-specific CD8+ T cell epitopes can bind to MHC class I molecules with, for example, a predicted binding rank score of 2% or less.

[0057]

[60] In some embodiments, the recombinant polypeptide may contain a single cancer-specific CD8+ T cell epitope.

[61] In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope.

[0058]

[62] In some embodiments, the recombinant polypeptide may contain at least two different cancer-specific CD8+ T cell epitopes.

[63] In some embodiments, the recombinant polypeptide may include at least two different recombinant polypeptides. In some cases, each polypeptide may include a cancer-specific CD8+ T cell epitope.

[0059]

[64] In some embodiments, the composition may further include an adjuvant.

[65] In some embodiments, the method may further include the step of administering a second therapeutic agent, for example, an immune checkpoint inhibitor.

[0060]

[66] In some embodiments, the polynucleotide encoding the recombinant polypeptide may be DNA.

[67] In some embodiments, the polynucleotide encoding the recombinant polypeptide may be RNA, for example, mRNA. In some embodiments, the polynucleotide encoding the recombinant polypeptide is 18 to 45 nucleotides in length.

[0061]

[68] In some embodiments, the composition may be for use in the treatment of cancer, for example, cancer selected from colorectal cancer, prostate cancer, esophageal cancer, liver cancer, kidney cancer, lung cancer, bladder cancer, breast cancer, pancreatic cancer, brain cancer, hepatocellular carcinoma, lymphoma, leukemia, gastric cancer, cervical cancer, ovarian cancer, thyroid cancer, melanoma, carcinoma, head and neck cancer, skin cancer, nasopharyngeal cancer, Epstein-Barr-driven cancer, human papillomavirus-driven cancer and soft tissue sarcoma, for example, esophageal cancer.

[0062]

[69] In some embodiments, the compositions may be for use in the treatment of virus-driven cancers, which may be cancers caused by HPV (human papillomavirus), HTLV (human T-lymphotropic virus), or EBV (Epstein-Barr virus).

[0063]

[70] In some embodiments, the composition can induce an epitope-specific T cell response and / or an augmented memory CD8+ T cell response when administered to a subject.

[0064]

[71] In some embodiments, the composition may be able to induce the production of CD8+ T cells characterized by markers that may include, for example, CX3CR1+, KLRG-1+, CD44+, and / or CD62L-, or any combination thereof. In some embodiments, the composition may be able to induce the production of CD8+ T cells characterized by markers that may include CX3CR1+, KLRG-1+, CD44+, CD62L-, CD27-(low), and / or CD127-(low), or any combination thereof.

[0065]

[72] In some embodiments, the composition may be able to induce the production of CD8+ / CX3CR1+ / KLRG-1+ / CD44+ T cells. In some embodiments, the composition may be able to induce the production of CD8+ / CX3CR1+ / KLRG-1+ / CD62L- T cells. In some embodiments, the composition may be able to induce the production of CD8+ / CX3CR1+ / KLRG-1+ / CD44+ / CD62L- T cells. In some embodiments, the composition may be capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ / CD44+ / CD62L- / CD27- / CD127- T cells, CD8+ / CX3CR1+ / KLRG-1+ / CD44+ / CD62L- / CD27(low) / CD127- T cells, CD8+ / CX3CR1+ / KLRG-1+ / CD44+ / CD62L- / CD27- / CD127(low) T cells, or CD8+ / CX3CR1+ / KLRG-1+ / CD44+ / CD62L- / CD27(low) / CD127(low) T cells.

[0066]

[73] In some embodiments, the composition may be capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ T cells further characterized by the phenotype CCR7- and / or CD45RA+ / -.

[0067]

[74] In some embodiments, the composition may be capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ T cells having sustained expression of Tbx21 and / or E2f2.

[0068]

[75] In some embodiments, the composition may be able to induce the production of CD8+ / CX3CR1+ / KLRG-1+ T cells having low expression of Eomes.

[76] In some embodiments, the composition may be capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ T cells, which may account for about 0.01 percent to 20 percent of the total circulating CD8+ T cells in the subject.

[0069]

[77] In some embodiments, the composition may be capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ T cells that maintain a memory effector phenotype for at least 30 days.

[0070]

[78] In some embodiments, the composition may be capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ T cells having low expression of PD-1, Tim-3, and / or Lag-3.

[0071]

[79] In some embodiments, the composition may be capable of inducing the production of an augmented memory CD8+ T cell response that may be able to control tumor growth in a subject for longer than 50 days after administration of the composition.

[0072]

[80] In some embodiments, the composition may induce the production of a nucleotide sequence encoding a cancer-specific CD8+ T cell epitope, which in some cases is not processed by the antigen-presenting cells of the subject when administered; in some cases, the pharmaceutical composition includes a corresponding viral vector which includes a nucleotide sequence encoding a cancer-specific CD8+ T cell epitope, which in some cases is processed by the antigen-presenting cells of the subject when administered, and the augmented memory CD8+ T cell response is not induced in the subject.

[0073]

[81] In some embodiments, cancer-specific CD8+ T cell epitopes may be viral antigens, tumor-associated antigens overexpressed in cancer cells, or antigens that are mutated in cancer cells.

[0074]

[82] In some embodiments, polypeptides containing cancer-specific CD8+ T cell epitopes cannot be processed by antigen-presenting cells.

[83] In some embodiments, the viral vector may include a CMV promoter and a TATA box.

[0075]

[84] In some embodiments, the viral vector may lack sequences encoding the El and E3 proteins.

[85] In some embodiments, the composition may comprise at least two, three, four, five, or six vectors, each of which is a viral vector, and each of which may encode a different single cancer-specific CD8+ T cell epitope.

[0076]

[86] In some embodiments, the composition may comprise at least one, two, three, four, five, or six vectors, each of which is a viral vector, and each of which may encode two distinct single cancer-specific CD8+ T cell epitopes.

[0077]

[87] In one embodiment, the present disclosure provides pharmaceutical compositions which may comprise the compositions described herein and pharmaceutically acceptable carriers, diluents, excipients, or adjuvants.

[0078]

[88] In some embodiments, the Disclosure provides a method for treating or preventing cancer, which may include the step of administering a therapeutically effective amount of a pharmaceutical composition described herein to a subject in need of treating or preventing cancer.

[0079]

[89] In some embodiments, the present disclosure provides a method for inducing an augmented memory CD8+ T cell response, comprising the step of administering a therapeutically effective amount of a pharmaceutical composition described herein to a subject requiring induction of an augmented memory CD8+ T cell response, wherein the augmented memory CD8+ T cell response may include the production of CD8+ / CX3CR1+ / KLRG-1+ T cells.

[0080]

[90] In some embodiments, the vector or composition may be administered intravenously or intramuscularly.

[91] In some embodiments, the composition may be administered as a single dose.

[0081]

[92] In some embodiments, the composition may be administered as multiple doses, for example, in two doses.

[93] In some embodiments, the first dose is the prime dose.

[0082]

[94] In some embodiments, the first dose induces a measurable immune response in a subject compared to the immune response in the subject in the absence of administration of the first dose.

[95] In some embodiments, the second dose is a booster dose.

[0083]

[96] In some embodiments, the second dose induces a measurable immune response in a subject compared to the immune response in the subject in the absence of administration of the second dose.

[97] In some embodiments, the second dose is administered 24 weeks after the first dose.

[0084]

[98] In some embodiments, the second dose induces an immune response at least twice as high as the immune response induced by the first dose.

[99] In some embodiments, the composition may be administered prophylactically to the subject.

[0085]

[0100] In some embodiments, the composition may be administered in combination with a second therapeutic agent, such as an immune checkpoint inhibitor, a chemotherapeutic agent, a small molecule inhibitor, or a radiotherapy agent.

[0086]

[0101] In some embodiments, the immune checkpoint inhibitor may be an inhibitor of an immune checkpoint protein, such as CTLA-4, PD-1, PD-L1, PD-L2, TIM-3, LAG-3, B7-H3, B7-H4, B7-H6, A2aR, BTLA, GALS and / or IDO, or any combination thereof.

[0087]

[0102] In some embodiments, a method for treating or preventing cancer further includes the step of administering a course of chemotherapy. In some embodiments, a method for treating or preventing cancer further includes the step of administering a first course of chemotherapy and a second course of chemotherapy. In some embodiments, a booster dose is administered two weeks before the second course of chemotherapy.

[0088]

[0103] In some embodiments, the method of treating or preventing cancer further includes surgical procedures, such as surgical resection. In some embodiments, a booster dose is administered one week after recovery following surgery.

[0089]

[0104] In one embodiment, the present disclosure provides a method for producing the compositions described herein, comprising the steps of (i) synthesizing nucleotide sequences encoding a single cancer-specific CD8+ T cell epitope as sense and antisense primers; (ii) cloning the nucleotide sequences encoding the single cancer-specific CD8+ T cell epitope synthesized in (i) into a first plasmid; and (iii) cloning a sequence containing the nucleotide sequence encoding the single cancer-specific CD8+ T cell epitope derived from the first plasmid in (ii) into a second vector containing adenovirus DNA.

[0090]

[0105] In one embodiment, the present disclosure provides a method for treating or preventing cancer, which may include the step of administering a composition described herein to a subject in need of treatment or prevention of cancer, thereby treating cancer in the subject, wherein at least two viral vectors are individually present in the pharmaceutical composition in amounts that are not therapeutically effective.

[0091]

[0106] In some embodiments, the viral vector may include a sequence having at least 90% sequence identity with SEQ ID NO: 44 and a sequence having at least 90% sequence identity with SEQ ID NO: 45.

[0092]

[0107] In some embodiments, the viral vector may further include a sequence having at least 90% sequence identity with SEQ ID NO: 47 and a sequence having at least 90% sequence identity with SEQ ID NO: 48.

[0093]

[0108] In one embodiment, the Disclosure provides a method for treating cancer in a subject requiring treatment for cancer, comprising the steps of: (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, wherein the recombinant polypeptide is bound to the TCR; (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, wherein the recombinant polypeptide encoded by the polynucleotide is bound to the TCR; (iii) a cell comprising the recombinant polypeptide or polynucleotide of (i) or (ii); (iv) a recombinant polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain is bound to a cancer-specific CD8+ T cell epitope; (v) a polynucleotide encoding a recombinant polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain is bound to a cancer-specific CD8+ T cell epitope; or (vi) a cell comprising the recombinant polypeptide or polynucleotide of (iii) or (iv); and thereby treating the subject.

[0094]

[0109] In some embodiments, the viral vector comprises one or more of a promoter, a translation initiation sequence, a start codon, a T cell epitope codon, a stop codon, and a polyadenylation sequence, wherein (i) the promoter is selected from the CMV promoter, the RSV promoter, and the EFiα promoter, and optionally the promoter comprises the sequence shown in SEQ ID NO: 36; (ii) the translation initiation sequence comprises the sequence shown in SEQ ID NO: 37; and (iii) the polyadenylation sequence comprises the sequence shown in SEQ ID NO: 43.

[0095]

[0110] In some embodiments, the vector or viral vector includes a deletion or functional deletion in the E1 gene at the E1 gene locus. In some embodiments, the vector or viral vector includes a transgene insertion at the E1 gene locus.

[0096]

[0111] In some embodiments, the method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence GVYDGREHTV, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KLVELEHTL; the subject is HLA-A * 02:01 Expresses MHC encoded by the allele.

[0097]

[0112] In some embodiments, the method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence SPSSASLAL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KPRPDVTNEL; the subject is HLA-B * 07:02 expresses MHC encoded by the allele.

[0098]

[0113] In some embodiments, the method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence SPSSASLAL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KPRPDVTNEL; the subject is HLA-B * 35:03 expresses MHC encoded by this allele.

[0099]

[0114] In some embodiments, the method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KEFAFLEHSL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence HELGFKVVL; the subject is HLA-B *It expresses MHC encoded by the 40:01 allele.

[0100]

[0115] In some embodiments, the method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KEFAFLEHSL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence HELGFKVVL; the subject is HLA-B * It expresses MHC encoded by the 40:02 allele.

[0101]

[0116] In some embodiments, the method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KEFAFLEHSL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence HELGFKVVL; the subject is HLA-B * It expresses MHC encoded by the 44:02 allele.

[0102]

[0117] In some embodiments, the method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KEFAFLEHSL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence HELGFKVVL; the subject is HLA-B * 18:01 expresses MHC encoded by the allele.

[0103]

[0118] In some embodiments, the composition comprises (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence GVYDGREHTV, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KLVELEHTL.

[0104]

[0119] In some embodiments, the composition is intended for use in the treatment of esophageal cancer in a subject.

[0120] In some embodiments, the composition is intended for use in the treatment of esophageal cancer in subjects, and the subjects are HLA-A * 02:01 Expresses MHC encoded by the allele.

[0105]

[0121] In some embodiments, the composition comprises (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence SPSSASLAL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KPRPDVTNEL.

[0106]

[0122] In some embodiments, the composition is intended for use in the treatment of esophageal cancer in a subject.

[0123] In some embodiments, the composition is intended for use in the treatment of esophageal cancer in subjects, and the subjects are HLA-B * 07:02 expresses MHC encoded by the allele.

[0107]

[0124] In some embodiments, the composition is intended for use in the treatment of esophageal cancer in subjects, and the subjects are HLA-B * 35:03 expresses MHC encoded by this allele.

[0108]

[0125] In some embodiments, the composition comprises (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KEFAFLEHSL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence HELGFKVVL.

[0109]

[0126] In some embodiments, the composition is intended for use in the treatment of esophageal cancer in a subject.

[0127] In some embodiments, the composition is intended for use in the treatment of esophageal cancer in subjects, and the subjects are HLA-B * It expresses MHC encoded by the 40:01 allele.

[0110]

[0128] In some embodiments, the composition is intended for use in the treatment of esophageal cancer in subjects, and the subjects are HLA-B * It expresses MHC encoded by the 40:02 allele.

[0111]

[0129] In some embodiments, the composition is intended for use in the treatment of esophageal cancer in subjects, and the subjects are HLA-B * It expresses MHC encoded by the 44:02 allele.

[0112]

[0130] In some embodiments, the composition is intended for use in the treatment of esophageal cancer in subjects, and the subjects are HLA-B * 18:01 expresses MHC encoded by the allele.

[0113]

[0131] In some embodiments, the first adenovirus vector is an Ad5 adenovirus vector, and / or the second adenovirus vector is an Ad5 adenovirus vector.

[0114]

[0132] In some embodiments, the first adenovirus vector includes a deletion or functional deletion in the E1 gene at the E1 gene locus, and / or the second adenovirus vector includes a deletion or functional deletion in the E1 gene at the E1 gene locus. Built-in by reference

[0133] All publications, patents, and patent applications cited herein are incorporated by reference to the same extent as each individual publication, patent, or patent application is specifically and individually indicated as being incorporated by reference.

[0115]

[0134] The features of this disclosure are described in detail in the appended claims. A better understanding of these features and this disclosure will be obtained from the following detailed description illustrating exemplary embodiments in which the principles of the present invention are utilized, and from reference to the appended drawings (also referred to herein as "Figure" and "FIG."). [Brief explanation of the drawing]

[0116] [Figure 1]

[0135] Figure 1 shows a schematic diagram of the proposed clinical trial protocol. [Figure 2]

[0136] Figure 2 shows a schematic diagram of the proposed clinical trial study design. [Figure 3]

[0137] Figure 3 shows the events at the time points of the proposed clinical trial design. [Modes for carrying out the invention]

[0117]

[0138] Immunotherapy approaches are widely used to treat cancer by modulating T cells to recognize cancer cells, thereby causing white blood cells to attack the cancer. Epitope-based cancer vaccines are one strategy used to activate the T cell response to specific tumor-associated antigens (TAAs). TAAs are highly expressed in tumor cells and to a lower degree in normal tissues. Initially, peptide-based monoepitope vaccines were used, but these provided insufficient clinical responses because they did not adequately activate the innate immune system.

[0118]

[0139] This disclosure provides recombinant polypeptides comprising at least one T cell epitope, e.g., a CD8+ T cell epitope, e.g., a cancer-specific CD8+ T cell epitope; polynucleotides encoding these T cell epitopes, e.g., CD8+ T cell epitopes; vectors; cells; compositions; pharmaceutical compositions; formulations; and methods for using them.

[0119]

[0140] The practices described herein employ conventional techniques of immunology, molecular biology, chemistry, biochemistry, and recombinant DNA technology, which are within the scope of the art, unless otherwise indicated. Such techniques are fully described in the literature, see, for example, Green and Sambrook et al., Molecular Cloning: A Laboratory Manual, 4th edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (2012). definition

[0141] Unless otherwise defined, all terms, notations, and other technical and scientific or specialized terms used herein are intended to have the same meaning as those commonly understood by those skilled in the art in the field relating to the subject matter of the claims. In some cases, terms that have a commonly understood meaning are defined herein for clarity and / or for easy reference, and the inclusion of such definitions herein should not necessarily be construed as representing a substantial difference as commonly understood in the art. Specialized terms used herein are for the purpose of describing a particular embodiment and are not intended to limit it.

[0120]

[0142] As used herein and in the claims, the singular forms "a," "an," and "the" refer to multiple objects unless the context explicitly points to another. For example, the term "a sample" refers to multiple samples, including mixtures thereof.

[0121]

[0143] Furthermore, to that extent, the terms “including,” “includes,” “having,” “has,” and “with,” as used herein, or variations thereof, mean “comprising.” Unless otherwise specified, the terms “comprising,” “comprise,” “comprises,” “having,” “have,” “has,” “including,” “includes,” “include,” “containing,” “contains,” and “contain” are inclusive or open-ended and do not exclude additional unenumerated elements or steps of method.

[0122]

[0144] In this specification, the term "and / or" as used in phrases such as "X and / or Y" is intended to include both X and Y; X or Y; X (alone); and Y (alone).

[0123]

[0145] References to “some embodiments,” “a certain embodiment,” “one embodiment,” or “other embodiments” mean that certain features or characteristics described in relation to an embodiment are included in at least one or more embodiments of the Disclosure, but not necessarily in all embodiments of the Disclosure.

[0124]

[0146] In this specification, the terms “about” or “approximately” refer to a range of values ​​that are greater than or less than 15% of a given value.

[0147] Scope: Throughout this disclosure, various aspects of the disclosure may be represented in scope format. It should be understood that the use of scope format is merely for convenience and brevity and should not be interpreted as a definitive limitation to the scope of the disclosure. Accordingly, scope statements should be considered to have not only all possible sub-ranges specifically disclosed, but also the individual numbers within that range. For example, a scope statement such as 1-6 should be considered to have not only the specifically disclosed sub-ranges, e.g., 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, etc., but also the individual numbers within that range, e.g., 1, 2, 2.7, 3, 4, 5, 5.3, and 6. As another example, a range such as 95-99% identity includes those with 95%, 96%, 97%, 98%, or 99% identity, and also includes subranges such as 96-99%, 96-98%, 96-97%, 97-99%, 97-98%, and 98-99% identity. This applies regardless of the breadth of the range.

