METHOD FOR T CELL ACTIVATION FOR CANCER TREATMENT

Abstract

The antigen-presenting cell loaded with the cancer-specific tumor antigen epitope provided in the present invention, that is, a dendritic cell enables rapid and effective induction of differentiation and proliferation of cancer antigen-specific T cells, preferably memory T cells, and the memory T cells thus activated can treat a cancerous or neoplastic condition or prevent recurrence, progression, or metastasis of cancer while avoiding the defense mechanism of cancer cells.

Claims

1-36. (canceled)

37. An antigen-presenting cell loaded with an Epstein-Barr virus (EBV)-positive cancer-specific tumor antigen epitope, wherein the epitope is derived from Epstein-Barr virus latent membrane protein 2 (LMP2a) or Epstein-Barr nuclear antigen 1 (EBNA-1).

38. The antigen-presenting cell of claim 37, wherein the epitope is derived from LMP2a and comprises an amino acid sequence as set forth in any one of SEQ ID NOs: 1 to 3, SEQ ID NOs: 7 to 9, SEQ ID NOs: 11 to 15, SEQ ID NOs: 19 to 21, SEQ ID NO: 23 to 39, SEQ ID NO: 43 to 45, SEQ ID NO: 47 to 51, SEQ ID NO: 55 to 57, SEQ ID NO: 59 to 63, SEQ ID NO: 67 to 69, SEQ ID NO: 71 to 75, SEQ ID NO: 79 to 81, SEQ ID NO: 83 to 111, SEQ ID NOs: 115 to 117, and SEQ ID NOs: 119 to 122.

39. The antigen-presenting cell of claim 37, wherein the epitope is derived from EBNA-1 and comprises an amino acid sequence as set forth in any one of SEQ ID NOs: 124 to 127, SEQ ID NOs: 129 to 149, SEQ ID NOs: 151 to 154, SEQ ID NOs: 156 to 158, SEQ ID NOs: 160 to 163, SEQ ID NOs: 165 to 167, SEQ ID NOs: 168 to 176, SEQ ID NOs: 178 to 181, SEQ ID NOs: 183 to 194, and SEQ ID NOs: 196 to 199.

40. The antigen-presenting cell of claim 37, wherein the Epstein-Barr virus (EBV)-positive cancer is selected from the group consisting of EBV-positive gastric cancer, EBV-positive cervical cancer, EBV-positive Burkitt's lymphoma, EBV-positive T cell lymphoma, EBV-positive breast cancer, EBV-positive leiomyosarcoma, EBV-positive smooth muscle tumor, EBV-positive Hodgkin lymphoma, EBV-positive nasopharyngeal cancer, and EBV-positive post-transplant lymphoproliferative disorder (PTLD).

41. The antigen-presenting cell of claim 37, wherein the antigen-presenting cell promotes proliferation or differentiation of T cells and is selected from the group consisting of dendritic cell, B cell, and macrophage.

42. A method for producing the antigen-presenting cell of claim 37, comprising pulsing an antigen-presenting cell with the epitope.

43. The method of claim 42, wherein the antigen-presenting cell is mixed with a solution containing the epitope.

44. A method for producing the antigen-presenting cell of claim 37, comprising transfecting an antigen-presenting cell with an expression vector via nucleofection, wherein the expression vector comprises a nucleic acid sequence encoding the epitope.

45. A method for producing the antigen-presenting cell of claim 37, comprising loading an antigen-presenting cell with the epitope using a fusion protein that comprises the epitope.

46. The method of claim 45, wherein the fusion protein further comprises a dendritic cell-specific antibody or a fragment thereof.

47. The method of claim 46, wherein the epitope is conjugated to the dendritic cell-specific antibody or a fragment thereof.

48. The method of claim 46, wherein the dendritic cell-specific antibody is an antibody specific for DCIR, MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11b, CD14, CD15, CD16, CD19, CD20, CD29, CD31, CD40, CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, DC-ASPGR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, Clec9A, 33D1, mannose receptor, Langerin, DECTIN-1, B7-1, B7-2, IFN-γ receptor, IL-2 receptor, ICAM-1, Fcγ receptor, LOX-1, or ASPGR.

49. A method for preventing or treating an Epstein-Barr virus (EBV)-positive cancer, the method comprising administering a therapeutically effective amount of the antigen-presenting cell of claim 37 to a target individual.

