Identification of human T-cell epitopes on Mycobacterium tuberculosis small molecular weight proteins and development of DNA vaccines based on the epitopes

• Project/Area Number: 16390130
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bacteriology (including Mycology)
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: NAGATA Toshi Hamamatsu Univ. Sch. Med., Professor, 医学部, 教授 (90275024)
• Co-Investigator(Kenkyū-buntansha): KOIDE Yukio Hamamatsu Univ. Sch. Med., Professor, 医学部, 教授 (30126809) ; UCHIJIMA Masato Hamamatsu Univ. Sch. Med., Research associate, 医学部, 助手 (20252174) ; 青枝 大貴 浜松医科大学, 医学部, 助手 (10324344)
• Project Period (FY): 2004 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥14,000,000 (Direct Cost: ¥14,000,000); Fiscal Year 2006: ¥3,000,000 (Direct Cost: ¥3,000,000); Fiscal Year 2005: ¥3,000,000 (Direct Cost: ¥3,000,000); Fiscal Year 2004: ¥8,000,000 (Direct Cost: ¥8,000,000)
• Keywords: DNA vaccine / tuberculosis / MPT51 / CTL epitope / HLA transgenic mice / Thエピトープ / HLA-Aトランスジェニックマウス

Research Abstract

CD8^+ cytotoxic T lymphocytes (CTL) play a pivotal role in protection against Mycobacterium tuberculosis infection. We identified a novel CTL epitope on a protective antigen of M. tuberculosis, MPT51, in HLA-A^*0201-transgenic HHD mice. HHD mice were immunized with plasmid DNA encoding MPT51 using gene gun and interferon (IFN)-y production from the immune splenocytes was analyzed. In response to synthetic overlapping peptide library covering the mature MPT51 sequence, only one peptide, p51-70, stimulated the splenocytes to produce IFN-y. Three-color flow cytometric analysis of cell-surface CD4 and CD8 staining revealed that p51-70 peptide contains an immunodominant CD8^+ T-cell epitope. Further analysis using computer-assisted algorithms permitted identification of a bona fide T-cell epitope, p53-62. MHC class I stabilization assay using T2 cells confirmed that the epitope is presented by HLA-A^*0201. MPT51 p53-62/HLA-A^*0201 tetramer was observed to bind to the specific CD8^+ T cells of immune HHD splenocytes. In addition, MPT51 p53-62-specific memory CD8^+ T cells were found in tuberculin skin test-positive HLA-A^*0201^+ healthy individuals. This HLA-A^*0201-restricted CD8^+ T-cell epitope would be feasible for analysis of the role of MPT51-specific T cells in protective immunity and for future vaccine design against M. tuberculosis infection.