[0125]

[0148] As used herein, the terms “determining,” “measuring,” “evaluating,” “assessing,” “assaying,” and “analyzing” are often used interchangeably to refer to forms of measurement. These terms include determining whether an element is present or absent (e.g., detection). These terms may include quantitative, qualitative, or a combination of quantitative and qualitative determination. Assessment may be relative or absolute. “Detecting the presence” may include determining the quantity of something present, in addition to determining whether it is present or absent depending on the context.

[0126]

[0149] As used herein, the term “cancer” refers to a disease characterized by abnormal cell growth, and as used herein, the term refers to both primary tumors and metastases of primary tumors. Cancer cells can spread locally or through the bloodstream and lymphatic system to other parts of the body. Examples of various cancers, but not limited to, include colorectal cancer, prostate cancer, esophageal cancer, liver cancer, kidney cancer, lung cancer, breast cancer, pancreatic cancer, brain cancer, hepatocellular carcinoma, lymphoma, leukemia, stomach cancer, cervical cancer, ovarian cancer, thyroid cancer, melanoma, carcinoma, head and neck cancer, skin cancer, nasopharyngeal cancer, Epstein-Barr-driven cancer, human papillomavirus-driven cancer, and soft tissue sarcoma. Cancers described herein may be stage I, stage II, stage III, or stage IV cancers. As used herein, the term “tumor” refers to a physiological condition in mammals characterized by uncontrolled cell growth.

[0127]

[0150] Where used herein, the terms “individual,” “patient,” or “subject” refer to an individual diagnosed with, suspected of suffering from, or at risk of developing, at least one disease for which the described compositions and methods are useful to treat. In certain embodiments, the individual is a human being.

[0128]

[0151] In some embodiments, the individual is a mammal. In some embodiments, the mammal is a non-human primate (e.g., a rhesus monkey or other type of macaque), mouse, rat, rabbit, dog, cat, horse, cattle, sheep, pig, goat, llama, alpaca, or yak. ​​In certain embodiments, the individual is a human.

[0129]

[0152] As used herein, the term "linker" refers to a molecule that connects two other molecules or parts. A linker may be an amino acid sequence when it joins two fusion proteins. A linker may also be a nucleotide sequence when it joins two nucleotide sequences together. In some cases, a linker may have varying lengths depending on the application of the linker or sequence, or the molecules being linked by the linker.

[0130]

[0153] Where used herein, the terms “treatment” or “treating” are used to refer to a pharmaceutical regimen or other intervention regimen used to obtain beneficial or desired outcomes in a recipient. Beneficial or desired outcomes include, but are not limited to, therapeutic and / or preventive benefits. A therapeutic benefit may refer to the eradication or improvement of the symptom or underlying disorder being treated. A therapeutic benefit may also be achieved by the eradication or improvement of one or more physiological symptoms associated with an underlying disorder, even though the subject may still suffer from the underlying disorder, resulting in an observed improvement in the subject. A preventive effect includes delaying, preventing, or eliminating the onset of a disease or condition; delaying or eliminating the onset of symptoms of a disease or condition; slowing, stopping, or reversing the progression of a disease or condition; or any combination thereof. With respect to preventive benefits, subjects at risk of developing a particular disease, or those reporting one or more physiological symptoms of a disease, may receive treatment even if they have not been diagnosed with the disease. Those skilled in the art will recognize that a given population of individuals potentially for treatment may not respond at all to treatment or may not respond at all to treatment. Such individuals are considered to be treated when administered a composition comprising the pharmaceutical composition described herein.

[0131]

[0154] As used herein, the terms “effective dose,” “therapeutic dose,” or “pharmaceutical dose” refer to an amount sufficient to produce a treatment when administered to a subject (e.g., a mammal such as a human) that requires such treatment, as defined below. The therapeutic dose or pharmaceutical dose varies depending on the subject and disease state being treated, the subject’s weight and age, the severity of the disease state, the mode of administration, etc., which can be readily determined by those skilled in the art. For example, the “therapeutic dose” or “pharmaceutical dose” of AAV2-derived capsid as described herein is an amount sufficient to produce an immune response in the subject (e.g., a human). In some embodiments, the immune response is sufficient to increase the AAV capsid to neutralize antibodies against the relevant capsid in the subject.

[0132]

[0155] As used herein, the term “sequence identity” refers to subunit sequence identity between two polymer molecules, for example, between two polynucleotide or polypeptide sequences. Sequence identity analysis begins by aligning the two sequences. Identical sequences (100% sequence identity) have the same nucleotide or amino acid at each position in the alignment. “Sequence identity percentage” is determined by comparing the number of identical positions in the sequence alignment with the total number of subunits. Sequence identity percentage can be determined over a portion of the sequence or over the entire sequence. Sequence identity percentage can be determined using a sequence comparison algorithm. Test and reference sequences are input into a computer, and, if necessary, subsequence coordinates are designed, and parameters for the sequence algorithm program are designed. The sequence comparison algorithm then aligns the sequences and calculates the sequence identity percentage based on the designed program parameters.

[0133]

[0156] Sequence identity is typically measured using sequence analysis software. Optimal alignment of sequences for comparison can be achieved, for example, by the local homology algorithm of Smith and Waterman, Adv.Appl.Math.2:482 (1981), the homology alignment algorithm of Needleman and Wunsch, J.Mol.Biol.48:443 (1970), the search for similarity method of Pearson and Lipman, Proc.Nat'l.Acad.Sci.USA 85:2444 (1988), computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics software package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (see Ausubel et al., Current Protocols in Molecular Biology). Such software matches identical or similar sequences by assigning degrees of homology to various substitutions, deletions, and / or other modifications. In an exemplary approach to determining the degree of identity, the BLAST program may be used with probability scores from e-3 to e-100 indicating closely related sequences. An example of a suitable algorithm for determining sequence identity and sequence similarity percentages is the BLAST algorithm, described in Altschul et al., J.Mol.Biol.215:403 (1990). Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information (publicly accessible through the National Institutes of Health NCBI Internet Server). Typically, sequence comparisons can be performed using default program parameters, but customized parameters can also be used.For amino acid sequences, the BLASTP program uses a word length (W) of 3, an expected length (E) of 10, and a BLOSUM62 scoring matrix by default (see Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989)). In some embodiments, sequence identity is determined by Needleman-Wunsch alignment of two sequences using gap costs set to presence: 11, extension: 1, where the identity percentage is calculated by dividing the number of identities by the length of the alignment. Conservative substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine.

[0134]

[0157] As used herein, the terms “antigen-presenting cell” or “APC” refer to immune system cells, such as accessory cells (e.g., B cells, dendritic cells, etc.), that display foreign antigens that are complexed with major histocompatibility complexes (MHCs) on their surface. T cells can recognize these complexes using their T cell receptors (TCRs). APCs process antigens and present them to T cells.

[0135]

[0158] The term "encoding" refers to the specific properties of a particular sequence of nucleotides in a gene, cDNA, or mRNA, whether a defined sequence of nucleotides (e.g., rRNA, tRNA, and mRNA) or a defined sequence of amino acids, and the biological properties derived therefrom, which serve as a template for the synthesis of other polymers and macromolecules in biological processes. Thus, a gene, cDNA, or RNA codes for a protein if the transcription and translation of the mRNA corresponding to that gene produces a protein in a cell or other biological system. Both the coding strand, whose nucleotide sequence is identical to the mRNA sequence and is typically provided in a sequence listing, and the non-coding strand, which is used as a template for the transcription of a gene or cDNA, can be said to code for a protein or other product of that gene or cDNA.

[0136]

[0159] Unless otherwise specified, “nucleotide sequences encoding an amino acid sequence” includes all nucleotide sequences that encode the same amino acid sequence, including degenerate versions of each other. The term “nucleotide sequence encoding a protein” or “RNA” may also include introns to the extent that a protein-coding nucleotide sequence may contain one or more introns in some versions.

[0137]

[0160] As used herein, the term "T cell" and its grammatical equivalent may refer to a T cell of any origin. For example, a T cell may be a primary T cell, such as an autologous T cell, an allogeneic T cell, or a cell line. A T cell may also be of human or non-human origin.

[0138]

[0161] As used herein, the term "CD8" refers to the surface antigen classification 8 protein, which is a transmembrane glycoprotein that acts as a co-receptor for the T cell receptor (TCR).

[0139]

[0162] As used herein, the term “effector T cell” includes helper T (e.g., CD4+) cells and cytotoxic (e.g., CD8+) T cells. CD4+ effector T cells contribute to the development of several immunological processes, including the maturation of B cells into plasma cells and memory B cells, as well as the activation of cytotoxic T cells and macrophages. CD8+ effector T cells destroy virus-infected cells and tumor cells.

[0140]

[0163] As used herein, the terms “nucleic acid” or “polynucleotide” refer to deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), and polymers thereof in either single-stranded or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogs of native nucleotides that have similar binding properties to a reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses its conservatively modified variants (e.g., degenerate codon substitutions), alleles, orthologues, SNPs, and complementary sequences, as well as sequences explicitly indicated. Specifically, degenerate codon substitution can be achieved by creating sequences in which the third position of one or more selected (or all) codons is replaced with a mixed base and / or a deoxyinosine residue (Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605~2608 (1985); and Rossolini et al., Mol. Cell. Probes 8:91~98 (1994)).

[0141]

[0164] As used herein, the terms “peptide,” “polypeptide,” and “protein” are interchangeable and refer to compounds composed of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and there is no limit to the maximum number of amino acids that may make up a protein or peptide sequence. A polypeptide includes any peptide or protein containing two or more amino acids linked to each other by peptide bonds. As used herein, this term refers to both short chains, oligopeptides and oligomers, which are also commonly referred to in the art as peptides, and longer chains, of which there are many types, which are commonly referred to in the art as proteins. A “polypeptide” includes, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, polypeptide variants, modified polypeptides, derivatives, analogs, fusion proteins, and the like. Polypeptides include native peptides, recombinant peptides, or combinations thereof.

[0142]

[0165] The term "promoter" refers to a DNA sequence that is recognized by a cell's transcriptional or introduced synthetic mechanism, which is necessary to initiate the specific transcription of a polynucleotide sequence.

[0143]

[0166] The term "promoter / regulatory sequence" refers to a nucleic acid sequence required for the expression of a gene product operably linked to a promoter / regulatory sequence. In some examples, this sequence may be a core promoter sequence, and in other examples, it may also include enhancer sequences and other regulatory elements required for the expression of the gene product. The promoter / regulatory sequence may, for example, express the gene product in a tissue-specific manner.

[0144]

[0167] As used herein, the term “vector” refers to a nucleic acid sequence that can be used to transfer another nucleic acid, to which the vector sequence is ligated, into a cell. A number of vectors are known in the art, but are not limited to linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Examples of viral vectors include, but are not limited to, adenovirus vectors, adeno-associated virus vectors (AAVs), retroviral vectors, and lentiviral vectors. The vectors of this disclosure may be adenoviruses and comprise a nucleotide sequence encoding a single cancer-specific CD8+ T cell epitope containing a gene construct (e.g., ligated to a transcriptional regulatory element) in a form suitable for expression by cells.

[0145]

[0168] As used herein, the term “epitope” refers to a portion of an antigen recognized by the immune system, which may be a short protein sequence. “Cancer-specific CD8+ T cell epitope” refers to an epitope that may be presented by an antigen-presenting cell bound to an MHC molecule and subsequently recognized by a T cell receptor (TCR). CD8+ T cells express a CD8 coreceptor, which binds to MHC I and recognizes peptides presented by the MHC I molecule.

[0146]

[0169] As used herein, the terms "HLA-binding affinity" or "MHC-binding affinity" mean the affinity for binding between a specific antigen and a specific MHC allele.

[0147]

[0170] Where used herein, the terms “HLA-binding rank score” or “MHC-binding rank score” mean predictive rank score. The predictive rank score is the rank position percentage of a query sequence (antigen) relative to the distribution of predictive scores for the MHC of question, estimated from a random set of native peptides. Rank scores below 2% indicate binding between a specific antigen and a specific MHC allele. For example, a method for treating cancer in a human subject is provided herein by administering a recombinant polypeptide (or polynucleotide encoding a recombinant polypeptide) comprising a T-cell epitope capable of binding to one or more MHC molecules, which is presented by one or more MHC molecules and capable of activating T cells and their compositions that are immunogenic and / or cytotoxic. In some embodiments, the epitopes described herein (e.g., a single cancer-specific CD8+ T cell epitope) may bind to MHC (e.g., MHC class I) molecules with rank scores (e.g., binding rank scores) of at least about 0.5%, at least about 1%, at least about 1.5%, at least about 2%, at least about 2.5%, at least about 3%, at least about 3.5%, at least about 4%, at least about 5%, or more than about 5%. In some embodiments, the epitopes described herein (e.g., a single cancer-specific CD8+ T cell epitope) may bind to MHC (e.g., MHC class I) molecules with rank scores (e.g., binding rank scores) of up to about 5%, up to about 4%, up to about 3.5%, up to about 3%, up to about 2.5%, up to about 2%, up to about 1.5%, up to about 1%, up to about 0.5%, or less than about 0.5%. Recombinant polypeptides

[0171] In one embodiment, the disclosure provides a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope. In some embodiments, the single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is an immunogenic epitope, e.g., it induces an immune response. In some embodiments, the single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope binds to the major histocompatibility complex (MHC complex), e.g., MHC associated with cells, e.g., cancer cells. In some embodiments, a single epitope, for example, a single T cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a single cancer-specific CD8+ epitope, can bind to and present MHC molecules, cells, for example, MHC molecules associated with cancer cells.

[0148]

[0172] MHC molecules are highly polymorphic proteins that regulate T cell responses (Schwartz, BD, *The human major histocompatibility complex HLA in basic & clinical immunology*, edited by Stites et al., Lange Medical Publication: Los Altos, pp. 52-64, 4th edition). The MHC system is responsible for presenting epitopes to responding T cells. Species-specific MHC homologs that display cytotoxic T cell lymphocyte (CD8+) epitopes or helper T cell lymphocyte (CD4+) epitopes in humans are called human leukocyte antigens ("HLA"). HLA class I and class II molecules can be divided into several families or "supertypes" based on their ability to bind to a similar repertoire of peptides. HLA class I (HLA-A, -B, and -C) and peptide complexes are recognized by CD8+ T cells, which can then target presenting cells for destruction. HLA class II (HLA-DP, -DQ, and -DR) and peptide complexes are recognized by CD4+ T cells, which can then initiate an immune response against the presenting cells. To trigger an immune response, tumor-specific peptides must bind to MHC molecules. This binding depends on the alleles of the MHC molecule and the specific polymorphism of the amino acid sequence of the epitope. In some cases, recombinant polypeptides can contain cancer-specific CD8+ T cell epitopes that can bind to MHC class I molecules. In some cases, recombinant polypeptides can contain cancer-specific CD4+ T cell epitopes that can bind to MHC class II molecules.

[0149]

[0173] In some embodiments, an epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ epitope, binds to MHC encoded by HLA, e.g., an HLA-A allele, e.g., an HLA-B allele, e.g., an HLA-C allele. In some embodiments, the HLA-A allele binds to HLA-A * 11 alleles (for example, HLA-A* 11:01 allele), HLA-A * 02 allele (e.g., HLA-A * 02:01 allele, e.g., HLA-A * 02:05 allele), HLA-A * 03 allele (e.g., HLA-A * 03:01 allele), or HLA-A * 31 allele (e.g., HLA-A * 31:01 allele). In some embodiments, the HLA-B allele is HLA-B * 07 allele (e.g., HLA-B * 07:02 allele), HLA-B * 08 allele (e.g., HLA-B * 08:01 allele), HLA-B * 15 allele (e.g., HLA-B * 15:01 allele), HLA-B * 18 allele (e.g., HLA-B * 18:01 allele), HLA-B * 35 allele (e.g., HLA-B * 35:01 allele, HLA-B * 35:03 allele), HLA-B * 40 allele (e.g., HLA-B * 40:01 allele, e.g., HLA-B * 40:02 allele), or HLA-B * 44 allele (e.g., HLA-B * 44:01 allele, e.g., HLA-B * 44:02 allele). In some embodiments, the HLA-C allele is HLA-C * 03 allele (e.g., HLA-C * 03:04 allele), or HLA-C * 07 allele (e.g., HLA-C * 07:01 allele, e.g., HLA-C *07:02 allele). In some embodiments, a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, can bind to multiple MHCs encoded by different HLAs, e.g., a single T cell epitope. In some embodiments, the single T cell epitope disclosed herein, e.g., a single cancer-specific CD8+ epitope, is HLA-B * 40:01, HLA-B * 44:01, HLA-B * 44:02, or HLA-B * It can bind to MHC encoded by an HLA selected from 18:01. In some embodiments, the epitopes disclosed herein, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ epitope, bind to MHC encoded by the HLA allele to which it is predicted to bind.

[0150]

[0174] Predicted binding affinity and rank for MHC molecules can be useful in prioritizing peptides for experimental testing. Several methods for predicting peptide binding to MHC class I and MHC class II complexes are known in the art, but are not limited to, NetMHC, NetMHCpan, NetMHCllpan, Stabilization Matrix Method (SMM), and / or Mean Relative Binding Matrix (ARB). In some examples, predicted binding rank scores may be less than 0.1% including 4%–0%, 3.5%–0.2%, 3%–0.5%, 2.5%–1%, 2.25%–1.5%, 2%–1%, 2%–0.5%, 0.5%–0.4%, 0.4%–0.3%, 0.3%–0.2%, 0.2%–0.1%, or less than 0.1% including 0%.

[0151]

[0175] In some examples, the binding affinity may be about 500 nM or less than 500 nM. For example, the binding affinity may be less than about 100 nM, including about 500 nM to 400 nM, about 400 nM to 300 nM, about 300 nM to 200 nM, about 200 nM to 100 nM, or 0 nM. In some embodiments, the epitopes described herein (e.g., a single cancer-specific CD8+ T cell epitope) may bind to MHC (e.g., MHC class I) molecules with binding affinities of up to about 750 nM, up to about 500 nM, up to about 400 nM, up to about 300 nM, up to about 200 nM, up to about 100 nM, up to about 50 nM, or less than about 50 nM.

[0152]

[0176] In some embodiments, binding of a single epitope to an MHC complex, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, initiates a subsequent immune response, e.g., an augmented memory T cell response, e.g., an augmented memory CD8+ and / or CD4+ T cell response, e.g., an augmented memory CD8 epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single T cell epitope, e.g., a In some embodiments, recombinant polypeptides comprising a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or epitope, e.g., a single cancer-specific CD8+ epitope, do not bind to the HLA type encoding the MHC of the patient's non-cancer cells. In some embodiments, recombinant polypeptides comprising a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, do not cause T-cell exhaustion while binding to the HLA type encoding the MHC of the patient's cancer cells.

[0153]

[0177] There are several methods known in the art for identifying epitopes that bind to MHC and thus produce an immune response, such as peptide-MHC binding predictive models, of which there are several publicly available programs.