50. A method for activating T cells for cancer treatment or prevention, the method comprising co-culturing T cells with the antigen-presenting cell of claim 37.

51. The method of claim 50, wherein the T cells are obtained from peripheral blood mononuclear cells (PBMCs) of a target individual and include one or more selected from the group consisting of cytotoxic T cells, helper T cells, natural killer T cells, γδ T cells, regulatory T cells, and memory T cells.

52. The method of claim 50, wherein the co-culturing is performed with addition of interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-7 (IL-7), interleukin-15 (IL-15), interleukin-21 (IL-21), or a combination thereof.

53. The method of claim 50, wherein the co-culturing is performed with addition of a fusion protein that contains a cytokine and an immunoglobulin heavy chain constant region.

54. The method of claim 53, wherein the cytokine is interferon-γ (IFN-γ), interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-12 (IL-12), IL-18, tumor necrosis factor (TNF), or granulocyte macrophage colony stimulating factor (GMCSF).

55. The method of claim 50, wherein the co-culturing is performed with addition of a fusion protein that contains ligand of CD27, CXCR3, or CD62L, and an immunoglobulin heavy chain constant region.

56. A method for preventing or treating an Epstein-Barr virus (EBV)-positive cancer, the method comprising administering a therapeutically effective amount of T cells to a target individual, wherein the T cells are activated by the method of claim 50.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0094] FIGS. 1 to 4 illustrate results obtained by performing ELISPOT on each of LMP2a-derived epitopes in order to check the degree of T cell activation caused by the epitope in normal human-derived PBMCs, according to an embodiment of the present invention.

[0095] FIGS. 5 to 8 illustrate results obtained by performing ELISPOT on each of EBNA-1-derived epitopes in order to check the degree of T cell activation caused by the epitope in normal human-derived PBMCs, according to an embodiment of the present invention.

DETAILED DESCRIPTION OF INVENTION

[0096] According to an embodiment of the present invention, there is provided an Epstein-Barr virus (EBV)-positive cancer-specific tumor antigen epitope, represented by any one of SEQ ID NOs: 1 to 3, SEQ ID NOs: 7 to 9, SEQ ID NOs: 11 to 15, SEQ ID NOs: 19 to 21, SEQ ID NOs: 23 to 39, SEQ ID NOs: 43 to 45, SEQ ID NOs: 47 to 51, SEQ ID NOs: 55 to 57, SEQ ID NOs: 59 to 63, SEQ ID NOs: 67 to 69, SEQ ID NOs: 71 to 75, SEQ ID NOs: 79 to 81, SEQ ID NOs: 83 to 111, SEQ ID NOs: 115 to 117, SEQ ID NOs: 119 to 122, SEQ ID NOs: 124 to 127, SEQ ID NOs: 129 to 149, SEQ ID NOs: 151 to 154, SEQ ID NOs: 156 to 158, SEQ ID NOs: 160 to 163, SEQ ID NOs: 165 to 167, SEQ ID NOs: 168 to 176, SEQ ID NOs: 178 to 181, SEQ ID Nos: 183 to 194, and SEQ ID NOs: 196 to 199.

[0097] According to another embodiment of the present invention, there is provided an antigen-presenting cell (APC) loaded with a cancer-specific tumor antigen epitope provided in the present invention.

[0098] According to yet another embodiment of the present invention, there is provided a T cell activated by an antigen-presenting cell provided in the present invention.

[0099] According to still yet another embodiment of the present invention, there is provided an anticancer vaccine or a pharmaceutical composition for preventing or treating cancer, comprising, as an active ingredient, an antigen-presenting cell loaded with a cancer-specific tumor antigen epitope provided in the present invention; and/or an activated T cell.

[0100] According to still yet another embodiment of the present invention, there is provided a method for preventing or treating cancer, comprising administering, to an individual, an effective amount of an antigen-presenting cell loaded with a cancer-specific tumor antigen epitope provided in the present invention; and/or an activated T cell.

[0101] Hereinafter, the present invention will be described in more detail by way of examples. These examples are only for describing the present invention in more detail, and it will be apparent to those skilled in the art that according to the gist of the present invention, the scope of the present invention is not limited by these examples.

EXAMPLES

[0102] [Example 1] Method for Producing EBV-Positive Gastric Cancer Cell Antigen Epitopes

[0103] Prediction of epitopes was made such that the epitope has 15-mer amino acids in the entire sequence of LMP2a or EBNA-1 as shown in Tables 1 and 2; and the epitopes in the sequence listing, which are set forth in Tables 1 (LMP2a) and 2 (EBNA-1), were produced according to a conventional method related to peptide production processes.

TABLE-US-00001 TABLE 1 SEQ ID NO Amino acid sequence SEQ ID NO: 1 MGSLEMVPMGAGPPS SEQ ID NO: 2 EMVPMGAGPPSPGGD SEQ ID NO: 3 MGAGPPSPGGDPDGD SEQ ID NO: 4 PPSPGGDPDGDDGGN SEQ ID NO: 5 GGDPDGDDGGNNSQY SEQ ID NO: 6 DGDDGGNNSQYPSAS SEQ ID NO: 7 GGNNSQYPSASGSYG SEQ ID NO: 8 SQYPSASGSYGNTPT SEQ ID NO: 9 SASGSYGNTPTPPND SEQ ID NO: 10 SYGNTPTPPNDEERE SEQ ID NO: 11 TPTPPNDEERESNEE SEQ ID NO: 12 PNDEERESNEEPPPP SEQ ID NO: 13 ERESNEEPPPPYEDL SEQ ID NO: 14 NEEPPPPYEDLDWGN SEQ ID NO: 15 PPPYEDLDWGNGDRH SEQ ID NO: 16 EDLDWGNGDRHSDYQ SEQ ID NO: 17 WGNGDRHSDYQPLGN SEQ ID NO: 18 DRHSDYQPLGNQDPS SEQ ID NO: 19 DYQPLGNQDPSLYLG SEQ ID NO: 20 LGNQDPSLYLGLQHD SEQ ID NO: 21 DPSLYLGLQHDGNDG SEQ ID NO: 22 YLGLQHDGNDGLPPP SEQ ID NO: 23 QHDGNDGLPPPPYSP SEQ ID NO: 24 NDGLPPPPYSPRDDS SEQ ID NO: 25 PPPPYSPRDDSSQHI SEQ ID NO: 26 YSPRDDSSQHIYEEA SEQ ID NO: 27 DDSSQHIYEEAGRGS SEQ ID NO: 28 QHIYEEAGRGSMNPV SEQ ID NO: 29 EEAGRGSMNPVCLPV SEQ ID NO: 30 RGSMNPVCLPVIVAP SEQ ID NO: 31 NPVCLPVIVAPYLFW SEQ ID NO: 32 LPVIVAPYLFWLAAI SEQ ID NO: 33 VAPYLFWLAAIAASC SEQ ID NO: 34 LFWLAAIAASCFTAS SEQ ID NO: 35 AAIAASCFTASVSTV SEQ ID NO: 36 ASCFTASVSTVVTAT SEQ ID NO: 37 TASVSTVVTATGLAL SEQ ID NO: 38 STVVTATGLALSLLL SEQ ID NO: 39 TATGLALSLLLLAAV SEQ ID NO: 40 LALSLLLLAAVASSY SEQ ID NO: 41 LLLLAAVASSYAAAQ SEQ ID NO: 42 AAVASSYAAAQRKLL SEQ ID NO: 43 SSYAAAQRKLLTPVT SEQ ID NO: 44 AAQRKLLTPVTVLTA SEQ ID NO: 45 KLLTPVTVLTAVVTF SEQ ID NO: 46 PVTVLTAVVTFFAIC SEQ ID NO: 47 LTAVVTFFAICLTWR SEQ ID NO: 48 VTFFAICLTWRIEDP SEQ ID NO: 49 AICLTWRIEDPPFNS SEQ ID NO: 50 TWRIEDPPFNSLLFA SEQ ID NO: 51 EDPPFNSLLFALLAA SEQ ID NO: 52 FNSLLFALLAAAGGL SEQ ID NO: 53 LFALLAAAGGLQGIY SEQ ID