[0154]

[0178] In some embodiments, the recombinant polypeptide comprises a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence selected from Table 1. In some embodiments, the recombinant polypeptide comprises a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence having 90-99% identity with the amino acid sequence shown in any of SEQ ID NOs: 1-35. In some embodiments, the recombinant polypeptide comprises a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence having 90-99% identity with the amino acid sequence shown in any of SEQ ID NOs: 1-35. In some embodiments, the recombinant polypeptide comprises a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific

[0155]

[0179] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence shown in any of SEQ ID NOs: 1 to 13. In some embodiments, the recombinant polypeptide comprises a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence having 90 to 99% identity with the amino acid sequence shown in any of SEQ ID NOs: 1 to 13. In some embodiments, the recombinant polypeptide comprises a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence having one, two, or up to three amino acid mutations compared to the amino acid sequence shown in any of SEQ ID NOs: 1 to 13.

[0156]

[0180] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence shown in any of SEQ ID NOs: 1-6. In some embodiments, the recombinant polypeptide comprises a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence having 90-99% identity with the amino acid sequence shown in any of SEQ ID NOs: 1-6. In some embodiments, the recombinant polypeptide comprises a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence having one, two, or up to three amino acid mutations compared to the amino acid sequence shown in any of SEQ ID NOs: 1-6.

[0157]

[0181] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence shown in any of SEQ ID NOs. 7 to 12. In some embodiments, the recombinant polypeptide comprises a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence having 90-99% identity with the amino acid sequence shown in any of SEQ ID NOs. 7 to 12. In some embodiments, the recombinant polypeptide comprises a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence having one, two, or up to three amino acid mutations compared to the amino acid sequence shown in any of SEQ ID NOs. 7 to 12.

[0158]

[0182] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence shown in any of SEQ ID NOs. 13-18. In some embodiments, a recombinant polypeptide comprises a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence having 90-99% identity with the amino acid sequence shown in any of SEQ ID NOs. 13-18. In some embodiments, a recombinant polypeptide comprises a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence having one, two, or up to three amino acid mutations compared to the amino acid sequence shown in any of SEQ ID NOs. 13-18. In some embodiments, a single epitope, for example, a single T-cell epitope, for example, a single cancer-specific CD8+ epitope, comprises an amino acid sequence selected from any of SEQ ID NOs: 19-35, an amino acid sequence having 90-99% identity with an amino acid sequence shown in any of SEQ ID NOs: 19-35, or an amino acid sequence having one, two, or up to three amino acid mutations compared to an amino acid sequence shown in any of SEQ ID NOs: 19-35.

[0159] [Table 1-1]

[0160] [Table 1-2]

[0161]

[0183] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises an amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises an amino acid sequence having 90 to 99% identity with the amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises an amino acid sequence with one, two, or up to three amino acid mutations compared to the amino acid sequence shown in any of SEQ ID NOs: 1 to 35.

[0162]

[0184] In some embodiments, a single cancer-specific T cell epitope (e.g., a single cancer-specific CD8+ T cell epitope) may contain an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5%, at least about 99.9%, or more than about 99.9% sequence identity with the amino acid sequence shown in any one of SEQ ID NOs: 1 to 35. In some embodiments, a single cancer-specific T cell epitope (e.g., a single cancer-specific CD8+ T cell epitope) may contain an amino acid sequence having up to about 99.9%, up to about 99.5%, up to about 99%, up to about 98.5%, up to about 98%, up to about 97%, up to about 96%, up to about 95%, up to about 94%, up to about 93%, up to about 92%, up to about 91%, up to about 90%, or less than about 90% sequence identity with the amino acid sequence shown in any one of SEQ ID NOs: 1 to 35.

[0163]

[0185] In some embodiments, a single cancer-specific T cell epitope (e.g., a single cancer-specific CD8+ T cell epitope) may contain an amino acid sequence having about 90% to about 99.9%. T cell epitopes are approximately 90% to 91%, 90% to 92%, 90% to 93%, 90% to 94%, 90% to 95%, 90% to 96%, 90% to 97%, 90% to 98%, 90% to 99%, 90% to 99.5%, 90% to 99.9%, 91% to 92%, 91% to 93%, 91% to 94%, 91% to 95%, 91% to 96%, and 91% to 97%. Approximately 91% to 98%, approximately 91% to 99%, approximately 91% to 99.5%, approximately 91% to 99.9%, approximately 92% to 93%, approximately 92% to 94%, approximately 92% to 95%, approximately 92% to 96%, approximately 92% to 97%, approximately 92% to 98%, approximately 92% to 99%, approximately 92% to 99.5%, approximately 92% to 99.9%, approximately 93% to 94%, approximately 93% to 95%, approximately 93% to 96%, approximately 93% to 97%, approximately 93% to Approximately 98%, approximately 93% to 99%, approximately 93% to 99.5%, approximately 93% to 99.9%, approximately 94% to 95%, approximately 94% to 96%, approximately 94% to 97%, approximately 94% to 98%, approximately 94% to 99%, approximately 94% to 99.5%, approximately 94% to 99.9%, approximately 95% to 96%, approximately 95% to 97%, approximately 95% to 98%, approximately 95% to 99%, approximately 95% to 99.5%, approximately 95% to 99.9%, approximately 96% to 9 It may contain amino acid sequences having 7%, approximately 96% to approximately 98%, approximately 96% to approximately 99%, approximately 96% to approximately 99.5%, approximately 96% to approximately 99.9%, approximately 97% to approximately 98%, approximately 97% to approximately 99%, approximately 97% to approximately 99.5%, approximately 97% to approximately 99.9%, approximately 98% to approximately 99%, approximately 98% to approximately 99.5%, approximately 98% to approximately 99.9%, approximately 99% to approximately 99.5%, approximately 99% to approximately 99.9%, or approximately 99.5% to approximately 99.9%.

[0164]

[0186] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence shown in any of SEQ ID NOs: 1 to 13. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence having 90 to 99% identity with the amino acid sequence shown in any of SEQ ID NOs: 1 to 13. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence with one, two, or up to three amino acid mutations compared to the amino acid sequence shown in any of SEQ ID NOs: 1 to 13.

[0165]

[0187] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence shown in any of SEQ ID NOs: 1-6. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence having 90-99% identity with the amino acid sequence shown in any of SEQ ID NOs: 1-6. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence with one, two, or up to three amino acid mutations compared to the amino acid sequence shown in any of SEQ ID NOs: 1-6.

[0166]

[0188] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises an amino acid sequence shown in any of SEQ ID NOs: 7-12. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises an amino acid sequence having 90-99% identity with the amino acid sequence shown in any of SEQ ID NOs: 7-12. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises an amino acid sequence with one, two, or up to three amino acid mutations compared to the amino acid sequence shown in any of SEQ ID NOs: 7-12.

[0167]

[0189] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises an amino acid sequence shown in any of SEQ ID NOs. 13-18. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises an amino acid sequence having 90-99% identity with the amino acid sequence shown in any of SEQ ID NOs. 13-18. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ and / or CD8+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises an amino acid sequence with one, two, or up to three amino acid mutations compared to the amino acid sequence shown in any of SEQ ID NOs. 13-18.

[0168]

[0190] In some embodiments, the overexpressed antigen may have lower expression levels in normal tissues and higher expression levels in tumors.

[0191] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, includes the amino acid sequence shown in SEQ ID NO: 1. In some embodiments, an epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, includes the amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 1. In some embodiments, an epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, includes the amino acid sequence shown in SEQ ID NO: 2. In some embodiments, the epitope, for example, a T cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a cancer-specific CD8+ epitope, includes an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 2. In some embodiments, the epitope, for example, a T cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a cancer-specific CD8+ epitope, includes an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 3. In some embodiments, the epitope, for example, a T cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a cancer-specific CD8+ epitope, includes an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 3. In some embodiments, the epitope, e.g., a T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises the amino acid sequence shown in SEQ ID NO: 4. In some embodiments, the epitope, e.g., a T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 4.In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, includes the amino acid sequence shown in SEQ ID NO: 5. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, includes the amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 5. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, includes the amino acid sequence shown in SEQ ID NO: 6. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 6.

[0169]

[0192] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, includes the amino acid sequence shown in SEQ ID NO: 7. In some embodiments, an epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, includes the amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 7. In some embodiments, an epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, includes the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the epitope, for example, a T cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a cancer-specific CD8+ epitope, includes an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 8. In some embodiments, the epitope, for example, a T cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a cancer-specific CD8+ epitope, includes an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 9. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises the amino acid sequence shown in SEQ ID NO: 10. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 10.In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises the amino acid sequence shown in SEQ ID NO: 11. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises the amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 11. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises the amino acid sequence shown in SEQ ID NO: 12. In some embodiments, the epitope, e.g., a T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 12.

[0170]

[0193] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, comprises the amino acid sequence shown in SEQ ID NO: 13. In some embodiments, an epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 13. In some embodiments, an epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises the amino acid sequence shown in SEQ ID NO: 14. In some embodiments, the epitope, for example, a T cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a cancer-specific CD8+ epitope, includes an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 14. In some embodiments, the epitope, for example, a T cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a cancer-specific CD8+ epitope, includes an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 15. In some embodiments, the epitope, for example, a T cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a cancer-specific CD8+ epitope, includes an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 15. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises the amino acid sequence shown in SEQ ID NO: 16. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 16.In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises the amino acid sequence shown in SEQ ID NO: 17. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises the amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 17. In some embodiments, the epitope, e.g., a T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises the amino acid sequence shown in SEQ ID NO: 18. In some embodiments, the epitope, e.g., a T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, comprises an amino acid sequence having 90-99% identity with the amino acid sequence of the epitope shown in SEQ ID NO: 18.

[0171]

[0194] In some embodiments, the recombinant polypeptide is 8 to 100 amino acids long. In some embodiments, the recombinant polypeptide is 5 to 15 amino acids long (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15). In some embodiments, the recombinant polypeptide is 8 to 12 amino acids long. In some embodiments, the recombinant polypeptide is 9 to 10 amino acids long. In some embodiments, the recombinant polypeptide is 8 amino acids long. In some embodiments, the recombinant polypeptide is 9 amino acids long. In some embodiments, the recombinant polypeptide is 10 amino acids long. In some embodiments, the recombinant polypeptide is 11 amino acids long. In some embodiments, the recombinant polypeptide is 12 amino acids long.

[0172]

[0195] In some embodiments, the recombinant polypeptide comprises two or more epitopes, for example, two, three, four, five, six, seven, eight, nine, ten, fifteen, twenty, or thirty epitopes, for example, T cell epitopes, for example, cancer-specific CD8+ and / or CD4+ epitopes, for example, cancer-specific CD8+ epitopes. In some embodiments, the recombinant polypeptide comprises at least two epitopes, for example, at least two T cell epitopes, for example, at least two cancer-specific CD8+ and / or CD4+ epitopes, for example, at least two cancer-specific CD8+ epitopes. In some embodiments, the epitopes are identical. In some embodiments, the epitopes are different. In non-limiting examples, in some embodiments, the recombinant polypeptide disclosed herein may comprise three epitopes, two of which are identical and one different, e.g., T cell epitopes, e.g., cancer-specific CD8+ and / or CD4+ epitopes, e.g., cancer-specific CD8+ epitope. In some embodiments, the first epitope, e.g., the first T cell epitope, e.g., the first cancer-specific CD8+ and / or CD4+ epitope, e.g., the first cancer-specific CD8+ epitope, comprises the amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, the second epitope, e.g., the second T cell epitope, e.g., the second cancer-specific CD8+ and / or CD4+ epitope, e.g., the second cancer-specific CD8+ epitope, comprises the amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, a third epitope, for example, a third T cell epitope, for example, a third cancer-specific CD8+ and / or CD4+ epitope, for example, a third cancer-specific CD8+ epitope, comprises an amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, a fourth epitope, for example, a fourth T cell epitope, for example, a fourth cancer-specific CD8+ and / or CD4+ epitope, for example, a fourth cancer-specific CD8+ epitope, comprises an amino acid sequence shown in any of SEQ ID NOs: 1 to 35.In some embodiments, a fifth epitope, for example, a fifth T cell epitope, for example, a fifth cancer-specific CD8+ and / or CD4+ epitope, for example, a fifth cancer-specific CD8+ epitope, includes an amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, a sixth epitope, for example, a sixth T cell epitope, for example, a sixth cancer-specific CD8+ and / or CD4+ epitope, for example, a sixth cancer-specific CD8+ epitope, includes an amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, a seventh epitope, for example, a seventh T cell epitope, for example, a seventh cancer-specific CD8+ and / or CD4+ epitope, for example, a seventh cancer-specific CD8+ epitope, includes an amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, the eighth epitope, for example, the eighth T cell epitope, for example, the eighth cancer-specific CD8+ and / or CD4+ epitope, for example, the eighth cancer-specific CD8+ epitope, includes the amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, the ninth epitope, for example, the ninth T cell epitope, for example, the ninth cancer-specific CD8+ and / or CD4+ epitope, for example, the ninth cancer-specific CD8+ epitope, includes the amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, the tenth epitope, for example, the tenth T cell epitope, for example, the tenth cancer-specific CD8+ and / or CD4+ epitope, for example, the tenth cancer-specific CD8+ epitope, includes the amino acid sequence shown in any of SEQ ID NOs: 1 to 35.

[0173]

[0196] In some embodiments, the epitope, e.g., T-cell epitope, e.g., cancer-specific CD8+ and / or CD4+ epitope, e.g., cancer-specific CD8+ epitope, is derived from tumor-associated antigens (TAAs). TAAs are antigenic products produced by cancer, and they provide biomarkers for tumor targeting and identification. TAAs can be broadly classified into abnormally expressed autoantigens, mutated autoantigens, and tumor-specific antigens. Therefore, TAAs may be upregulated or overexpressed in cancer cells. TAAs may be mutated within cancer cells. TAAs may be specific to cancer cells and may be expressed only within cancer cells, which are sometimes referred to as tumor-specific antigens. TAAs may be overexpressed antigens that have lower expression levels in normal tissues and higher expression levels in tumors.

[0174]

[0197] In some embodiments, TAAs are associated with aggressive tumor behavior and / or exhibit cancer-specific expression. In some embodiments, TAAs enhance the CD8+ T cell immune response. In some embodiments, tumor-associated antigens (TAAs) are tumor-specific antigens. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ epitope, is a cancer-testis antigen, e.g., NY-ESO-1 or a cancer-testis antigen that is a member of the MAGE-A family. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ epitope, is an endogenous retroviral protein.

[0175]

[0198] In some embodiments, TAAs are associated with aggressive tumor behavior and / or exhibit cancer-specific expression. In some embodiments, TAAs enhance the CD4+ T cell immune response. In some embodiments, tumor-associated antigens (TAAs) are tumor-specific antigens. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD4+ epitope, is a cancer-testis antigen, e.g., NY-ESO-1 or a cancer-testis antigen that is a member of the MAGE-A family. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD4+ epitope, is an endogenous retroviral protein.

[0176]

[0199] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is mutated in cancer cells. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is overexpressed in cancer cells. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is or includes a non-coding tumor-specific epitope. As used herein, the term “non-coding tumor-specific epitope” means a peptide found on cancer cells, where the peptide is derived from a nucleotide sequence that is epigenetically repressed in healthy cells. These peptide sequences are abnormally expressed in tumor cells.

[0177]

[0200] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is not a latent epitope. As used herein, “latent epitope” refers to an epitope that is not immunogenic in an immunocompetent individual. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is a latent epitope.

[0178]

[0201] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, may be a viral epitope optionally associated with virus-driven cancer. Virus-driven cancer may be a cancer caused by HPV (human papillomavirus), HTLV (human T-lymphotropic virus), or EBV (Epstein-Barr virus).

[0179]

[0202] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ epitope, is or is derived from a tumor-associated antigen selected from the group including TRP-1, CEA, TAG-72, 9D7, Ep-CAM, EphA3, telomerase, mesothelin, SAP-1, Melan-A / MART-1, tyrosinase, CLPP, cyclin-Al, cyclin-B1, MAGE-Al, MAGE-C1, MAGE-C2, SSX2, XAGE1b / GAGED2a, CD45, glypican-3, IGF2B3, kallikrein-4, KIF20A, lengsin, meloe, MUC5AC, survivin, PRAME, SSX-2, NY-ESO-1 / LAGE1, gp70, MC1R, TRP-1 / -2, O-catenin, BRCA1 / 2, or CDK4.

[0180]

[0203] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD4+ epitope, is or is derived from a tumor-associated antigen selected from the group including EBNA2, TARP, EBV LMP1, STEAP, WT1, HTLV-1 env, HER2 / neu, MAGE-A3, BRAF-V600E, NY-ESO-1, or gp100.

[0181]

[0204] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is an epitope associated with colorectal cancer, prostate cancer, esophageal cancer, liver cancer, kidney cancer, lung cancer, bladder cancer, breast cancer, pancreatic cancer, brain cancer, hepatocellular carcinoma, lymphoma, leukemia, gastric cancer, cervical cancer, ovarian cancer, thyroid cancer, melanoma, carcinoma, head and neck cancer, skin cancer, nasopharyngeal cancer, Epstein-Barr-driven cancer, human papillomavirus-driven cancer, or soft tissue sarcoma. In some embodiments, the cancer is a primary cancer. In some embodiments, the cancer is a recurrent and / or resistant cancer.

[0182]

[0205] In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is an esophageal cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a colorectal cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a prostate cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a liver cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a kidney cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a breast cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a bladder cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a pancreatic cancer-specific CD8+ T cell epitope.In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a lung cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a brain cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a hepatocellular carcinoma-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a gastric cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a cervical cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is an ovarian cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a thyroid cancer-specific CD8+ T cell epitope. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope is a melanoma-specific CD8+ T cell epitope.In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is a CD8+ T-cell epitope specific to lymphoma. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is a CD8+ T-cell epitope specific to leukemia. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is specific to carcinoma. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is a CD8+ T cell epitope specific to head and neck cancer. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is a CD8+ T cell epitope specific to skin cancer. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is a CD8+ T cell epitope specific to soft tissue sarcoma. In some embodiments, a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is a CD8+ T cell epitope specific to nasopharyngeal carcinoma.

[0183]

[0206] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, may be a private epitope. As used herein, the term “private epitope” means an epitope found exclusively in a single antigen in the cancer of a single person. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, may be a public epitope. As used herein, the term “public epitope” means an epitope found in the cancer of two or more people.

[0184]

[0207] In some embodiments, recombinant polypeptides comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, can induce an epitope-specific T cell response. In some embodiments, recombinant polypeptides comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, can induce an augmented memory CD8+ T cell response. In some embodiments, recombinant polypeptides comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, are not processed by proteasomes or immunoproteasomes in antigen-presenting cells (APCs).