NO: 54 LAAAGGLQGIYVLVM SEQ ID NO: 55 GGLQGIYVLVMLVLL SEQ ID NO: 56 GIYVLVMLVLLILAY SEQ ID NO: 57 LVMLVLLILAYRRRW SEQ ID NO: 58 VLLILAYRRRWRRLT SEQ ID NO: 59 LAYRRRWRRLTVCGG SEQ ID NO: 60 RRWRRLTVCGGIMFL SEQ ID NO: 61 RLTVCGGIMFLACVL SEQ ID NO: 62 CGGIMFLACVLVLIV SEQ ID NO: 63 MFLACVLVLIVDAVL SEQ ID NO: 64 CVLVLIVDAVLQLSP SEQ ID NO: 65 LIVDAVLQLSPLLGA SEQ ID NO: 66 AVLQLSPLLGAVTVV SEQ ID NO: 67 LSPLLGAVTVVSMTL SEQ ID NO: 68 LGAVTVVSMTLLLLA SEQ ID NO: 69 TVVSMTLLLLAFVLW SEQ ID NO: 70 MTLLLLAFVLWLSSP SEQ ID NO: 71 LLAFVLWLSSPGGLG SEQ ID NO: 72 VLWLSSPGGLGTLGA SEQ ID NO: 73 SSPGGLGTLGAALLT SEQ ID NO: 74 GLGTLGAALLTLAAA SEQ ID NO: 75 LGAALLTLAAALALL SEQ ID NO: 76 LLTLAAALALLASLI SEQ ID NO: 77 AAALALLASLILGTL SEQ ID NO: 78 ALLASLILGTLNLTT SEQ ID NO: 79 SLILGTLNLTTMFLL SEQ ID NO: 80 GTLNLTTMFLLMLLW SEQ ID NO: 81 LTTMFLLMLLWTLVV SEQ ID NO: 82 FLLMLLWTLVVLLIC SEQ ID NO: 83 LLWTLVVLLICSSCS SEQ ID NO: 84 LVVLLICSSCSSCPL SEQ ID NO: 85 LICSSCSSCPLTKIL SEQ ID NO: 86 SCSSCPLTKILLARL SEQ ID NO: 87 CPLTKILLARLFLYA SEQ ID NO: 88 KILLARLFLYALALL SEQ ID NO: 89 ARLFLYALALLLLAS SEQ ID NO: 90 LYALALLLLASALIA SEQ ID NO: 91 ALLLLASALIAGGSI SEQ ID NO: 92 LASALIAGGSILQTN SEQ ID NO: 93 LIAGGSILQTNFKSL SEQ ID NO: 94 GSILQTNFKSLSSTE SEQ ID NO: 95 QTNFKSLSSTEFIPN SEQ ID NO: 96 KSLSSTEFIPNLFCM SEQ ID NO: 97 STEFIPNLFCMLLLI SEQ ID NO: 98 IPNLFCMLLLIVAGI SEQ ID NO: 99 FCMLLLIVAGILFIL SEQ ID NO: 100 LLIVAGILFILAILT SEQ ID NO: 101 AGILFILAILTEWGS SEQ ID NO: 102 FILAILTEWGSGNRT SEQ ID NO: 103 ILTEWGSGNRTYGPV SEQ ID NO: 104 WGSGNRTYGPVFMCL SEQ ID NO: 105 NRTYGPVFMCLGGLL SEQ ID NO: 106 GPVFMCLGGLLTMVA SEQ ID NO: 107 MCLGGLLTMVAGAVW SEQ ID NO: 108 GLLTMVAGAVWLTVM SEQ ID NO: 109 MVAGAVWLTVMTNTL SEQ ID NO: 110 AVWLTVMTNTLLSAW SEQ ID NO: 111 TVMTNTLLSAWILTA SEQ ID NO: 112 NTLLSAWILTAGFLI SEQ ID NO: 113 SAWILTAGFLIFLIG SEQ ID NO: 114 LTAGFLIFLIGFALF SEQ ID NO: 115 FLIFLIGFALFGVIR SEQ ID NO: 116 LIGFALFGVIRCCRY SEQ ID NO: 117 ALFGVIRCCRYCCYY SEQ ID NO: 118 VIRCCRYCCYYCLTL SEQ ID NO: 119 CRYCCYYCLTLESEE SEQ ID NO: 120 CYYCLTLESEERPPT SEQ ID NO: 121 LTLESEERPPTPYRN SEQ ID NO: 122 LESEERPPTPYRNTV

TABLE-US-00002 TABLE 2 SEQ ID NO Amino acid sequence SEQ ID NO: 123 GGAGAGGGGRGRGGS SEQ ID NO: 124 AGGGGRGRGGSGGRG SEQ ID NO: 125 GRGRGGSGGRGRGGS SEQ ID NO: 126 GGSGGRGRGGSGGRG SEQ ID NO: 127 GRGRGGSGGRGRGGS SEQ ID NO: 128 GGSGGRGRGGSGGRR SEQ ID NO: 129 GRGRGGSGGRRGRGR SEQ ID NO: 130 GGSGGRRGRGRERAR SEQ ID NO: 131 GRRGRGRERARGGSR SEQ ID NO: 132 RGRERARGGSRERAR SEQ ID NO: 133 RARGGSRERARGRGR SEQ ID NO: 134 GSRERARGRGRGRGE SEQ ID NO: 135 RARGRGRGRGEKRPR SEQ ID NO: 136 RGRGRGEKRPRSPSS SEQ ID NO: 137 RGEKRPRSPSSQSSS SEQ ID NO: 138 RPRSPSSQSSSSGSP SEQ ID NO: 139 PSSQSSSSGSPPRRP SEQ ID NO: 140 SSSSGSPPRRPPPGR SEQ ID NO: 141 GSPPRRPPPGRRPFF SEQ ID NO: 142 RRPPPGRRPFFHPVG SEQ ID NO: 143 PGRRPFFHPVGDADY SEQ ID NO: 144 PFFHPVGDADYFEYL SEQ ID NO: 145 PVGDADYFEYLQEGG SEQ ID NO: 146 ADYFEYLQEGGPDGE SEQ ID NO: 147 EYLQEGGPDGEPDVP SEQ ID NO: 148 EGGPDGEPDVPPGAI SEQ ID NO: 149 DGEPDVPPGAIEQGP SEQ ID NO: 150 DVPPGAIEQGPTDDP SEQ ID NO: 151 GAIEQGPTDDPGEGP SEQ ID NO: 152 QGPTDDPGEGPSTGP SEQ ID NO: 153 DDPGEGPSTGPRGQG SEQ ID NO: 154 EGPSTGPRGQGDGGR SEQ ID NO: 155 TGPRGQGDGGRRKKG SEQ ID NO: 156 GQGDGGRRKKGGWFG SEQ ID NO: 157 GGRRKKGGWFGKHRG SEQ ID NO: 158 KKGGWFGKHRGQGGS SEQ ID NO: 159 WFGKHRGQGGSNPKF SEQ ID NO: 160 HRGQGGSNPKFENIA SEQ ID NO: 161 GGSNPKFENIAEGLR SEQ ID NO: 162 PKFENIAEGLRVLLA SEQ ID NO: 163 NIAEGLRVLLARSHV SEQ ID NO: 164 GLRVLLARSHVERTT SEQ ID NO: 165 LLARSHVERTTEEGN SEQ ID NO: 166 SHVERTTEEGNWVAG SEQ ID NO: 167 RTTEEGNWVAGVFVY SEQ ID NO: 168 EGNWVAGVFVYGGSK SEQ ID NO: 169 VAGVFVYGGSKTSLY SEQ ID NO: 170 FVYGGSKTSLYNLRR SEQ ID NO: 171 GSKTSLYNLRRGIAL SEQ ID NO: 172 SLYNLRRGIALAVPQ SEQ ID NO: 173 LRRGIALAVPQCRIT SEQ ID NO: 174 IALAVPQCRITPLSR SEQ ID NO: 175 VPQCRITPLSRLPFG SEQ ID NO: 176 RITPLSRLPFGMAPG SEQ ID NO: 177 LSRLPFGMAPGPGPQ SEQ ID NO: 178 PFGMAPGPGPQPGPL SEQ ID NO: 179 APGPGPQPGPLRESI SEQ ID NO: 180 GPQPGPLRESIVCYF SEQ ID NO: 181 GPLRESIVCYFMVFL SEQ ID NO: 182 ESIVCYFMVFLQTHI SEQ ID NO: 183 CYFMVFLQTHIFAEV SEQ ID NO: 184 VFLQTHIFAEVLKDA SEQ ID NO: 185 THIFAEVLKDAIKDL SEQ ID NO: 186 AEVLKDAIKDLVMTK SEQ ID NO: 187 KDAIKDLVMTKPAPT SEQ ID NO: 188 KDLVMTKPAPTCNIK SEQ ID NO: 189 MTKPAPTCNIKVTVC SEQ ID NO: 190 APTCNIKVTVCSFDD SEQ ID NO: 191 NIKVTVCSFDDGVDL SEQ ID NO: 192 TVCSFDDGVDLPPWF SEQ ID NO: 193 FDDGVDLPPWFPPMV SEQ ID NO: 194 VDLPPWFPPMVEGAA SEQ ID NO: 195 PWFPPMVEGAAAEGD SEQ ID NO: 196 PMVEGAAAEGDDGDD SEQ ID NO: 197 GAAAEGDDGDDGDEG SEQ ID NO: 198 EGDDGDDGDEGGDGD SEQ ID NO: 199 GDDGDEGGDGDEGEE SEQ ID NO: 200 GDEGGDGDEGEEGQE