[0185]

[0208] In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, may be a neoepitope. As used herein, the term “neoepitope” refers to an epitope resulting from a mutation in a tumor cell, in particular somatic mutations or passenger mutations, which can result in the production of a neoepitope. In some embodiments, a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, is not a neoepitope. In some embodiments, the recombinant polypeptide may be a glycoconjugate protein. In some embodiments, the recombinant polypeptide may be a ubiquitinated protein. In some embodiments, the recombinant polypeptide may be a phosphorylated protein. In some embodiments, the recombinant polypeptide may be an acetylated protein. In some embodiments, the recombinant polypeptide may be a methylated protein. In some embodiments, the recombinant polypeptide may include a signal peptide. As used herein, “signal peptide” refers to an amino acid sequence that can be ligated to the amino terminus of a recombinant polypeptide disclosed herein. Signal peptides typically direct the localization of proteins. Polynucleotides

[0209] In one embodiment, the Disclosure provides a polynucleotide encoding a recombinant polypeptide comprising a single epitope described herein, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope. In some embodiments, the polynucleotide encodes a recombinant polypeptide comprising a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, the polynucleotide encodes a recombinant polypeptide comprising a single epitope, e.g., a single T-cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising an amino acid sequence shown in any of SEQ ID NOs: 1 to 13. In some embodiments, the polynucleotide encodes a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising the amino acid sequence shown in any of SEQ ID NOs: 1 to 6. In some embodiments, the polynucleotide encodes a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising the amino acid sequence shown in any of SEQ ID NOs: 7 to 12. In some embodiments, the polynucleotide encodes a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, comprising the amino acid sequence shown in any of SEQ ID NOs: 13 to 18.

[0186]

[0210] Polynucleotides include, for example, genomic DNA, cDNA, RNA, such as mRNA, and DNA-RNA hybrid molecules. Polynucleotides may be naturally occurring, recombinant, or synthetic. In addition, polynucleotides may be single-stranded, double-stranded, or triple-stranded. In certain embodiments, polynucleotides may be modified. In the case of double-stranded polymers, “nucleic acid” may refer to either or both strands of the molecule. In some embodiments, polynucleotides are provided as DNA. In some embodiments, polynucleotides are provided as RNA, such as mRNA. In some embodiments, the polynucleotides disclosed herein include nucleic acid sequences shown in any of SEQ ID NOs. 52 to 86. In some embodiments, the polynucleotides disclosed herein include nucleic acid sequences shown in Table 2.

[0187] [Table 2]

[0188]

[0211] In some embodiments, the polynucleotides disclosed herein may be codon-optimized. Fractions in codon usage have been reported for numerous organisms, from viruses to eukaryotes. Since the genetic code is degenerate (i.e., each amino acid may be encoded by an average of three different codons), the DNA sequence may be modified by synonymous nucleotide substitutions without altering the amino acid sequence of the encoded protein. Such synonymous codon optimization is performed for the purpose of optimizing expression in a desired host, as described in scientific literature and patent documents. See U.S. Patents 5,786,464 and 6,114,14. In some embodiments, the nucleic acids described herein may be modified to improve cloning efficiency. In some embodiments, the nucleic acids described herein are subjected to codon optimization to increase the efficiency of gene expression; for example, SEQ ID NOs. 87-121 are subjected to codon optimization. In some embodiments, the polypeptides disclosed herein are encoded by polynucleotides whose sequences are codon-optimized for expression in mammalian cells, e.g., human cells. Exemplary codon-optimized sequences are disclosed in Table 3. In some embodiments, polypeptides encoded by codon-optimized nucleotides are expressed with increased expression in mammalian cells, such as human cells, compared to polypeptides encoded by non-codon-optimized polynucleotides.

[0189] [Table 3]

[0190]

[0212] In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 1 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 52 or 87. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 2 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 53 or 88. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 3 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 54 or 89. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 4 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 55 or 90. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 5 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 56 or 91. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 6 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 57 or 92. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 7 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 58 or 93. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 8 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 59 or 94. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 9 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 60 or 95. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 10 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 61 or 96. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 11 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 62 or 97. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 12 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 63 or 98.In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 13 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 64 or 99. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 14 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 65 or 100. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 15 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 66 or 101. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 16 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 67 or 102. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 17 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 68 or 103. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 18 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 69 or 104. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 19 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 70 or 105. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 20 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 71 or 106. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 21 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 72 or 107. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 22 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 73 or 108. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 23 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 74 or 109. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 24 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 75 or 110.In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 25 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 76 or 111. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 26 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 77 or 112. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 27 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 78 or 113. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 28 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 79 or 114. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 29 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 80 or 115. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 30 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 81 or 116. In some embodiments, a polypeptide containing the amino acid sequence shown in SEQ ID NO: 31 can be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 82 or 117. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 32 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 83 or 118. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 33 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 84 or 119. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 34 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 85 or 120. In some embodiments, a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 35 may be encoded by a polynucleotide having the sequence shown in SEQ ID NO: 86 or 121.

[0191]

[0213] In some embodiments, the polynucleotides disclosed herein can encode multiple epitopes, for example, two, three, four, five, six, seven, eight, nine, or ten epitopes. In some embodiments, the polynucleotide encodes multiple epitopes as distinct peptide epitopes. In some embodiments, the polynucleotide encodes multiple epitopes as a polyepitope peptide. In some embodiments, the epitopes are identical. In some embodiments, the epitopes are different. In some embodiments, the polynucleotide may include a sequence encoding at least two or more epitopes described herein, where both of its epitopes are the same epitope. In some embodiments, the polynucleotide may include a sequence encoding at least two or more epitopes described herein, where both of its epitopes are different epitopes. In non-limiting examples, in some embodiments, the polynucleotides disclosed herein can encode three epitopes, such as a T cell epitope, such as cancer-specific CD8+ and / or CD4+ epitopes, such as a cancer-specific CD8+ epitope, where two are the same epitope and one is a different epitope. In some embodiments, the polynucleotide comprises a nucleotide sequence encoding a spacer amino acid residue between the first and second epitopes.

[0192]

[0214] As will be recognized by those skilled in the art, in some embodiments, the nucleic acid includes a plasmid-derived regulatory sequence. The nucleic acid sequence may include, for example, one or more of a promoter sequence, a selection marker sequence, or a sequence that targets a locus. An example of a promoter capable of expressing a transgene in mammalian T cells is the EFla promoter (SEQ ID NO: 122). The native EFla promoter drives the expression of the alpha subunit of the elongation factor-1 complex, which results in the enzymatic delivery of aminoacyl-tRNA to the ribosome. In some embodiments, the polynucleotide includes the EFla promoter. Another example of a promoter is the pre-early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a strongly constitutive promoter sequence capable of driving high levels of expression of any polynucleotide sequence operably ligated to it. In some embodiments, the polynucleotide includes the CMV promoter. In some embodiments, the CMV promoter includes a sequence having 80-100% sequence identity with the sequence shown in SEQ ID NO: 36. In some embodiments, the polynucleotides of this disclosure may include, but are not limited to, promoters selected from the group consisting of the monkey virus 40 (SV40) initial promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) terminal repeat sequence (LTR) promoter, MoMuLV promoter, avian leukemia virus promoter, Epstein-Barr virus pre-initial promoter, Roussarcoma virus (RSV) promoter, actin promoter, myosin promoter, elongation factor-la promoter, hemoglobin promoter, and creatine kinase promoter. In some examples, the polynucleotide includes a sequence encoding a poly(A) tail. In some examples, the polynucleotide includes a 3'UTR sequence. In some examples, the polynucleotide includes a 5'UTR sequence.In some embodiments, the polynucleotides disclosed herein typically include a plurality of smaller, distinct nucleotide sequences that are heterogeneous and exhibit different measurable functions, a promoter sequence, a translation initiation sequence, a start codon, a polyadenylation sequence, a stop codon, an epitope, e.g., a T cell epitope, e.g., a cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope, e.g., a nucleotide sequence encoding an epitope including an amino acid sequence shown in any of SEQ ID NOs: 1 to 35, and / or a linker coding sequence. In some embodiments, the polynucleotides are codon-optimized, e.g., for optimal expression in mammalian cells, e.g., human cells.

[0193]

[0215] In some embodiments, a polynucleotide, such as RNA, such as mRNA, encoding an epitope, such as a T-cell epitope, such as a cancer-specific CD8+ and / or CD4+ epitope, such as a cancer-specific CD8+ epitope, is introduced into cells. In some embodiments, RNA transcribed in vitro may be introduced into cells as a form of transient transfection. In some embodiments, the polynucleotide may include some or all of the 5' and / or 3' untranslated regions (UTRs). The polynucleotide may include exons and introns. In some embodiments, the DNA used for PCR is a human nucleic acid sequence. In some embodiments, the DN used for PCR is a human nucleic acid sequence including the 5' and 3' UTRs. The DNA may, alternatively, be a naturally occurring artificial DNA sequence not normally expressed in organisms. An exemplary artificial DNA sequence is one which contains portions of genes that are ligated together to form an open reading frame encoding a fusion protein. The portions of DNA ligated together may be from a single organism or from two or more organisms.

[0194]

[0216] Any DNA polymerase useful for PCR may be used in the methods disclosed herein. Reagents and polymerases are commercially available from many sources. Chemical structures having the ability to enhance stability and / or translation efficiency may also be used. The RNA preferably has 5' and 3' UTRs. In some embodiments, the 5' UTR is 1 to 3,000 nucleotides long. The lengths of the 5' and 3' UTR sequences added to the coding region are not limited but may be modified by different methods, including designing primers for PCR that anneal to different regions of the UTR. Using this approach, those skilled in the art can modify the lengths of the 5' and 3' UTRs to achieve optimal translation efficiency after transfection of the transfected RNA.

[0195]

[0217] The 5' and 3' UTRs may be naturally occurring endogenous 5' and 3' UTRs for the polynucleotide of interest. Alternatively, non-endogenous UTR sequences for the polynucleotide of interest can be added by incorporating the UTR sequence into forward and reverse primers, or by any other modification of the template. The use of non-endogenous UTR sequences for the polynucleotide of interest may be useful for modifying RNA stability and / or translation efficiency. For example, it is known that AU-rich elements in 3' UTR sequences can reduce mRNA stability. Therefore, 3' UTRs may be selected or designed to increase the stability of transcribed RNA based on UTR properties known in the art.

[0196]

[0218] In some embodiments, the 5'UTR may contain a Kozak sequence of an endogenous nucleic acid. Alternatively, if a non-endogenous 5'UTR for the nucleic acid of interest is added by the PCR described above, the consensus Kozak sequence may be redesigned by adding a 5'UTR sequence. The Kozak sequence may increase the translation efficiency of some RNA transcripts. In some embodiments, the 5'UTR may be the 5'UTR of an RNA virus whose RNA genome is stable in cells. In some embodiments, various nucleotide analogs may be used in the 3' or 5'UTR to prevent exonuclease degradation of mRNA.

[0197]

[0219] In some embodiments, the polynucleotide further comprises a transcription promoter, such as the T7 polymerase promoter. Other useful promoters include, but are not limited to, the T3 and SP6 RNA polymerase promoters. Consensus nucleotide sequences for the T7, T3, and SP6 promoters are known in the art.

[0198]

[0220] In some embodiments, the polynucleotide, e.g., mRNA, has both a 5' cap and a 3' poly(A) tail, which determine ribosome binding, translation initiation, and mRNA stability in cells. In some embodiments, the polynucleotide includes a poly(A) tail, e.g., a poly(A) tail that provides stability to mRNA and / or reduces mRNA degradation. In some embodiments, the poly(A) tail is 100 to 5000 adenosines. In some embodiments, the polynucleotide may include an immunostimulatory sequence (e.g., ISS or CpG).

[0199]

[0221] In some embodiments, the polynucleotide may also contain an intra-sequence ribosome entry site (IRES) sequence. The IRES sequence may be a sequence from any viral chromosome or an artificially designed sequence that initiates cap-independent ribosome binding to mRNA and promotes the initiation of translation. It may also contain any solute suitable for cell electroporation, which may contain factors that promote cell permeability and viability, such as sugars, peptides, lipids, proteins, antioxidants, and surfactants.

[0200]

[0222] In some embodiments, the polynucleotides of this disclosure can encode any recombinant polypeptide described herein. For example, the polynucleotides can encode a recombinant polypeptide comprising a cancer-specific T cell epitope having at least about 90-100% sequence identity with SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, or SEQ ID NO: 35. In some embodiments, the polynucleotide can encode a recombinant polypeptide having at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, 12, 13, 14, or more amino acid residues from SEQ ID NOs: 1, SEQ ID NOs: 2, SEQ ID NOs: 3, SEQ ID NOs: 4, SEQ ID NOs: 5, SEQ ID NOs: 6, SEQ ID NOs: 7, SEQ ID NOs: 8, SEQ ID NOs: 9, SEQ ID NOs: 10, SEQ ID NOs: 11, SEQ ID NOs: 12, SEQ ID NOs: 13, SEQ ID NOs: 14, SEQ ID NOs: 15, SEQ ID NOs: 16, SEQ ID NOs: 17, SEQ ID NOs: 18, SEQ ID NOs: 19, SEQ ID NOs: 20, SEQ ID NOs: 21, SEQ ID NOs: 22, SEQ ID NOs: 23, SEQ ID NOs: 24, SEQ ID NOs: 25, SEQ ID NOs: 26, SEQ ID NOs: 27, SEQ ID NOs: 28, SEQ ID NOs: 29, SEQ ID NOs: 30, SEQ ID NOs: 31, SEQ ID NOs: 32, SEQ ID NOs: 33, SEQ ID NOs: 34, or SEQ ID NOs: 35.In some embodiments, the polynucleotide can encode a recombinant polypeptide having at least about one, two, three, four, five, six, seven, eight, nine, ten, eleven, 12, 13, 14, or more amino acid substitutions from SEQ ID NOs: 1, SEQ ID NOs: 2, SEQ ID NOs: 3, SEQ ID NOs: 4, SEQ ID NOs: 5, SEQ ID NOs: 6, SEQ ID NOs: 7, SEQ ID NOs: 8, SEQ ID NOs: 9, SEQ ID NOs: 10, SEQ ID NOs: 11, SEQ ID NOs: 12, SEQ ID NOs: 13, SEQ ID NOs: 14, SEQ ID NOs: 15, SEQ ID NOs: 16, SEQ ID NOs: 17, SEQ ID NOs: 18, SEQ ID NOs: 19, SEQ ID NOs: 20, SEQ ID NOs: 21, SEQ ID NOs: 22, SEQ ID NOs: 23, SEQ ID NOs: 24, SEQ ID NOs: 25, SEQ ID NOs: 26, SEQ ID NOs: 27, SEQ ID NOs: 28, SEQ ID NOs: 29, SEQ ID NOs: 30, SEQ ID NOs: 31, SEQ ID NOs: 32, SEQ ID NOs: 33, SEQ ID NOs: 34, or SEQ ID NOs: 35.

[0201]

[0223] In some embodiments, the polynucleotide encodes a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope that can bind to HLA molecules, e.g., HLA-A allele, e.g., HLA-B allele, e.g., HLA-C allele. In some embodiments, the HLA-A allele is HLA-A * 15 alleles (for example, HLA-A * 15:01 allele), HLA-A * 02 Alleles (e.g., HLA-A) * 02:01 Alleles, e.g., HLA-A * 02:05 Allele), HLA-A * 03 Alleles (e.g., HLA-A) * 03:01 Allele, HLA-A * 11 alleles (for example, HLA-A * 11:01 allele), or HLA-A * 31 alleles (for example, HLA-A * 31:01 allele). In some embodiments, the HLA-B allele is HLA-B * 07 Alleles (e.g., HLA-B) * 07:02 allele), HLA-B * 08 Alleles (e.g., HLA-B)* 08:01 allele), HLA-B * 15 alleles (for example, HLA-B) * 15:01 allele), HLA-B * 18 alleles (e.g., HLA-B) * 18:01 allele), HLA-B * 35 alleles (for example, HLA-B) * 35:01 allele, HLA-B * 35:03 allele), HLA-B * 40 alleles (for example, HLA-B) * 40:01 allele, HLA-B * 40:02 allele), or HLA-B * 44 alleles (for example, HLA-B) * 44:01 alleles, for example, HLA-B * 44:02 allele). In some embodiments, the HLA-C allele is HLA-C * 03 Alleles (e.g., HLA-C) * 03:04 alleles), or HLA-C * 07 Alleles (e.g., HLA-C) * 07:01 Alleles, e.g., HLA-C * (07:02 allele)

[0202]

[0224] The nucleotide sequences encoding an epitope, for example, a single T-cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a single cancer-specific CD8+ epitope, may consist of approximately 12 to approximately 45 base pairs, for example, approximately 15 to approximately 45 base pairs, approximately 18 to approximately 45 base pairs, approximately 21 to approximately 45 base pairs, or approximately 24 to approximately 45 base pairs.

[0203]

[0225] In some embodiments, the polynucleotides disclosed herein may be globular nucleic acids (SNAs). In some embodiments, the SNAs disclosed herein include nanoparticles conjugated with the polynucleotide sequences disclosed herein. In some embodiments, the SNAs disclosed herein include a monolayer liposome core. In some embodiments, the SNAs disclosed herein include one or more CpG oligonucleotides. In some embodiments, the SNAs disclosed herein include a polynucleotide, for example, a polynucleotide containing any of SEQ ID NOs. 52 to 121 encapsulated within a liposome core. In some embodiments, the polynucleotide, for example, a polynucleotide containing any of SEQ ID NOs. 52 to 121, is located on the surface of the SNAs disclosed herein. In some embodiments, the polynucleotide, for example, a polynucleotide containing any of SEQ ID NOs. 52 to 121, is chemically conjugated to the liposome surface of the SNAs disclosed herein. In some embodiments, the polynucleotide, for example, a polynucleotide containing any of SEQ ID NOs. 52 to 121, is hybridized to one or more CpG oligonucleotides and adsorbed onto the liposome surface. In some embodiments, the SNA disclosed herein may be a liposome SNA. vector

[0226] In one embodiment, the disclosure provides vectors, for example, vectors comprising recombinant polypeptides as described herein, and vectors comprising polynucleotides encoding recombinant polypeptides as described herein. In some embodiments, the vector is a viral vector. In some embodiments, the vector is a nonviral vector.

[0204]

[0227] In some embodiments, the viral vector may be an adenovirus vector, an adeno-associated virus (AAV) vector (e.g., AAV types 5 and 2), an alphavirus vector (e.g., Venezuelan encephalitis virus (VEE), Sindbis virus (SIN), Semryqui forest fever virus (SFV), and a VEE-SIN chimera), a flavivirus vector, a herpesvirus vector (e.g., a vector derived from a cytomegalovirus such as rhesus macaque cytomegalovirus (RhCMV)), an arenavirus vector (e.g., a lymphocytic choriomeningitis virus (LCMV) vector), a measles virus vector, a BCG vector, a vaccinia vector, a papillomavirus vector, and a poxvirus vector.