[Example 2] ELISPOT Results for T Cells Activated by Dendritic Cells Loaded with Selected LMP2a Epitope

[0104] PBMCs extracted from the blood of three healthy humans (N5, N9, or N15) were separated into monocytes and leukocytes through magnetic-activated cell sorting, and the monocytes were cultured for 4 days in a culture supplemented with cytokines GM-CSF and IL-4 to differentiate into dendritic cells. To the culture, in which the monocytes had differentiated into dendritic cells, was added each group containing 10 or 11 epitope peptides as set forth in Table 3 together with poly(I:C) so that the respective peptides were transferred to the dendritic cells. The culture was matured for 1 day. In addition, the leukocytes were cultured for 3 days in a culture supplemented with anti-CD3/CD28 antibodies and cytokine IL-2.

TABLE-US-00003 TABLE 3 Group SEQ ID NO 1 SEQ ID NO: 1, SEQ ID NO: 13, SEQ ID NO: 25, SEQ ID NO: 37, SEQ ID NO: 49, SEQ ID NO: 61, SEQ ID NO: 73, SEQ ID NO: 85, SEQ ID NO: 97, SEQ ID NO: 109, SEQ ID NO: 121 2 SEQ ID NO: 2, SEQ ID NO: 14, SEQ ID NO: 26, SEQ ID NO: 38, SEQ ID NO: 50, SEQ ID NO: 62, SEQ ID NO: 74, SEQ ID NO: 86, SEQ ID NO: 98, SEQ ID NO: 110, SEQ ID NO: 122 3 SEQ ID NO: 3, SEQ ID NO: 15, SEQ ID NO: 27, SEQ ID NO: 39, SEQ ID NO: 51, SEQ ID NO: 63, SEQ ID NO: 75, SEQ ID NO: 87, SEQ ID NO: 99, SEQ ID NO: 111 4 SEQ ID NO: 4, SEQ ID NO: 16, SEQ ID NO: 28, SEQ ID NO: 40, SEQ ID NO: 52, SEQ ID NO: 64, SEQ ID NO: 76, SEQ ID NO: 88, SEQ ID NO: 100, SEQ ID NO: 112 5 SEQ ID NO: 5, SEQ ID NO: 17, SEQ ID NO: 29, SEQ ID NO: 41, SEQ ID NO: 53, SEQ ID NO: 65, SEQ ID NO: 77, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 113 6 SEQ ID NO: 6, SEQ ID NO: 18, SEQ ID NO: 30, SEQ ID NO: 42, SEQ ID NO: 54, SEQ ID NO: 66, SEQ ID NO: 78, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 114 7 SEQ ID NO: 7, SEQ ID NO: 19, SEQ ID NO: 31, SEQ ID NO: 43, SEQ ID NO: 55, SEQ ID NO: 67, SEQ ID NO: 79, SEQ ID NO: 91, SEQ ID NO: 103, SEQ ID NO: 115 8 SEQ ID NO: 8, SEQ ID NO: 20, SEQ ID NO: 32, SEQ ID NO: 44, SEQ ID NO: 56, SEQ ID NO: 68, SEQ ID NO: 80, SEQ ID NO: 92, SEQ ID NO: 104, SEQ ID NO: 116 9 SEQ ID NO: 9, SEQ ID NO: 21, SEQ ID NO: 33, SEQ ID NO: 45, SEQ ID NO: 57, SEQ ID NO: 69, SEQ ID NO: 81, SEQ ID NO: 93, SEQ ID NO: 105, SEQ ID NO: 117 10 SEQ ID NO: 10, SEQ ID NO: 22, SEQ ID NO: 34, SEQ ID NO: 46, SEQ ID NO: 58, SEQ ID NO: 70, SEQ ID NO: 82, SEQ ID NO: 94, SEQ ID NO: 106, SEQ ID NO: 118 11 SEQ ID NO: 11, SEQ ID NO: 23, SEQ ID NO: 35, SEQ ID NO: 47, SEQ ID NO: 59, SEQ ID NO: 71, SEQ ID NO: 83, SEQ ID NO: 95, SEQ ID NO: 107, SEQ ID NO: 119 12 SEQ ID NO: 12, SEQ ID NO: 24, SEQ ID NO: 36, SEQ ID NO: 48, SEQ ID NO: 60, SEQ ID NO: 72, SEQ ID NO: 84, SEQ ID NO: 96, SEQ ID NO: 108, SEQ ID NO: 120 13 SEQ ID NO: 1 to SEQ ID NO: 12 14 SEQ ID NO: 13 to SEQ ID NO: 24 15 SEQ ID NO: 25 to SEQ ID NO: 36 16 SEQ ID NO: 37 to SEQ ID NO: 48 17 SEQ ID NO: 49 to SEQ ID NO: 60 18 SEQ ID NO: 61 to SEQ ID NO: 72 19 SEQ ID NO: 73 to SEQ ID NO: 84 20 SEQ ID NO: 85 to SEQ ID NO: 96 21 SEQ ID NO: 97 to SEQ ID NO: 108 22 SEQ ID NO: 109 to SEQ ID NO: 120 23 SEQ ID NO: 121 and SEQ ID NO: 122

[0105] Then, CD8-expressing T cells were isolated from leukocytes, which were cultured in a culture supplemented with anti-CD3/CD28 antibody and cytokine IL-2, by magnetic-activated cell sorting, and co-cultured with mature dendritic cells at a ratio of 1:10 (dendritic cells:CD8-expressing T cells). In a case of the co-culture, the culture was mixed with a cytokine cocktail containing cytokines IL-4 and IL-7 that function to help survival and immune response of T cells, and the mixture was cultured. After 18 hours, secretion levels of IFN-γ in the T cells thus activated were measured with ELISPOT, and the results are illustrated in FIGS. 1 to 4. Here, as the positive control peptide, a peptide consisting of the sequence of 58.sup.th to 66.sup.th amino acids in the M1 protein, which is one of the proteins expressed by influenza virus, was employed.

[0106] As illustrated in FIGS. 1 and 2, for N5 (N5-1 and N5-2; the same experiment was repeated twice using the same person's blood), it was identified that the dendritic cells transformed with the epitope group 3, 7, 9, 11, 12, 15, 18, or 21 resulted in increased expression of IFN-γ in T cells.

[0107] As illustrated in FIG. 3, for N9, it was identified that the dendritic cells transformed with the epitope group 1, 2, 3, 7, 8, 10, 15, 16, 19, or 20 resulted in increased expression of IFN-γ in T cells.

[0108] In addition, as illustrated in FIG. 4, for N15, it was identified that the dendritic cells transformed with the epitope group 1, 2, 3, 5, 6, 7, 8, 9, 11, 15, 21, or 22 resulted in increased expression of IFN-γ in T cells.

[0109] From these results, it was found that in the present invention, cytotoxic T lymphocytes (CTLs) can be activated by dendritic cells loaded with epitopes selected from the epitopes as set forth in Tables 1 and 2 above, and the thus activated T cells have antigen specificity which enables recognition of the epitope that is a novel antigen.

[Example 3] ELISPOT Results for T Cells Activated with Dendritic Cells Loaded with Selected EBNA-1 Epitope

[0110] Using PBMCs extracted from the blood of three healthy humans (N2, N15, or N19), the degree of T cell activation was checked in the same manner as in Example 2, and the results are illustrated in FIGS. 5 to 8. Here, the monocytes having differentiated into dendritic cells were transformed with each group containing 10 or 11 epitope peptides as set forth in Table 4.