[0205]

[0228] In some embodiments, the vector includes an adenovirus vector. In some embodiments, the adenovirus vector may be or may be derived from a human adenovirus vector, a monkey adenovirus, or may include a type B adenovirus vector (e.g., Ad3, 11p, and 35), a type C adenovirus vector (e.g., Ad1, Ad2, Ad5, and Ad6), a type E adenovirus vector (e.g., Ad4), a type F adenovirus vector (e.g., Ad41), or a chimeric Ad5 vector (Ad5 / 11 and Ad5 / 35) containing Adll or Ad35 fibers, an adenovirus 6 vector, a PanAd3 vector, or an adenovirus C3 vector. In some embodiments, the adenovirus vector is human serotype 5 (AdHu5). In some embodiments, the vector may be an animal-derived adenovirus vector, e.g., derived from dogs, monkeys, and especially rhesus monkeys and chimpanzees. In some embodiments, the adenovirus vector may be a rare serotype vector derived from a non-human primate. Vectors derived from chimpanzees may be suitable for the vectors for this disclosure, and examples include, but are not limited to, ChAd63, ChAd3, and ChAdY25.

[0206]

[0229] In some embodiments, the vector is an adenovirus vector comprising a polynucleotide sequence encoding a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ epitope, and optionally, the vector is capable of inducing an augmented memory CD8+ T cell response. In some embodiments, the adenovirus vector comprises a polynucleotide sequence encoding a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ epitope, and the vector does not contain any additional cancer-specific CD8+ T cell epitopes. In some embodiments, the adenovirus vector comprises polynucleotide sequences encoding multiple epitopes, e.g., multiple T cell epitopes, e.g., multiple cancer-specific CD8+ epitopes.

[0207]

[0230] In some embodiments, the vector is or includes an AAV vector. In some embodiments, the AAV vector may be AAV1, AAV2, AAVS, or any combination thereof. In some embodiments, the AAV vector is selected for delivery to specific cells, tissues, or organs, for example, AAV serotypes 1, 2, 5, or hybrid capsids AAV1, AAV2, AAVS, or any combination thereof for targeting brain or nerve cells; AAV4 for targeting cardiac tissue; and AAV8 for delivery to the liver.

[0208]

[0231] In some embodiments, the vector may be modified to reduce its immunogenicity and improve its safety. In some embodiments, the vector may be replication-deficient. In some embodiments, the vector may lack E1 and E3 proteins.

[0209]

[0232] In some embodiments, the vector may be a recombinant (e.g., manipulated) adenovirus vector, the backbone of which is an adenovirus serotype 5 (Ad5) nucleic acid backbone. The recombinant adenovirus vector may contain a deletion or functional deletion of the E1 gene. The recombinant adenovirus vector may contain a deletion or functional deletion of the E3 gene. The recombinant adenovirus vector may contain deletions or functional deletions of both the E1 and E3 genes. The recombinant adenovirus vector may contain a transgene inserted into the locus of the deleted or functionally deleted E1 gene. The recombinant adenovirus vector may contain a transgene inserted into the locus of the deleted or functionally deleted E3 gene. The transgene insertion may contain a single cancer-specific CD8+ T cell epitope as described herein. The transgene insertion may contain a single cancer-specific CD4+ T cell epitope as described herein. The transgene insertion may contain two or more cancer-specific CD4+ T cell epitopes. The transgene insertion may contain cancer-specific CD8+ T cell epitopes and cancer-specific CD4+ T cell epitopes. A nucleic acid sequence encoding a single cancer-specific CD8+ T cell epitope may be inserted into the E1 region.

[0210]

[0233] In some embodiments, the vector further comprises a promoter. In some embodiments, the vector is a vector that is transcribed in vitro. In some embodiments, the vector may also contain a strong promoter, including, but are not limited to, the CMV promoter, RSV promoter, EFla promoter, monkey virus 40 (SV40) early promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) terminal repeat (LTR) promoter, MoMuLV promoter, avian leukemia virus promoter, Epstein-Barr virus pre-early promoter, Roussarcoma virus (RSV) promoter, and human gene promoters, including, but are not limited to, the actin promoter, myosin promoter, elongation factor-la promoter, hemoglobin promoter, and creatine kinase promoter. In some embodiments, the vector comprises the CMV promoter. An exemplary sequence for the CMV promoter is provided in SEQ ID NO: 31.

[0211]

[0234] In some embodiments, the vector may include an inductive promoter. Examples of inductive promoters include, but are not limited to, metallothionein promoters, glucocorticoid promoters, progesterone promoters, and tetracycline regulatory promoters.

[0212]

[0235] In some embodiments, the vectors disclosed herein may include additional promoter elements, such as enhancers and / or silencers.

[0213]

[0236] In some embodiments, the vector may include a TATA box. In some embodiments, the vector includes a translation initiation sequence, e.g., a Kossack sequence. The Kossack sequence has a consensus sequence (gcc)gccRccAUGG, and a preferred Kossack sequence is provided in SEQ ID NO: 37. In some embodiments, the vector includes a stop sequence and / or a polyadenylation sequence. A preferred polyadenylation sequence is provided in SEQ ID NO: 43. The AAV vector may include an inverted end repeat (ITR) sequence. A preferred 3'ITR sequence is provided in SEQ ID NO: 51. A preferred 5'ITR sequence is provided in SEQ ID NO: 50.

[0214]

[0237] In some embodiments, the viral vector includes a vector skeleton, a promoter region, and a nucleotide sequence encoding a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope.

[0215]

[0238] In some embodiments, the ITR sequence flanks to the encoded epitope, e.g., a T cell epitope, e.g., a cancer-specific CD8+ and / or CD4+ epitope, e.g., a cancer-specific CD8+ epitope. In some embodiments, there may be an ITR sequence located at 5' for the cancer-specific epitope. In some embodiments, there may be an ITR sequence located at 3' for the cancer-specific epitope. In some embodiments, there may be an ITR sequence located at 5' for the cancer-specific epitope and / or an ITR sequence located at 3' for the cancer-specific epitope. In some embodiments, the 5' ITR sequence may include SEQ ID NO: 50. In some embodiments, the 3' ITR sequence may include SEQ ID NO: 51. In some embodiments, the vector may include a 5' sequence for the cancer-specific epitope, e.g., SEQ ID NO: 44 or 47. In some embodiments, the vector may include a 3' sequence for the cancer-specific epitope, e.g., SEQ ID NO: 45 or 48. In some embodiments, a helper plasmid may be used to produce a viral vector containing a cancer-specific CD8+ T cell epitope. In some embodiments, one or more helper plasmids may be used to provide genes necessary for replication or packaging of the viral vector. In some embodiments, the helper plasmid encodes E2A, E4, and VA adenovirus proteins, as well as / or the rep and cap genes of AAV. In some embodiments, the vector may contain the sequence shown in SEQ ID NO: 46 or 49.

[0216]

[0239] In some embodiments, the vector may be modified to reduce its immunogenicity and improve its safety. In some embodiments, the vector may be replication-deficient. In some embodiments, the vector may lack E1 and E3 proteins. The vector may contain a 5' sequence for cancer-specific epitopes, e.g., SEQ ID NO: 44 or 47. The vector may contain a 3' sequence for cancer-specific epitopes, e.g., SEQ ID NO: 45 or 48.

[0217]

[0240] In some embodiments, the vector may also be derived from a retrovirus, such as a lentivirus. In some embodiments, the vectors disclosed herein, such as lentiviral vectors, can transduce non-proliferating cells, such as hepatocytes. In some embodiments, the vectors disclosed herein have low immunogenicity.

[0218]

[0241] The term "lentivirus" refers to a genus of retroviridae. Lentiviruses are unique among retroviruses in that they can infect non-dividing cells, and they can deliver a significant amount of genetic information to the host cell's DNA, making them one of the most efficient methods of gene delivery vectors. HIV, SIV, and FIV are all examples of lentiviruses.

[0219]

[0242] The term "lentiviral vector" refers specifically to vectors derived from at least a portion of a lentiviral genome, including the self-inactivating lentiviral vector provided in Milone et al., Mol.Ther.17(8):1453~1464 (2009). Other examples of lentiviral vectors that may be used in clinics include, but are not limited to, the LENTIVECTOR™ gene delivery technology from Oxford BioMedica and the LENTIMAX™ vector system from Lentigen. Non-clinical lentiviral vectors are also available and will be known to those skilled in the art.

[0220]

[0243] The recombinant polypeptides described herein may be used in polycistronic vectors or vectors that express several polypeptides in the same transcription unit. Such vectors may use intra-sequence ribosome entry sites (IRESs). Since IRESs do not function in all hosts and do not allow stoichiometric expression of multiple proteins, self-cleaving peptides may be used instead. For example, some viral peptides are cleaved during translation, allowing expression of multiple proteins from a single transcription unit. Such peptides include 2A peptides or 2A-like sequences derived from members of the Picornaviridae family. See, for example, Szymczak et al., 2004, Nature Biotechnology; 22:589-594. In some embodiments, the recombinant nucleic acids described herein encode a recombinant polypeptide in frame with a drug, and the two sequences are separated by a self-cleaving peptide, e.g., a 2A sequence or a T2A sequence.

[0221]

[0244] In some embodiments, the vectors disclosed herein may also contain a selection marker gene and / or a reporter gene, or both. In some embodiments, the selection marker may be delivered onto a separate piece of DNA and used in a co-transfection procedure. Both the selection marker and the reporter gene may be flanked with appropriate regulatory sequences to enable expression in host cells. Useful selection markers include, for example, antibiotic resistance genes, such as neo.

[0222]

[0245] Reporter genes are used to identify potentially transfected cells and to evaluate the functionality of regulatory sequences. Generally, a reporter gene is a gene that is not present in or expressed by the recipient organ or tissue, and whose expression is represented by some readily detectable characteristic, such as enzymatic activity, of a polypeptide. Reporter gene expression is assayed at a suitable time after the DNA has been introduced into recipient cells. Suitable reporter genes may include genes encoding luciferase, beta-galactosidase, chloramphenicol acetyltransferase, secreted alkaline phosphatase, or green fluorescent protein genes (e.g., Ui-Tei et al., 2000 FEBS Letters 479:79-82). Suitable expression systems are well known and can be prepared using known techniques or are commercially available. Generally, a construct having a minimal 5' flanking region exhibiting the highest level of reporter gene expression is identified as a promoter. Such a promoter region may be ligated to a reporter gene and used to assess a drug's ability to modulate promoter-driven transcription.

[0223]

[0246] In some embodiments, the vector is a non-viral vector, such as a plasmid, cosmid, virus-like particle, bacterial vector, artificial chromosome, liposome, lipid nanoparticle, or exosome. In some embodiments, the vector is a bacterial vector, such as attenuated Salmonella typhi, Salmonella tiphyta, Sigella, Bacillus, Lactobacillus, Calmette-Guérin bacillus (BCG), Escherichia coli, Vibrio cholerae, Campylobacter, or Listeria. In some embodiments, the vector is an attenuated and / or non-pathogenic vector.

[0224]

[0247] When nonviral vectors are used, an exemplary nonviral vector is a liposome. The use of lipid formulations is intended for the introduction of nucleic acids into host cells (in vitro, ex vivo, or in vivo). In another embodiment, nucleic acids may be associated with lipids. Lipid-associated nucleic acids may be encapsulated within the aqueous interior of a liposome, inserted into the lipid bilayer of a liposome, attached to a liposome via a linking molecule that associates with both the liposome and the oligonucleotide, captured within a liposome, complexed with a liposome, dispersed in a lipid-containing solution, mixed with lipids, combined with lipids, contained as a suspension in lipids, contained in or complexed with micelles, or otherwise associated with lipids. Compositions of associated lipids, lipid / DNA, or lipid / vectors in solution are not limited to any particular structure. For example, they may exist in a bilayer structure, as micelles, or in a "broken-down" structure. They may also simply be scattered in solution and may form aggregates that are not uniform in size or shape. Lipids are fatty substances that may be naturally occurring or synthetic. For example, lipids include naturally occurring lipid droplets in the cytoplasm, as well as a class of compounds containing long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, amino alcohols, and aldehydes.

[0225]

[0248] Lipids suitable for use can be obtained from commercial sources. For example, dimyristylphosphatidylcholine ("D1VIPC") can be obtained from Sigma, St. Louis, Mo.; dicetyl phosphate ("DCP") can be obtained from K & K Laboratories (Plainview, NY); cholesterol ("Choi") can be obtained from Calbiochem-Behring; and dimyristylphosphatidylglycerol ("DMPG") and other lipids may be obtained from Avanti Polar Lipids, Inc. (Birmingham, Ala.). Stock solutions of lipids in chloroform or chloroform / methanol can be stored at approximately -20°C. Chloroform is used as the sole solvent because it evaporates more readily than methanol. "Liposome" is a general term encompassing various monolayer and multilayer lipid vehicles formed by the creation of encapsulated lipid bilayers or aggregates. Liposomes can be characterized as having a vesicular structure with a phospholipid bilayer and an internal aqueous medium. Multilayer liposomes have multiple lipid layers separated by an aqueous medium. They spontaneously form when phospholipids are suspended in an excess aqueous solution. The lipid components undergo self-rearrangement before the formation of a closed structure and the capture of water and dissolved solute between the lipid bilayers (Ghosh et al., 1991 Glycobiology 5:505~10). However, compositions having structures in solution different from the usual vesicular structure are also included. For example, lipids may take the form of micellar structures or simply exist as heterogeneous aggregates of lipid molecules. Lipofectamine-nucleic acid complexes are also considered.

[0226]

[0249] In some embodiments, the polynucleotides of this disclosure can be cloned into many types of vectors. For example, the polynucleotides can be cloned into vectors, but are not limited to plasmids, phagemids, phage derivatives, viruses, and cosmids. In some embodiments, the polynucleotides can be cloned into vectors, but are not limited to expression vectors, replication vectors, probe-making vectors, and sequencing vectors. In some embodiments, the vectors disclosed herein include recombinant polypeptides disclosed herein, for example, a single epitope, for example, a single T-cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a single cancer-specific CD8+ epitope, for example, a single cancer-specific CD8+ epitope comprising the amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, the vectors disclosed herein include polynucleotides that encode a recombinant polypeptide, for example, a recombinant polypeptide, for example, a single epitope, for example, a single T cell epitope, for example, a single cancer-specific CD8+ and / or CD4+ epitope, for example, a recombinant polypeptide comprising a single cancer-specific CD8+ epitope comprising the amino acid sequence shown in any of SEQ ID NOs: 1 to 35.

[0227]

[0250] In some embodiments, the vector comprises a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD8+ epitope comprising the amino acid sequence shown in any of SEQ ID NOs: 1 to 35.

[0228]

[0251] In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 1. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 2. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 3. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 4. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 5. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 6. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 7. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 8. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 9. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 10. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 11. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 12. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 13. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 14. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 15. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 16.In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 17. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 18. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 19. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 20. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 21. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 22. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 23. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 24. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 25. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 26. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 27. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 28. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 29. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 30. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 31.In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 32. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 33. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 34. In some embodiments, the vector can encode a single cancer-specific CD8+ T cell epitope shown in SEQ ID NO: 35.

[0229]

[0252] In some embodiments, the vector encodes a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope comprising approximately 5 to approximately 15 amino acids, approximately 6 to approximately 15 amino acids, approximately 7 to approximately 15 amino acids, or approximately 8 to approximately 15 amino acids.

[0230]

[0253] In some embodiments, the vector encodes a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope selected from GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL. In some embodiments, the vector encodes two or more (e.g., two, three, four, five, six, or more) cancer-specific CD8+ and / or CD4+ T cell epitopes selected from GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL, or any combination thereof.

[0231]

[0254] In some embodiments, the vector encodes a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope selected from GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, LRPPTQEL, and RPASPRPAP. In some embodiments, the vector encodes two or more (e.g., two, three, four, five, six, or more) cancer-specific CD8+ and / or CD4+ T cell epitopes selected from GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, LRPPTQEL, and RPASPRPAP, or any combination thereof.

[0232]

[0255] In some embodiments, the vector is a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, LRPPTQE It encodes a single cancer-specific CD8+ epitope selected from L, RPASPRPAP, IATKIALQM, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, RVFTSSLKTK, RVLAKGLAK, GPRPSPTRSV, GQHLHLETF, SPRSPSPSL, VPQEAVRAPL, IYPFINSH, SPSSASLTL, LNKVKTSL, NLKTHLRL, FRGVFVHRY, TSGPVTEKY, and VVAAHLAGA. In some embodiments, the vector is GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, LRPPTQEL, RPASPRPAP, IATKIALQM, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, RVFTSSLKT The vector encodes two or more (e.g., two, three, four, five, six, or more) cancer-specific CD8+ and / or CD4+ T cell epitopes selected from K, RVLAKGLAK, GPRPSPTRSV, GQHLHLETF, SPRPSPSL, VPQEAVRAPL, IYPFINSH, SPSSASLTL, LNKVKTSL, NLKTHLRL, FRGVFVHRY, TSGPVTEKY, and VVAAHLAGA, or any combination thereof. In some embodiments, the vector encodes two or more (e.g., two, three, four, five, six, or more) cancer-specific CD8+ and / or CD4+ T cell epitopes selected from SEQ ID NOs: 1-6 or any combination thereof.In some embodiments, the vector encodes two or more (e.g., two, three, four, five, six, or more) cancer-specific CD8+ and / or CD4+ T cell epitopes selected from SEQ ID NOs. 7-12 or any combination thereof. In some embodiments, the vector encodes two or more (e.g., two, three, four, five, six, or more) cancer-specific CD8+ and / or CD4+ T cell epitopes selected from SEQ ID NOs. 13-18 or any combination thereof. In some embodiments, the vector comprises (i) a recombinant polypeptide containing a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide containing a cancer-specific CD8+ T cell epitope; where the cancer-specific CD8+ T cell epitope is GVYDGREHTV. In some embodiments, the vector comprises (i) a recombinant polypeptide consisting of a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide consisting of a cancer-specific CD8+ T cell epitope, where the cancer-specific CD8+ T cell epitope is GVYDGREHTV.

[0233]

[0256] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, where the cancer-specific CD8+ T cell epitope is KLVELEHTL.

[0234]

[0257] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, where the cancer-specific CD8+ T cell epitope is SPSSASLAL.

[0235]

[0258] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, where the cancer-specific CD8+ T cell epitope is KPRPDVTNEL.

[0236]

[0259] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, where the cancer-specific CD8+ T cell epitope is KEFAFLEHSL.

[0237]

[0260] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, where the cancer-specific CD8+ T cell epitope is HELGFKVVL.

[0238]

[0261] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KLVELEHTL.

[0239]

[0262] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and SPSSASLAL.

[0240]

[0263] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KPRPDVTNEL.

[0241]

[0264] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KEFAFLEHSL.

[0242]

[0265] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and HELGFKVVL.

[0243]

[0266] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and SPSSASLAL.

[0244]

[0267] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and KPRPDVTNEL.

[0245]

[0268] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and KEFAFLEHSL.

[0246]

[0269] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and HELGFKVVL.

[0247]

[0270] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and KPRPDVTNEL.

[0248]

[0271] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and KEFAFLEHSL.

[0249]

[0272] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and HELGFKVVL.

[0250]

[0273] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL and KEFAFLEHSL.

[0251]

[0274] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL and HELGFKVVL.

[0252]

[0275] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KEFAFLEHSL and HELGFKVVL.

[0253]

[0276] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and SPSSASLAL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and SPSSASLAL.

[0254]

[0277] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KEFAFLEHSL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KEFAFLEHSL.

[0255]

[0278] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KEFAFLEHSL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KEFAFLEHSL.

[0256]

[0279] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and HELGFKVVL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and HELGFKVVL.

[0257]

[0280] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KPRPDVTNEL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KPRPDVTNEL.

[0258]

[0281] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KEFAFLEHSL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KEFAFLEHSL.

[0259]

[0282] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and HELGFKVVL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and HELGFKVVL.

[0260]

[0283] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and KEFAFLEHSL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and KEFAFLEHSL.

[0261]

[0284] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and HELGFKVVL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and HELGFKVVL.

[0262]

[0285] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and HELGFKVVL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and HELGFKVVL.

[0263]

[0286] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KPRPDVTNEL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KPRPDVTNEL.

[0264]

[0287] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KEFAFLEHSL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KEFAFLEHSL.

[0265]

[0288] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and HELGFKVVL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and HELGFKVVL.

[0266]

[0289] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, KPRPDVTNEL, and KEFAFLEHSL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL, KPRPDVTNEL, and KEFAFLEHSL.

[0267]

[0290] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, KPRPDVTNEL, and HELGFKVVL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL, KPRPDVTNEL, and HELGFKVVL.

[0268]

[0291] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, KEFAFLEHSL, and HELGFKVVL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL, KPRPDVTNEL, and HELGFKVVL.

[0269]

[0292] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are SPSSASLAL, KPRPDVTNEL, and KEFAFLEHSL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL, KPRPDVTNEL, and KEFAFLEHSL.

[0270]

[0293] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are SPSSASLAL, KPRPDVTNEL, and HELGFKVVL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL, KPRPDVTNEL, and HELGFKVVL.

[0271]

[0294] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are SPSSASLAL, KEFAFLEHSL, and HELGFKVVL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL, KEFAFLEHSL, and HELGFKVVL.

[0272]

[0295] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least three cancer-specific CD8+ T cell epitopes are KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL. In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, wherein the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL.

[0273]

[0296] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, wherein the at least four cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, SPSSASLAL, and KPRPDVTNEL.

[0274]

[0297] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, wherein the at least four cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KLVELEHTL and SPSSASLAL and KEFAFLEHSL.

[0275]

[0298] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, wherein the at least four cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, SPSSASLAL, and HELGFKVVL.

[0276]

[0299] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, wherein the at least four cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, KPRPDVTNEL, and KEFAFLEHSL.

[0277]

[0300] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, wherein the at least four cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, KPRPDVTNEL, and HELGFKVVL.

[0278]

[0301] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, wherein the at least four cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL.

[0279]

[0302] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, wherein the at least four cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, KPRPDVTNEL, and KEFAFLEHSL.

[0280]

[0303] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, wherein the at least four cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, KPRPDVTNEL, and HELGFKVVL.

[0281]

[0304] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, wherein the at least four cancer-specific CD8+ T cell epitopes are KLVELEHTL, KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL.

[0282]

[0305] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least four cancer-specific CD8+ T cell epitopes, wherein the at least four cancer-specific CD8+ T cell epitopes are SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL.

[0283]

[0306] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, wherein the at least five cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, and KEFAFLEHSL.

[0284]

[0307] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, wherein the at least five cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, and HELGFKVVL.

[0285]

[0308] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, wherein the at least five cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL.

[0286]

[0309] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, wherein the at least five cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL.

[0287]

[0310] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, wherein the at least five cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL.

[0288]

[0311] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, wherein the at least five cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL.

[0289]

[0312] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least five cancer-specific CD8+ T cell epitopes, wherein the at least five cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, SPSSASLAL, KEFAFLEHSL, and HELGFKVVL.

[0290]

[0313] In some embodiments, the vector comprises (i) a recombinant polypeptide comprising at least six cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least six cancer-specific CD8+ T cell epitopes, wherein the at least six cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, and HELGFKVVL.

[0291]

[0314] In some embodiments, the vector includes a sequence having at least 90% sequence identity with sequence number 44 and a sequence having at least 90% sequence identity with sequence number 45. In some embodiments, the vector further includes a sequence having at least 90% sequence identity with sequence number 47 and a sequence having at least 90% sequence identity with sequence number 48.

[0292]

[0315] Cancer-specific CD8+ T cell epitopes may be determined using techniques known in the art, such as proteomics approaches, mass spectrometry approaches, genomic approaches, transcriptome analysis, bioinformatics approaches, and in silico methods. Those skilled in the art will be able to select suitable epitopes to be encoded in the vectors of this disclosure.

[0293]

[0316] Methods for introducing and expressing genes in cells are well known in the art. In the context of vectors, vectors can be readily introduced into host cells, such as mammalian cells, bacterial cells, yeast cells, or insect cells, by any method in the art. For example, vectors can be introduced into host cells by physical, chemical, or biochemical means.

[0294]

[0317] Physical methods for introducing polynucleotides into host cells include calcium phosphate precipitation, lipofection, particle guns, microinjection, electroporation, polymer encapsulation, peptide-mediated transfection, or biolistic particle delivery systems, such as "gene guns." Methods for producing cells containing vectors and / or exogenous nucleic acids are well known in the art. See, for example, Sambrook et al., 2012, Molecular Cloning: A Laboratory Manual, Vols. 1-4, Cold Spring Harbor Press, NY. A non-limiting example for introducing polynucleotides into host cells is calcium phosphate transfection.

[0295]

[0318] Biological methods for introducing target polynucleotides into host cells include the use of DNA and RNA vectors. Viral vectors, particularly retroviral vectors, have become the most widely used method for introducing genes into mammalian cells, such as human cells. Other viral vectors may be derived from lentiviruses, poxviruses, herpes simplex virus I, adenoviruses, and adeno-associated viruses (see, for example, U.S. Patents 5,350,674 and 5,585,362).

[0296]

[0319] Chemical methods for introducing polynucleotides into host cells include colloidal dispersions, such as macromolecular complexes, nanocapsules, nanoparticles, lipid nanoparticle conjugates, microspheres, beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An exemplary colloidal system for use as an in vitro and in vivo delivery vehicle is liposomes (e.g., artificial membrane vesicles). Other conventional methods of targeted delivery of nucleic acids are available, such as the delivery of polynucleotides using targeted nanoparticles or other suitable submicron-sized delivery systems.

[0297]

[0320] Various assays may be performed to confirm the presence of recombinant DNA sequences in host cells, regardless of the method used to introduce exogenous nucleic acids into host cells or otherwise expose cells to the inhibitors of this disclosure. Such assays include, for example, “molecular biological” assays well known to those skilled in the art, such as Southern blotting and Northern blotting, RT-PCR and PCR; “biochemical” assays, such as immunological means (ELISA and Western blotting); or assays described herein to identify agents within the scope of this disclosure, for example, to detect the presence or absence of specific peptides. Cells and populations of cells

[0321] In one embodiment, the Disclosure provides a cell comprising (i) a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope comprising the amino acid sequence shown in any of SEQ ID NOs: 1 to 35, or (ii) a cell comprising a polynucleotide encoding a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope comprising the amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, the cell may be an antigen-presenting cell (APC). In some embodiments, the cell may be a T cell stimulated with an APC as described herein. In some embodiments, the cell may comprise a CD8+ T cell. In some embodiments, the cell may comprise a CD4+ T cell. In some embodiments, the cell may be a growth-promoting memory T cell, e.g., a growth-promoting memory CD8+ T cell.

[0298]

[0322] In one embodiment, the disclosure provides a population of cells, wherein the cells in the population comprise (i) a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD8+ epitope comprising an amino acid sequence shown in any of SEQ ID NOs: 1 to 35, or (ii) a polynucleotide comprising a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD8+ epitope comprising an amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, the population of cells comprises a plurality of APCs. In some embodiments, the APC comprises (i) a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD8+ epitope comprising an amino acid sequence shown in any of SEQ ID NOs: 1 to 35, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ epitope, e.g., a single cancer-specific CD8+ epitope comprising an amino acid sequence shown in any of SEQ ID NOs: 1 to 35. In some embodiments, the cell population comprises a plurality of T cells stimulated with the APC described herein. In some embodiments, the cell population comprises a plurality of CD8+ T cells. In some embodiments, the cell population comprises a plurality of CD4+ T cells. In some embodiments, the cell population comprises a plurality of augmenting memory T cells, e.g., augmenting memory CD8+ T cells.

[0299]

[0323] The growth-promoting memory T cells described herein may be characterized by the presence of specific markers and cell surface markers. Methods for identifying and quantifying these markers are well known in the art. Examples of preferred methods, but are not limited to affinity-based separation methods, magnetic cell sorting techniques, and fluorescence-based cell sorting techniques, such as FACS (fluorescent cell sorting). Growth-promoting memory CD8+ T cells may also be characterized by the presence of several markers, examples, but are not limited to CX3CR1, KLRG-1, and CD44. Growth-promoting memory CD8+ T cells may also be characterized by the low expression of several markers, examples, but are not limited to CD62L, CD27, and CD127. The term "low expression" may refer to cells that do not express the marker, or it may refer to cells that have low expression of the marker compared to other cells in the sample. In some embodiments, the cell population comprises multiple augmentation memory T cells, for example, multiple augmentation memory CD8+ T cells, for example, multiple augmentation memory CD8+ T cells in the population that express one or more of CX3CR1, KLRG-1, and CD44, and / or have low expression of one or more of CD62L, CD27, and CD127.

[0300]

[0324] In some embodiments, the augmenting memory CD8+ T cells described herein are characterized by a marker selected from the group including CX3CR1+, KLRG-1+, CD44+, CD62L-, CCR7-, and CD45RA+ / -, where (+) indicates the presence of the marker, (-) indicates low or no expression of the marker, and (-) means that this low expression may be further indicated by "(low)". In some embodiments, the augmenting memory CD8+ T cells may be characterized by one or more markers selected from the group including CX3CR1+, KLRG-1+, CD44+, CD62L-, CCR7-, CD45RA+ / -, CD27-(low), and / or CD127-(low). In some embodiments, augmented memory CD8+ T cells may be characterized by the phenotype CX3CR1+, KLRG-1+, CD44+, CD62L-, CCR7-, CD45RA+ / -. Augmented memory CD8+ T cells may also be characterized by the phenotype CX3CR1+, KLRG-1+, CD44+, CD62L-, CCR7-, CD45RA+ / -, CD27-(low), CD127-(low). CD8+ T cells produced in the augmented memory response may have several other characteristics. For example, the cells may contain a transcriptional profile driven by Tbx21 (also referred to as T-bet). These cells show sustained expression of Tbx21. The cells may also show sustained expression of E2f2, a transcription factor generally involved in cell growth and proliferation. The cells may also lack or have low expression of the transcription factor Eomes. In some embodiments, cells comprising a recombinant polypeptide containing a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, include a transcriptional profile driven by Tbx21.In some embodiments, cells containing a recombinant polypeptide comprising a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, exhibit sustained expression of E2f2. In some embodiments, cells containing a single epitope, e.g., a single T cell epitope, e.g., a single cancer-specific CD8+ and / or CD4+ epitope, e.g., a single cancer-specific CD8+ epitope, exhibit low expression of the transcription factor Eomes.

[0301]

[0325] Proliferating memory CD8+ T cells may not exhibit classical contraction after antigen exposure. During classical memory evolution after antigen exposure, cells form a contracted central memory pool that constitutes <1% of total circulating CD8+ T cells. However, proliferating memory cells are maintained as a large pool of cells circulating in the blood. Therefore, the resulting proliferating memory CD8+ T cells account for approximately 0.01% to 20% of total CD8+ T cells. In some embodiments, the population of augmenting memory CD8+ T cells constitutes 0.01%, 0.02%, 0.03%, 0.05%, 0.07%, 0.09%, 0.1%, 0.3%, 0.5%, 0.7%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% of the total CD8+ T cells. In some embodiments, the population of augmented memory CD8+ T cells constitutes approximately 0.1% to 20%, 0.05% to 20%, 0.5% to 20%, 1% to 20%, 2% to 20%, 3% to 20%, 5% to 20%, 8% to 20%, 10% to 20%, 12% to 20%, 15% to 20%, or 18% to 20% of the total CD8+ T cells. In some embodiments, the population of augmented memory CD8+ T cells constitutes approximately 12% to 20% of the total CD8+ T cells.

[0302]

[0326] In some cases, a population of augmented memory CD8+ T cells may retain their effector memory phenotype. The resulting augmented memory CD8+ T cells may retain their memory effector phenotype for an extended period, where the effector phenotype is characterized by CD44+ and / or CD62L-. The augmented memory CD8+ T cells may retain their memory effector phenotype for up to 60 days after exposure to the vector of this disclosure. In some embodiments, a population of augmented memory CD8+ T cells may retain their memory effector phenotype for up to 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, or longer after exposure to the vector of this disclosure. In some embodiments, populations of augmenting memory CD8+ T cells may retain their memory effector phenotype for up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 17, 20, 25, 30, 35, 40, 50, 55, 60 days, or longer after exposure to the vector of the present disclosure.

[0303]

[0327] Proliferating memory CD8+ T cells may lack markers of exhaustion. T cell exhaustion can result from excessive TCR (T cell receptor) stimulation. Markers of T cell exhaustion may include, but are not limited to, upregulation of markers such as PD-1, Tim-3, or Lag-3. Therefore, in some embodiments, proliferating memory CD8+ T cells may lack, or demonstrate low expression of, markers selected from the group consisting of PD-1, Tim-3, and / or Lag-3, but are not limited to these.

[0304]

[0328] In some embodiments, the T cells described herein can demonstrate distinct transcriptional profiles from both a central memory T cell subset and an exhausted memory T cell subset. In some embodiments, the T cells described herein can demonstrate features such as enhanced redox resilience and resilience to oxidative stress, which may result from inherently low levels of reactive oxygen species. In some embodiments, the transcriptional profile is driven by the transcription factor Tbx21 with minimal contribution from Eomes. This results in a CD8+ T cell phenotype that is long-lived and present in numerous peripheral organs while retaining effector function. In some embodiments, antigen-specific augmentation memory CD8+ T cells are generated by one or more of the processing, presentation, and co-stimulatory conditions. In some embodiments, epitope processing occurs independently of presentation by proteasomes or immunoproteasomes and, optionally, by unusual non-hematopoietic APCs during later stages that may help preserve this phenotype. In some embodiments, the requirement for antigen processing can be bypassed by using (i) a recombinant polypeptide comprising one or more cancer-specific CD8+ epitopes, e.g., one or more T cell epitopes, e.g., one or more cancer-specific CD8+ epitopes, e.g., each comprising an amino acid sequence shown in any of SEQ ID NOs: 1 to 35, or (ii) a vector encoding a polynucleotide encoding a recombinant polypeptide comprising one or more cancer-specific CD8+ epitopes, e.g., one or more T cell epitopes, e.g., one or more cancer-specific CD8+ epitopes, e.g., each comprising an amino acid sequence shown in any of SEQ ID NOs: 1 to 35.In some embodiments, an augmented memory response is induced using (i) a recombinant polypeptide comprising one or more cancer-specific CD8+ epitopes, e.g., one or more T cell epitopes, e.g., one or more cancer-specific CD8+ epitopes, e.g., each comprising an amino acid sequence shown in any of SEQ ID NOs: 1 to 35, or (ii) a vector encoding a polynucleotide encoding a recombinant polypeptide comprising one or more cancer-specific CD8+ epitopes, e.g., one or more T cell epitopes, e.g., one or more cancer-specific CD8+ epitopes, e.g., each comprising an amino acid sequence shown in any of SEQ ID NOs: 1 to 35. composition

[0329] In one embodiment, the disclosure provides compositions, for example, therapeutic compositions. In some embodiments, the compositions, for example, therapeutic compositions induce an augmented memory CD8+ T cell response (i.e., a persistent, functional, and durable CD8+ T cell response). In some embodiments, the resulting pool of CD8+ T cells can resist exhaustion that may occur due to prolonged TCR stimulation.

[0305]

[0330] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is GVYDGREHTV. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is GVYDGREHTV. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is GVYDGREHTV, and the composition is HLA-A *02:01 For use in the treatment of cancer in subjects expressing MHC encoded by an allele. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is GVYDGREHTV, and the composition is HLA-A * 02:01 This is intended for use in the treatment of cancer in subjects expressing MHC encoded by alleles.

[0306]

[0331] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KLVELEHTL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KLVELEHTL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KLVELEHTL, and the composition is HLA-A *02:01 For use in the treatment of cancer in subjects expressing MHC encoded by an allele. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KLVELEHTL, and the composition is HLA-A * 02:01 This is intended for use in the treatment of cancer in subjects expressing MHC encoded by alleles.

[0307]

[0332] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is SPSSASLAL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is SPSSASLAL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is SPSSASLAL, and the composition comprises HLA-B * 07:02 allele and / or HLA-B *35:03 For use in the treatment of cancer in subjects expressing MHC encoded by an allele. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is SPSSASLAL, and the composition comprises HLA-B * 07:02 allele and / or HLA-B * 35:03 is intended for use in the treatment of cancer in subjects expressing MHC encoded by this allele.

[0308]

[0333] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KPRPDVTNEL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C *07:02 For use in the treatment of cancer in subjects expressing MHC encoded by an allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KPRPDVTNEL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof.

[0309]

[0334] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KEFAFLEHSL, and the composition comprises HLA-B * 40:01 allele, HLA-B * 40:02 allele, HLA-B *This is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 44:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KEFAFLEHSL, and the composition comprises HLA-B * This is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 44:02 allele. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KEFAFLEHSL, and the composition comprises HLA-B * 40:01 allele, HLA-B * 40:02 allele, HLA-B *This is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 44:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is KEFAFLEHSL, and the composition comprises HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 44:02 allele.

[0310]

[0335] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is HELGFKVVL, and the composition comprises HLA-B *18:01 For use in the treatment of cancer in subjects expressing MHC encoded by an allele. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is HELGFKVVL, and the composition comprises HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:01 allele. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is HELGFKVVL, and the composition comprises HLA-B * This is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 44:02 allele. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is HELGFKVVL, and the composition comprises HLA-B *18:01 For use in the treatment of cancer in subjects expressing MHC encoded by an allele. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is HELGFKVVL, and the composition comprises HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:01 allele. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is HELGFKVVL, and the composition comprises HLA-B * This is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 44:02 allele. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is HELGFKVVL, and the composition comprises HLA-B *It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele.

[0311]

[0336] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is NLRPPTQEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is NLRPPTQEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is NLRPPTQEL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 allele, HLA-C *03:03 For use in the treatment of cancer in subjects expressing MHC encoded by an allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, or (ii) a polynucleotide encoding a recombinant polypeptide comprising a cancer-specific CD8+ T cell epitope, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the cancer-specific CD8+ T cell epitope is NLRPPTQEL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 allele, HLA-C * 03:03 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof.