TABLE-US-00004 TABLE 4 Group SEQ ID NO  1′ SEQ ID NO: 123, SEQ ID NO: 132, SEQ ID NO: 141, SEQ ID NO: 150, SEQ ID NO: 159, SEQ ID NO: 168, SEQ ID NO: 177, SEQ ID NO: 186, SEQ ID NO: 195  2′ SEQ ID NO: 124, SEQ ID NO: 133, SEQ ID NO: 142, SEQ ID NO: 151, SEQ ID NO: 160, SEQ ID NO: 169, SEQ ID NO: 178, SEQ ID NO: 187, SEQ ID NO: 196  3′ SEQ ID NO: 125, SEQ ID NO: 134, SEQ ID NO: 143, SEQ ID NO: 152, SEQ ID NO: 161, SEQ ID NO: 170, SEQ ID NO: 179, SEQ ID NO: 188, SEQ ID NO: 197  4′ SEQ ID NO: 126, SEQ ID NO: 135, SEQ ID NO: 144, SEQ ID NO: 153, SEQ ID NO: 162, SEQ ID NO: 171, SEQ ID NO: 180, SEQ ID NO: 189, SEQ ID NO: 198  5′ SEQ ID NO: 127, SEQ ID NO: 136, SEQ ID NO: 145, SEQ ID NO: 154, SEQ ID NO: 163, SEQ ID NO: 172, SEQ ID NO: 181, SEQ ID NO: 190, SEQ ID NO: 199  6′ SEQ ID NO: 128, SEQ ID NO: 137, SEQ ID NO: 146, SEQ ID NO: 155, SEQ ID NO: 164, SEQ ID NO: 173, SEQ ID NO: 182, SEQ ID NO: 191, SEQ ID NO: 200  7′ SEQ ID NO: 129, SEQ ID NO: 138, SEQ ID NO: 147, SEQ ID NO: 156, SEQ ID NO: 165, SEQ ID NO: 174, SEQ ID NO: 183, SEQ ID NO: 192  8′ SEQ ID NO: 130, SEQ ID NO: 139, SEQ ID NO: 148, SEQ ID NO: 157, SEQ ID NO: 166, SEQ ID NO: 175, SEQ ID NO: 184, SEQ ID NO: 193  9′ SEQ ID NO: 131, SEQ ID NO: 140, SEQ ID NO: 149, SEQ ID NO: 158, SEQ ID NO: 167, SEQ ID NO: 176, SEQ ID NO: 185, SEQ ID NO: 194 10′ SEQ ID NO: 123 to SEQ ID NO: 131 11′ SEQ ID NO: 132 to SEQ ID NO: 140 12′ SEQ ID NO: 141 to SEQ ID NO: 149 13′ SEQ ID NO: 150 to SEQ ID NO: 158 14′ SEQ ID NO: 159 to SEQ ID NO: 167 15′ SEQ ID NO: 168 to SEQ ID NO: 176 16′ SEQ ID NO: 177 to SEQ ID NO: 185 17′ SEQ ID NO: 186 to SEQ ID NO: 194 18′ SEQ ID NO: 195 to SEQ ID NO: 200

[0111] As illustrated in FIG. 5, for N2, it was identified that cytotoxic T lymphocytes were activated by the dendritic cells transformed with the epitope group 6′, 7′, 8′, or 15′.

[0112] In addition, as illustrated in FIGS. 6 and 7, for N15 (N15-1 and N15-2; the same experiment was repeated twice using the same person's blood), it was identified that cytotoxic T lymphocytes were activated by the dendritic cells transformed with each of all epitope groups except for some groups.

[0113] In addition, as illustrated in FIG. 8, for N19, it was identified that cytotoxic T lymphocytes were activated by the dendritic cells transformed with the epitope group 3′, 4′, 5′, 9′, 12′, or 15′.

[0114] From the results of Examples 2 and 3, it can be seen that the epitopes according to the present invention can very effectively activate cytotoxic T lymphocytes.

[0115] Although specific parts of the present invention have been described in detail, it is obvious to those skilled in the art that such a specific description is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

INDUSTRIAL APPLICABILITY

[0116] The present invention relates to a cancer-specific tumor antigen epitope, an antigen-presenting cell loaded with the epitope, and a method for activating T cells for cancer treatment by the antigen-presenting cell.