[0312]

[0337] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KLVELEHTL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KLVELEHTL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KLVELEHTL, and the composition is HLA-A *02:01 For use in the treatment of cancer in subjects expressing MHC encoded by alleles. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KLVELEHTL, and the composition comprises HLA-A * 02:01 This is intended for use in the treatment of cancer in subjects expressing MHC encoded by alleles.

[0313]

[0338] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and SPSSASLAL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and SPSSASLAL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and SPSSASLAL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B *35:03 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and SPSSASLAL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * This is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 35:03 allele, or any combination thereof.

[0314]

[0339] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KPRPDVTNEL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C *07:02 For use in the treatment of cancer in subjects expressing MHC encoded by an allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KPRPDVTNEL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof.

[0315]

[0340] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KEFAFLEHSL, and the composition is HLA-A * 02:01 Allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B *It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KEFAFLEHSL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0316]

[0341] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and HELGFKVVL, and the composition is HLA-A * 02:01 Allele, HLA-B * 18:01 Allele HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B *It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are GVYDGREHTV and HELGFKVVL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0317]

[0342] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and SPSSASLAL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and SPSSASLAL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and SPSSASLAL, and the composition is HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B *35:03 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and SPSSASLAL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * This is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 35:03 allele, or any combination thereof.

[0318]

[0343] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and KPRPDVTNEL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C *07:02 For use in the treatment of cancer in subjects expressing MHC encoded by an allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and KPRPDVTNEL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof.

[0319]

[0344] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and KEFAFLEHSL, and the composition is HLA-A * 02:01 Allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B *It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and KEFAFLEHSL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0320]

[0345] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and HELGFKVVL, and the composition is HLA-A * 02:01 Allele, HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B *It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KLVELEHTL and HELGFKVVL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0321]

[0346] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and KPRPDVTNEL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C *07:02 For use in the treatment of cancer in subjects expressing MHC encoded by an allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and KPRPDVTNEL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof.

[0322]

[0347] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and KEFAFLEHSL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B *It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and KEFAFLEHSL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0323]

[0348] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and HELGFKVVL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B *44:02 allele, HLA-B * This is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are SPSSASLAL and HELGFKVVL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0324]

[0349] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL and KEFAFLEHSL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * 35:03 allele, HLA-C *07:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL and KEFAFLEHSL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0325]

[0350] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL and HELGFKVVL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 allele, HLA-B * 18:01 allele, HLA-B *40:01 allele, HLA-B * 44:02 allele, HLA-B * This is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KPRPDVTNEL and HELGFKVVL, and the composition comprises HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 allele, HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0326]

[0351] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KEFAFLEHSL and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KEFAFLEHSL and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KEFAFLEHSL and HELGFKVVL, and the composition is HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B *It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least two cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least two cancer-specific CD8+ T cell epitopes are KEFAFLEHSL and HELGFKVVL, and the composition comprises HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0327]

[0352] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and SPSSASLAL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and SPSSASLAL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and SPSSASLAL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B *35:03 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by the APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and SPSSASLAL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * This is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 35:03 allele, or any combination thereof.

[0328]

[0353] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KPRPDVTNEL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C *07:02 For use in the treatment of cancer in subjects expressing MHC encoded by an allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KPRPDVTNEL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof.

[0329]

[0354] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KEFAFLEHSL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B *It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and KEFAFLEHSL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0330]

[0355] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and HELGFKVVL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B *44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KLVELEHTL, and HELGFKVVL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0331]

[0356] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KPRPDVTNEL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C *07:02 For use in the treatment of cancer in subjects expressing MHC encoded by an allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KPRPDVTNEL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof.

[0332]

[0357] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KEFAFLEHSL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B *40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and KEFAFLEHSL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0333]

[0358] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and HELGFKVVL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B* 18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, SPSSASLAL, and HELGFKVVL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B * 18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0334]

[0359] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and KEFAFLEHSL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C* 07:02 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and KEFAFLEHSL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0335]

[0360] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV, KPRPDVTNEL, and HELGFKVVL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C *07:02 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * 40:02 allele, HLA-B * 18:01 For use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are GVYDGREHTV and KPRPDVTNEL and HELGFKVVL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * 40:02 allele, HLA-B * 18:01 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof.

[0336]

[0361] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KPRPDVTNEL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KPRPDVTNEL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C *07:02 For use in the treatment of cancer in subjects expressing MHC encoded by an allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KPRPDVTNEL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-C * 07:02 is intended for use in the treatment of cancer in subjects expressing MHC encoded by the allele, or any combination thereof.

[0337]

[0362] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KEFAFLEHSL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KEFAFLEHSL, and the composition is HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B *40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and KEFAFLEHSL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof.

[0338]

[0363] In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and HELGFKVVL. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cells stimulated by an APC comprising (i), (ii), or (iii); where the at least three cancer-specific CD8+ T cell epitopes are KLVELEHTL, SPSSASLAL, and HELGFKVVL, and the composition comprises HLA-A * 02:01 Allele, HLA-B * 07:02 allele, HLA-B * 35:03 allele, HLA-B *18:01 allele, HLA-B * 40:01 allele, HLA-B * 44:02 allele, HLA-B * It is intended for use in the treatment of cancer in subjects expressing MHC encoded by the 40:02 allele, or any combination thereof. In some embodiments, the therapeutic composition comprises (i) a recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, or (ii) a polynucleotide encoding the recombinant polypeptide comprising at least three cancer-specific CD8+ T cell epitopes, (iii) a vector comprising (i) or (ii), (iv) an antigen-presenting cell (APC) comprising (i), (ii), or (iii), or (v) T cel...

Claims

1. A method for treating cancer in a subject requiring treatment for cancer, (i) Recombinant polypeptides containing cancer-specific CD8+ T cell epitopes; (ii) Polynucleotides encoding recombinant polypeptides containing cancer-specific CD8+ T cell epitopes; A vector containing (iii)(i) or (ii); Antigen-presenting cells (APCs) including (iv), (i), (ii), or (iii); or T cells stimulated by APCs containing (v)(i) or (ii) The step includes administering a composition containing the following: Here, (a) The subject is HLA-A ＊ 03:01 cells express MHC encoded by the allele, and the cancer-specific CD8+ T cell epitope is LKPDHIQR, KTEVHGRLK, RVSLPKLGYK, or SLFGARPGR; (b) The subject is HLA-B ＊ NLRPPTQEL is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 07:02 allele; (c) The subject is HLA-A ＊ The MHC expressed by the 31:01 allele, and the cancer-specific CD8+ T cell epitope is SLFGARPGR; and / or (d) The subject is HLA-B ＊ The MHC, encoded by the 07:02 allele, expresses LRPPTQEL, and the cancer-specific CD8+ T cell epitope is LRPPTQEL. method.

2. A method for treating cancer in a subject requiring treatment for cancer, (i) Recombinant polypeptides containing cancer-specific CD8+ T cell epitopes; (ii) Polynucleotides encoding recombinant polypeptides containing cancer-specific CD8+ T cell epitopes; A vector containing (iii)(i) or (ii); Antigen-presenting cells (APCs) including (iv), (i), (ii), or (iii); or T cells stimulated by APCs containing (v)(i) or (ii) The step includes administering a composition containing the following: Here, (a) The subject is HLA-A ＊ 02:01 The cells express MHC encoded by the allele, and the cancer-specific CD8+ T cell epitope is GVYDGREHTV or KLVELEHTL; (b) The subject is HLA-B ＊ The cells express MHC encoded by the 07:02 allele, and the cancer-specific CD8+ T cell epitope is either SPSSASLAL or KPRPDVTNEL; (c) The subject is HLA-B ＊ 35:3 expresses MHC encoded by the allele, and the cancer-specific CD8+ T cell epitope is SPSSASLAL or KPRPDVTNEL; (d) The subject is HLA-B ＊ The cells express MHC encoded by the 40:01 allele, and the cancer-specific CD8+ T cell epitope is either KEFAFLEHSL or HELGFKVVL; (e) the subject expresses an MHC encoded by an HLA-B ＊ 40:02 allele, and the cancer-specific CD8+ T cell epitope is KEFAFLEHSL or HELGFKVVL;​​ (f) The subject is HLA-B ＊ MHC expressed by 44:02 alleles, and the cancer-specific CD8+ T cell epitope is KEFAFLEHSL or HELGFKVVL; and / or (g) The subject is HLA-B ＊ The MHC expressed by the 18:01 allele, and the cancer-specific CD8+ T cell epitope is HELGFKVVL. method.

3. A method for treating cancer in a subject requiring treatment for cancer, (i) Recombinant polypeptides containing cancer-specific CD8+ T cell epitopes; (ii) Polynucleotides encoding recombinant polypeptides containing cancer-specific CD8+ T cell epitopes; A vector containing (iii)(i) or (ii); Antigen-presenting cells (APCs) including (iv), (i), (ii), or (iii); or T cells stimulated by APCs containing (v)(i) or (ii) The step includes administering a composition containing the following: Here, (a) The subject is HLA-A ＊ 03:01 cells express MHC encoded by this allele, and the cancer-specific CD8+ T cell epitope is SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, or RVFTSSLKTK; (b) The subject is HLA-B ＊ The MHC expressed by the 07:02 allele is the cancer-specific CD8+ T cell epitope RPASPRPAP; and / or (c) The subject is HLA-C ＊ The MHC expressed by the 03:04 allele, and the cancer-specific CD8+ T cell epitope is IATKIALQM. method.

4. A method for treating cancer in a subject requiring treatment for cancer, (i) Recombinant polypeptides containing cancer-specific CD8+ T cell epitopes, or (ii) Polynucleotides encoding recombinant polypeptides containing cancer-specific CD8+ T cell epitopes, A vector containing (iii)(i) or (ii); Antigen-presenting cells (APCs) including (iv), (i), (ii), or (iii); or T cells stimulated by APCs containing (v)(i) or (ii) The step includes administering a composition containing the following: Here, (a) The subject is HLA-A ＊ 02:01 The cells express MHC encoded by the allele, and the cancer-specific CD8+ T cell epitope is GVYDGREHTV or KLVELEHTL; (b) The subject is HLA-A ＊ 03:01 cells express MHC encoded by this allele, and the cancer-specific CD8+ T cell epitope is LKPDHIQR, RVSLPKLGYK, SLFGARPGR, or KTEVHGRLK; (c) The subject is HLA-A ＊ SLFGARPGR is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 31:01 allele; (d) The subject is HLA-B ＊ MHC is expressed by the 07:02 allele, and the cancer-specific CD8+ T cell epitope is SPSSASLAL, RPASPRPAP, NLRPPTQEL, or KPRPDVTNEL; (e) The subject is HLA-B ＊ 35:3 cells express MHC encoded by the allele and have cancer-specific CD8+ T cell epitopes SPSSASLAL, RPASPRPAP, NLRPPTQEL, or KPRPDVTNEL; (f) The subject is HLA-B ＊ The cells express MHC encoded by the 40:01 allele, and the cancer-specific CD8+ T cell epitope is either KEFAFLEHSL or HELGFKVVL; (g) The subject is HLA-B ＊ The cells express MHC encoded by the 40:02 allele, and the cancer-specific CD8+ T cell epitope is either KEFAFLEHSL or HELGFKVVL; (h) The subject is HLA-B ＊ The cells express MHC encoded by the 44:02 allele, and their cancer-specific CD8+ T cell epitope is either HELGFKVVL or KEFAFLEHSL; (i) The subject is HLA-B ＊ The MHC expressed by the 18:01 allele, and the cancer-specific CD8+ T cell epitope is HELGFKVVL; and / or (j) The target is HLA-C ＊ The cells express MHC encoded by the 07:02 allele, and the cancer-specific CD8+ T cell epitope is LRPPTQEL, NLRPPTQEL, or KPRPDVTNEL. method.

5. A method for treating cancer in a subject requiring treatment for cancer, (i) Recombinant polypeptides containing cancer-specific CD8+ T cell epitopes, or (ii) Polynucleotides encoding recombinant polypeptides containing cancer-specific CD8+ T cell epitopes, A vector containing (iii)(i) or (ii); Antigen-presenting cells (APCs) including (iv), (i), (ii), or (iii); or T cells stimulated by APCs containing (v)(i) or (ii) The step includes administering a composition containing the following: Here, (a) The subject is HLA-A ＊ GQHLHLETF is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 15:01 allele; (b) The subject is HLA-C ＊ IATKIALQM is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 03:04 allele; (c) The subject is HLA-C ＊ The cells express MHC encoded by the 07:02 allele, and the cancer-specific CD8+ T cell epitope is LRPPTQEL, NLRPPTQEL, or KPRPDVTNEL; (d) The subject is HLA-A ＊ 03:01 expresses MHC encoded by the allele, and the cancer-specific CD8+ T cell epitope is LKPDHIQR, RVSLPKLGYK, SLFGARPGR, KTEVHGRLK, RLHSFTLRQK, RVFTSSLKTK, RVLAKGLAK, KTPLHTLLK, KSYSKVLVR, MTYKIHFKK, or SLYQTIRLK; It is; (e) The subject is HLA-B ＊ Cells express MHC encoded by the 07:02 allele, and whose cancer-specific CD8+ T cell epitopes are SPSSASLAL, KPRPDVTNEL, NLRPPTQEL, RPASPRPAP, GPRPSPTRSV, SPRSPSPSL, VPQEAVRAPL, VPASPALSR, SPSAPPSL, SPTLNVSAL, or SPSSASLTL; (f) The subject is HLA-B ＊ The cells express MHC encoded by the 08:01 allele, and their cancer-specific CD8+ T cell epitope is LNKVKTSL or NLKTEILRL; (g) The subject is HLA-C ＊ The MHC expressed by the 07:01 allele is the cancer-specific CD8+ T cell epitope FRGVFVHRY; (h) The subject is HLA-B ＊ TSGPVTEKY is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 35:01 allele; (i) The subject is HLA-A ＊ VVAAHLAGA is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 02:05 allele; (j) The target is HLA-A ＊ 02:01 The cells express MHC encoded by the allele, and the cancer-specific CD8+ T cell epitope is GVYDGREHTV or KLVELEHTL; (k) The subject is HLA-B ＊ The cells express MHC encoded by the 40:01 allele, and the cancer-specific CD8+ T cell epitope is either KEFAFLEHSL or HELGFKVVL; (l) The subject is HLA-B ＊ The cells express MHC encoded by the 44:02 allele, and the cancer-specific CD8+ T cell epitope is either KEFAFLEHSL or HELGFKVVL; (m) The target is HLA-B ＊ HELGFKVVL is a cancer-specific CD8+ T cell epitope that expresses MHC encoded by the 18:01 allele; (n) The subject is HLA-A ＊ MHC, encoded by the 31:01 allele, expresses a cancer-specific CD8+ T cell epitope called SLFGARPGR. method.

6. The method according to any one of claims 1 to 5, wherein the cancer is esophageal cancer.

7. The method according to any one of claims 1 to 6, wherein the polynucleotide is a vector.

8. A composition comprising a vector encoding a recombinant polypeptide containing a cancer-specific CD8+ T cell epitope, including GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, LRPPTQEL, RPASPRPAP, IATKIALQM, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, RVFTSSLKTK, or any combination thereof.

9. The composition according to claim 8, wherein the recombinant polypeptide comprises a single cancer-specific CD8+ T cell epitope selected from the group consisting of GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, LRPPTQEL, or RPASPRPAP.

10. The composition according to claim 8 or 9, wherein the vector encodes a single cancer-specific CD8+ T cell epitope selected from the group consisting of GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, HELGFKVVL, LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, LRPPTQEL, or RPASPRPAP.

11. A composition comprising a vector encoding a recombinant polypeptide containing a cancer-specific CD8+ T cell epitope, including GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, or HELGFKVVL, or any combination thereof.

12. The composition according to any one of claims 8 to 11, wherein the recombinant polypeptide comprises a single cancer-specific CD8+ T cell epitope selected from the group consisting of GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, or HELGFKVVL.

13. The composition according to any one of claims 8 to 12, wherein the vector encodes a single cancer-specific CD8+ T cell epitope selected from the group consisting of GVYDGREHTV, KLVELEHTL, SPSSASLAL, KPRPDVTNEL, KEFAFLEHSL, or HELGFKVVL.

14. A composition comprising a vector encoding a recombinant polypeptide containing a cancer-specific CD8+ T cell epitope, including RPASPRPAP, IATKIALQM, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, or RVFTSSLKTK, or any combination thereof.

15. The composition according to any one of claims 8 to 10 or 14, wherein the recombinant polypeptide comprises a single cancer-specific CD8+ T cell epitope selected from the group consisting of RPASPRPAP, IATKIALQM, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, or RVFTSSLKTK.

16. The composition according to any one of claims 8 to 10 or 14 to 15, wherein the vector encodes a single cancer-specific CD8+ T cell epitope selected from the group consisting of RPASPRPAP, IATKIALQM, SLYQTIRLK, KTPLHTLLK, RLHSFTLRQK, or RVFTSSLKTK.

17. A composition comprising a vector encoding a recombinant polypeptide containing a cancer-specific CD8+ T cell epitope, including LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, or LRPPTQEL, or any combination thereof.

18. The composition according to any one of claims 8 to 10 or 17, wherein the recombinant polypeptide comprises a single cancer-specific CD8+ T cell epitope selected from the group consisting of LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, or LRPPTQEL.

19. The composition according to any one of claims 8 to 10 or 17 to 18, wherein the vector encodes a single cancer-specific CD8+ T cell epitope selected from the group consisting of LKPDHIQR, SLFGARPGR, KTEVHGRLK, NLRPPTQEL, RVSLPKLGYK, or LRPPTQEL.

20. A composition comprising a vector encoding a recombinant polypeptide containing a cancer-specific CD8+ T cell epitope, including RPASPRPAP, GQHLHLETF, IATKIALQM, SLYQTIRLK, GPRPSPTRSV, KTPLHTLLK, RLHSFTLRQK, VPQEAVRAPL, LNKVKTSL, NLKTHLRL, RVFTSSLKTK, RVLAKGLAK, FRGVFVHRY, SPSSASLTL, TSGPVTEKY, VVAAHLAGA, KSYSKVLVR, MTYKIFIFKK, VPASPALSR, SPSARPPSL, SPTLNVSAL, or any combination thereof.

21. The composition according to claim 20, wherein the recombinant polypeptide comprises a single cancer-specific CD8+ T cell epitope selected from the group consisting of RPASPRPAP, GQHLHLETF, IATKIALQM, SLYQTIRLK, GPRPSPTRSV, KTPLHTLLK, RLHSFTLRQK, VPQEAVRAPL, LNKVKTSL, NLKTHLRL, RVFTSSLKTK, RVLAKGLAK, FRGVFVHRY, SPSSASLTL, TSGPVTEKY, and VVAAHLAGA.

22. The composition according to claim 20 or 21, wherein the vector encodes a single cancer-specific CD8+ T cell epitope selected from the group consisting of RPASPRPAP, GQHLHLETF, IATKIALQM, SLYQTIRLK, GPRPSPTRSV, KTPLHTLLK, RLHSFTLRQK, VPQEAVRAPL, LNKVKTSL, NLKTHLRL, RVFTSSLKTK, RVLAKGLAK, FRGVFVHRY, SPSSASLTL, TSGPVTEKY, and VVAAHLAGA.

23. The method according to any one of claims 1 to 7, or the composition according to any one of claims 8 to 22, wherein the cancer-specific CD8+ T cell epitope is an esophageal cancer-specific CD8+ T cell epitope.

24. The method according to any one of claims 1 to 7, or the composition according to any one of claims 8 to 22, wherein the vector is a vector selected from plasmids, cosmids, bacterial vectors, viral vectors, artificial chromosomes, liposomes, lipid nanoparticles, or exosomes.

25. The method or composition according to claim 24, wherein the vector is a viral vector.

26. The method or composition according to claim 24 or 25, wherein the viral vector is an adenovirus vector.

27. The method or composition according to claim 26, wherein the adenovirus vector is an Ad5 vector.

28. The method or composition according to claim 24 or 25, wherein the viral vector is AAV.

29. The method according to any one of claims 1 to 7 and 23 to 28, or the composition according to any one of claims 8 to 28, wherein the recombinant polypeptide has a length of 8 to 100 amino acids.

30. The method according to any one of claims 1 to 7 and 23 to 29, wherein a cancer-specific CD8+ T cell epitope binds to an MHC class I molecule with a binding affinity of 500 nM or less, or the composition according to any one of claims 8 to 29.

31. The method according to any one of claims 1 to 7 and 23 to 29, wherein a cancer-specific CD8+ T cell epitope binds to an MHC class I molecule with a rank score of 2% or less, or the composition according to any one of claims 8 to 29.

32. The method according to any one of claims 1 to 7 and 23 to 31, or the composition according to any one of claims 8 to 31, wherein the recombinant polypeptide comprises a single cancer-specific CD8+ T cell epitope.

33. The method according to any one of claims 1 to 7 and 23 to 32, or the composition according to any one of claims 8 to 32, wherein the vector encodes a single cancer-specific CD8+ T cell epitope.

34. The method according to any one of claims 1 to 7 and 23 to 33, wherein the recombinant polypeptide comprises at least two different cancer-specific CD8+ T cell epitopes, or the composition according to any one of claims 8, 11, 14, 17, 20, and 23 to 33.

35. The method according to any one of claims 1 to 7 and 23 to 33, wherein the vector comprises at least two different recombinant polypeptides, each comprising a cancer-specific CD8+ T cell epitope, or the composition according to any one of claims 8, 11, 14, 17, 20, and 23 to 33.

36. The method according to any one of claims 1 to 7 and 23 to 35, or the composition according to any one of claims 8 to 35, wherein the composition further comprises an adjuvant.

37. The method according to any one of claims 1 to 7 and 23 to 36, further comprising the step of administering a second therapeutic agent.

38. The method according to claim 37, wherein the second therapeutic agent is selected from an immune checkpoint inhibitor, a chemotherapeutic agent, a small molecule inhibitor, or radiotherapy.

39. The method according to any one of claims 1 to 7 and 23 to 38, wherein the polynucleotide encoding the recombinant polypeptide is DNA.

40. The method according to any one of claims 1 to 7 and 23 to 38, wherein the polynucleotide encoding the recombinant polypeptide is RNA.

41. The method according to any one of claims 1 to 7, 23 to 28, and 39, wherein the polynucleotide encoding the recombinant polypeptide is messenger RNA (mRNA).

42. The method according to any one of claims 1 to 7 and 23 to 41, or the composition according to any one of claims 8 to 36, wherein the recombinant polypeptide has a length of 8 to 12 amino acids.

43. The method according to any one of claims 1 to 7 and 23 to 42, or the composition according to any one of claims 8 to 36 and 42, wherein the polynucleotide encoding the recombinant polypeptide is 9 to 10 amino acids long.

44. The method according to any one of claims 1 to 7 and 23 to 43, or the composition according to any one of claims 8 to 36 and 42 to 43, wherein the polynucleotide encoding the recombinant polypeptide is 9 amino acids long.

45. The method according to any one of claims 1 to 7 and 23 to 44, or the composition according to any one of claims 8 to 36 and 42 to 44, wherein the polynucleotide encoding the recombinant polypeptide is 10 amino acids long.

46. A composition according to any one of claims 8 to 36 and 42 to 45 for use in the treatment of cancer.

47. The composition according to claim 46, wherein the cancer is selected from colorectal cancer, prostate cancer, esophageal cancer, liver cancer, kidney cancer, lung cancer, bladder cancer, breast cancer, pancreatic cancer, brain cancer, hepatocellular carcinoma, lymphoma, leukemia, stomach cancer, cervical cancer, ovarian cancer, thyroid cancer, melanoma, carcinoma, head and neck cancer, skin cancer, nasopharyngeal cancer, Epstein-Barr-driven cancer, human papillomavirus-driven cancer, and soft tissue sarcoma.

48. The composition according to claim 45 or 46, wherein the cancer is esophageal cancer.

49. A method or composition according to any one of claims 1 to 48, wherein the composition induces an epitope-specific T cell response when administered to a subject.

50. The method or composition according to claim 49, wherein the response is an augmented memory CD8+ T cell response.

51. The method or composition according to any one of claims 1 to 50, wherein the composition is capable of inducing the production of CD8+ T cells characterized by markers comprising CX3CR1+, KLRG-1+, CD44+, and / or CD62L-.

52. The method or composition according to any one of claims 1 to 51, wherein the composition is capable of inducing the production of CD8+ T cells characterized by markers comprising CX3CR1+, KLRG-1+, CD44+, CD62L-, CD27-(low), and / or CD127-(low).

53. The method or composition according to any one of claims 1 to 52, wherein the composition is capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ / CD44+ T cells.

54. The method or composition according to any one of claims 1 to 53, wherein the composition is capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ / CD62L- T cells.

55. The method or composition according to any one of claims 1 to 54, wherein the composition is capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ / CD44+ / CD62L- T cells.

56. The method or composition according to any one of claims 1 to 55, wherein the composition is capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ / CD44+ / CD62L- / CD27- / CD127- T cells, CD8+ / CX3CR1+ / KLRG-1+ / CD44+ / CD62L- / CD27(low) / CD127- T cells, CD8+ / CX3CR1+ / KLRG-1+ / CD44+ / CD62L- / CD27- / CD127(low) T cells, or CD8+ / CX3CR1+ / KLRG-1+ / CD44+ / CD62L- / CD27(low) / CD127(low) T cells.

57. The method or composition according to any one of claims 1 to 56, wherein the composition is capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ T cells having sustained expression of Tbx21 and / or E2f2.

58. The method or composition according to any one of claims 1 to 57, wherein the composition is capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ T cells having low expression of Eomes.

59. The method or composition according to any one of claims 1 to 58, wherein the composition is capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ T cells, which constitute 0.01 percent to 20 percent of the total circulating CD8+ T cells in a subject.

60. The method or composition according to any one of claims 1 to 59, wherein the composition is capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ T cells that maintain a memory effector phenotype for at least 30 days.

61. The method or composition according to any one of claims 1 to 60, wherein the composition is capable of inducing the production of CD8+ / CX3CR1+ / KLRG-1+ T cells having low expression of PD-1, Tim-3, and / or Lag-3.

62. The method or composition according to any one of claims 1 to 61, wherein the composition is capable of inducing the production of an augmented memory CD8+ T cell response that can control tumor growth in a subject for longer than 50 days after administration of the composition.

63. The method or composition according to any one of claims 1 to 62, wherein the composition is capable of inducing the production of a nucleotide sequence encoding a cancer-specific CD8+ T cell epitope that encodes a polypeptide comprising a cancer-specific CD8+ T cell epitope, and the polypeptide comprising the cancer-specific CD8+ T cell epitope is not processed by the antigen-presenting cells of the subject when administered; and the pharmaceutical composition comprises a corresponding viral vector comprising a nucleotide sequence encoding a cancer-specific CD8+ T cell epitope that encodes a polypeptide comprising a cancer-specific CD8+ T cell epitope that is processed by the antigen-presenting cells of the subject when administered, wherein an augmented memory CD8+ T cell response is not induced in the subject.

64. The method or composition according to any one of claims 1 to 63, wherein the cancer-specific CD8+ T cell epitope is a viral antigen, a tumor-associated antigen overexpressed in cancer cells, or an antigen that is mutated in cancer cells.

65. The method or composition according to any one of claims 1 to 64, wherein the polypeptide comprising a cancer-specific CD8+ T cell epitope is not processed by the proteasome or immunoproteasome in antigen-presenting cells.

66. The method or composition according to any one of claims 1 to 64, wherein the viral vector comprises a CMV promoter and a TATA box.

67. The method or composition according to any one of claims 1 to 66, wherein the viral vector is a replication-deficient AdHu5 adenovirus vector.

68. The method or composition according to any one of claims 1 to 67, wherein the viral vector lacks sequences encoding El and E3 proteins.

69. The method or composition according to any one of claims 1 to 68, wherein the viral vector is AAV.

70. The method or composition according to any one of claims 1 to 69, wherein the composition comprises at least two viral vectors, each of which is a viral vector, and each of which encodes a different single cancer-specific CD8+ T cell epitope.

71. The method or composition according to any one of claims 1 to 70, wherein the composition comprises at least three viral vectors, each of which is a viral vector, and each of which encodes a different single cancer-specific CD8+ T cell epitope.

72. The method or composition according to any one of claims 1 to 71, wherein the composition comprises at least four viral vectors, each of which is a viral vector, and each of which encodes a different single cancer-specific CD8+ T cell epitope.

73. The method or composition according to any one of claims 1 to 72, wherein the composition comprises at least five viral vectors, each of which is a viral vector, and each of which encodes a different single cancer-specific CD8+ T cell epitope.

74. The method or composition according to any one of claims 1 to 73, wherein the composition comprises at least six viral vectors, each of which is a viral vector, and each of which encodes a different single cancer-specific CD8+ T cell epitope.

75. A pharmaceutical composition comprising the composition according to any one of claims 1 to 74, and a pharmaceutically acceptable carrier, diluent, excipient, or adjuvant.

76. A method for treating or preventing cancer, comprising the step of administering a therapeutically effective amount of the pharmaceutical composition according to claim 75 to a subject in need of treating or preventing cancer.

77. A method for inducing an augmented memory CD8+ T cell response, comprising the step of administering a therapeutically effective amount of the pharmaceutical composition according to claim 75 to a subject requiring induction of an augmented memory CD8+ T cell response, wherein the augmented memory CD8+ T cell response comprises the production of CD8+ / CX3CR1+ / KLRG-1+ T cells.

78. The method or composition according to any one of claims 1 to 77, wherein the vector or composition is administered intravenously or intramuscularly.

79. The method or composition according to any one of claims 1 to 78, wherein the composition is administered as a single dose.

80. The method or composition according to any one of claims 1 to 79, wherein the composition is administered in multiple doses.

81. The method or composition according to any one of claims 1 to 80, wherein the composition is administered in two doses.

82. The method or composition according to claim 80 or 81, wherein the first dose is the prime dose.

83. The method or composition according to any one of claims 80 to 82, wherein a first dose induces a measurable immune response in a subject compared to an immune response in the subject in the absence of administration of the first dose.

84. The method or composition according to any one of claims 80 to 83, wherein the second dose is a booster dose.

85. The method or composition according to any one of claims 80 to 84, wherein a second dose induces a measurable immune response in a subject compared to an immune response in the subject in the absence of administration of the second dose.

86. The method or composition according to any one of claims 80 to 85, wherein the second dose is administered 24 weeks after the first dose.

87. The method or composition according to any one of claims 80 to 86, wherein the second dose induces an immune response at least twice as high as the immune response induced by the first dose.

88. The method or composition according to any one of claims 1 to 87, wherein the composition is administered prophylactically to a subject.

89. The method or composition according to any one of claims 1 to 88, wherein the composition is administered in combination with a second therapeutic agent.

90. The method or composition according to any one of claims 1 to 89, wherein the second therapeutic agent is selected from an immune checkpoint inhibitor, a chemotherapeutic agent, a small molecule inhibitor, or radiotherapy.

91. The method or composition according to claim 90, wherein the immune checkpoint inhibitor is an inhibitor of an immune checkpoint protein selected from the group consisting of CTLA-4, PD-1, PD-L1, PD-L2, TIM3, LAG-3, B7-H3, B7-H4, B7-H6, A2aR, BTLA, GALS, and IDO.

92. A method for producing the composition according to any one of claims 1 to 91, (i) A step of synthesizing a nucleotide sequence encoding a single cancer-specific CD8+ T cell epitope as sense and antisense primers, (ii) The step of cloning the nucleotide sequence encoding a single cancer-specific CD8+ T cell epitope synthesized in (i) into the first plasmid, (iii) A step of cloning a sequence containing a nucleotide sequence encoding a single cancer-specific CD8+ T cell epitope derived from the first plasmid of (iii) into a second vector containing adenovirus DNA. A method that includes this.

93. A method for treating or preventing cancer, comprising the step of administering a composition according to any one of claims 1 to 92, comprising at least two viral vectors present in the pharmaceutical composition, to a subject in need of treatment or prevention of cancer, thereby treating cancer in the subject.

94. The method or composition according to any one of claims 1 to 93, wherein the viral vector comprises a sequence having at least 90% sequence identity with SEQ ID NO: 44 and a sequence having at least 90% sequence identity with SEQ ID NO:

45.

95. The method or composition according to any one of the preceding claims, wherein the viral vector further comprises a sequence having at least 90% sequence identity with SEQ ID NO: 47 and a sequence having at least 90% sequence identity with SEQ ID NO:

48.

96. A method for treating cancer in a subject requiring treatment for cancer, (i) Recombinant polypeptides comprising cancer-specific CD8+ T cell epitopes, wherein the recombinant polypeptide binds to the TCR; (ii) A polynucleotide encoding a recombinant polypeptide containing a cancer-specific CD8+ T cell epitope, wherein the recombinant polypeptide encoded by the polynucleotide binds to a TCR; Cells containing recombinant polypeptides or polynucleotides of (iii)(i) or (ii); (iv) Recombinant polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain binds to a cancer-specific CD8+ T cell epitope; (v) A polynucleotide encoding a recombinant polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain binds to a cancer-specific CD8+ T cell epitope; or Cells containing recombinant polypeptides or polynucleotides of (vi), (iii), or (iv). A method comprising the step of administering a composition containing the above to a target.

97. The viral vector contains one or more of the following: a promoter, a translation start sequence, a start codon, a T cell epitope codon, a stop codon, and a polyadenylation sequence. (i) The promoter is selected from the CMV promoter, the RSV promoter, and the EFla promoter, and the promoter may include the sequence shown in SEQ ID NO: 36; (ii) The translation initiation sequence includes the sequence shown in SEQ ID NO: 37; and the polyadenylation sequence includes the sequence shown in SEQ ID NO:

43. A viral vector according to any one of claims 1 to 96.

98. A viral vector according to any one of claims 1 to 97, wherein the vector comprises a deletion or functional deletion in the E1 gene at the locus of the E1 gene.

99. The viral vector according to claim 98, wherein the vector includes an introduced gene insertion at the locus of the E1 gene.

100. The method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence GVYDGREHTV, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KLVELEHTL; the subject is HLA-A ＊ The method according to claim 2, wherein MHC encoded by the 02:01 allele is expressed.

101. The method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence SPSSASLAL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KPRPDVTNEL; the subject is HLA-B ＊ The method according to claim 2, wherein MHC encoded by the 07:02 allele is expressed.

102. The method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence SPSSASLAL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KPRPDVTNEL; the subject is HLA-B ＊ The method according to claim 2, wherein MHC encoded by the 35:03 allele is expressed.

103. The method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KEFAFLEHSL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence HELGFKVVL; the subject is HLA-B ＊ The method according to claim 2, wherein MHC encoded by the 40:01 allele is expressed.

104. The method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KEFAFLEHSL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence HELGFKVVL; the subject is HLA-B ＊ The method according to claim 2, wherein MHC encoded by the 40:02 allele is expressed.

105. The method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KEFAFLEHSL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence HELGFKVVL; the subject is HLA-B ＊ The method according to claim 2, wherein MHC encoded by the 44:02 allele is expressed.

106. The method comprises the step of administering to a subject (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KEFAFLEHSL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence HELGFKVVL; the subject is HLA-B ＊ The method according to claim 2, wherein MHC encoded by the 18:01 allele is expressed.

107. The composition according to claim 8, wherein the composition comprises (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having sequence GVYDGREHTV, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having sequence KLVELEHTL.

108. The composition according to claim 107 for use in the treatment of esophageal cancer in a subject.

109. The target is HLA-A ＊ The composition according to claim 107, for use in the treatment of esophageal cancer in a subject, expressing MHC encoded by the 02:01 allele.

110. The composition according to claim 8, wherein the composition comprises (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence SPSSASLAL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having the sequence KPRPDVTNEL.

111. The composition according to claim 110 for use in the treatment of esophageal cancer in a subject.

112. The target is HLA-B ＊ The composition according to claim 110, for use in the treatment of esophageal cancer in a subject, expressing MHC encoded by the 07:02 allele.

113. The target is HLA-B ＊ The composition according to claim 110, for use in the treatment of esophageal cancer in a subject, expressing MHC encoded by the 35:03 allele.

114. The composition according to claim 8, wherein the composition comprises (i) a first adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having sequence KEFAFLEHSL, and (ii) a second adenovirus vector comprising a sequence encoding a single cancer-specific CD8+ T cell epitope having sequence HELGFKVVL.

115. The composition according to claim 114 for use in the treatment of esophageal cancer in a subject.

116. The target is HLA-B ＊ The composition according to claim 114, for use in the treatment of esophageal cancer in a subject, expressing MHC encoded by a 40:01 allele.

117. The target is HLA-B ＊ The composition according to claim 114, for use in the treatment of esophageal cancer in a subject, expressing MHC encoded by 40:02 alleles.

118. The target is HLA-B ＊ The composition according to claim 114, for use in the treatment of esophageal cancer in a subject, expressing MHC encoded by 44:02 alleles.

119. The target is HLA-B ＊ The composition according to claim 114, for use in the treatment of esophageal cancer in a subject, expressing MHC encoded by the 18:01 allele.

120. A method according to any one of claims 100 to 106, or a composition according to any one of claims 107 to 119, wherein the first adenovirus vector is an Ad5 adenovirus vector and / or the second adenovirus vector is an Ad5 adenovirus vector.

121. A method according to any one of claims 100 to 106 and 120, wherein the first adenovirus vector comprises a deletion or functional deletion in the E1 gene at the E1 gene locus, and / or the second adenovirus vector comprises a deletion or functional deletion in the E1 gene at the E1 gene locus, or the composition according to any one of claims 107 to 120.

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