Research Products

• [Journal Article] Immunization with dendritic cells retrovirally transduced with mycobacterial antigen 85A gene elicits the specific cellular immunity including cytotoxic T-lymphocyte activity specific to a dominant epitope on antigen 85A. (2006)
  • Author(s): Nakano, H., Nagata, T., Suda, T., Tanaka, T., Aoshi, T., Uchijima, M., Chida, K., Nakamura, H., Okada, M., Koide, Y.
  • Journal Title: Vaccine 24 Pages: 2110-2119
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Immunization with gene encoding granulocyte-macrophage colony-stimulating factor inserted with a single helper T-cell epitope of an intracellular bacterium induces a specific T-cell subset and protective immunity. (2006)
  • Author(s): Nagata, T., Uchijima, M., Uchiyama, H., Yamada, T., Aoshi, T., Koide, Y.
  • Journal Title: Vaccine 24 Pages: 4538-4553
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Curcumin maintenance therapy for ulcerative colitis : Randomized multicenter double blind placebo-controlled clinical trial. (2006)
  • Author(s): Hanai, H., Iida, T., Takeuchi, K., Watanabe, F., Maruyama, Y., Andoh, A., Tsujikawa, T., Fujiyama, Y., Mitsuyama, K., Sata, M., Yamada, M., Iwaoka, Y., Kanke, K., Hiraishi, H., Hirayama, K., Arai, H., Yoshii, S., Uchijima, M., Nagata, T., Koide, Y.
  • Journal Title: Clin. Gastroenterol. Hepatol. 4 Pages: 1502-1506
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Immunization with dendritic cells pulsesd with α-galactocylceramide and a dominant CTL epitope elicits effective protective immunity against intracellular bacteria infection. (2006)
  • Author(s): Enomoto, N., Nagata, T., Suda, T., Uchijima, M., Chida, K., Nakamura, H., Koide, Y.
  • Journal Title: Int. J. Infect. Dis. 10
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Intratracheal administration of third-generation lentivirus vector encoding Mpt51 from M. tuberculosis induces lung-homing specific T cells. (2006)
  • Author(s): Koide, Y., Hashimoto, D., Uchijima, M., Suda, T., Chida, K., Nagata, T.
  • Journal Title: Int. J. Infect. Dis. 10
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Curcumin maintenance therapy for ulcerative colitis : Randomized multicenter double blind placebo-controlled clinical trial. (2006)
  • Author(s): Hanai, H., Iida, T., Takeuchi, K., Watanabe, F., Maruyama, Y., Andoh, A., Tsujikawa, T., Fujiyama, Y., Mitsuyama, K., Sata, M., Yamada, M., Iwaoka, Y., Kanke, K., Hiraishi, H., Hirayama, K., Arai, H., Yoshii, S., Uchijima, M., Nagata, T., Koide, Y.
  • Journal Title: Clinical Gastroenterol. Hepatol. 4 Pages: 1502-1506
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Immunization with dendritic cells pulsesd with a-galactocylceramide and a dominant CTL epitope elicits effective protective immunity against intracellular bacteria infection. (2006)
  • Author(s): Enomoto, N., Nagata, T., Suda, T., Uchijima, M., Chida, K., Nakamura, H., Koide, Y.
  • Journal Title: Int. J. Infect. Dis. 10
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Intratracheal administration of third-generation lentivirus vector encoding Mpt51 from M tuberculosis induces lung-homing specific T cells. (2006)
  • Author(s): Koide, Y., Hashimoto, D., Uchijima, M., Suda, T., Chida, K., Nagata, T.
  • Journal Title: Int. J. Infect. Dis. 10
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Immunization with gene encoding granulocyte-macrophage colony-stimulating factor inserted with a single helper T-cell epitope of an intracellular bacterium induces a specific T-cell subset and protective immunity. (2006)
  • Author(s): Nagata T., Uchijima, M., Uchiyama, H., Yamada, T., Aoshi, T., Koide, Y.
  • Journal Title: Vaccine 24 Pages: 4538-4553
• [Journal Article] Curcumin Maintenance Therapy for Ulcerative Colitis : Randomized Multicenter Double Blind Placebo-Controlled Clinical Trial. (2006)
  • Author(s): Hanai, H., Iida, T., Takeuchi, K., Watanabe, F., Maruyama, Y., Andoh, A., Tsujikawa, T., Fujiyama, Y., Mitsuyama, K., Sata, M., Yamada, M., Iwaoka, Y., Kanke, K., Hiraishi, H., Hirayama, K., Arai, H., Yoshii, S., Uchijima, M., Nagata, T., Koide, Y.
  • Journal Title: Clinical Gastroenterology and Hepatology 4・12 Pages: 1502-1506
• [Journal Article] Immunization with gene encoding granulocyte-macrophage colony-stimulating factor inserted with a single helper T-cell epitope of an intracellular bacterium induces a specific T-cell subset and protective immunity. (2006)
  • Author(s): Nagata, T., Uchijima, M., Uchiyama, H., Yamada, T., Aoshi, T., Koide, Y.
  • Journal Title: Vaccine (in press)
• [Journal Article] Interferon-γ overcomes low responsiveness of myeloid dendritic cells to CpG-DNA. (2005)
  • Author(s): Uchijima, M., Nagata, T., Aoshi, T., Koide, Y.
  • Journal Title: Immunol. Cell Biol. 83 Pages: 92-95
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Expression mapping by retroviral vector for CD8+ T cell epitopes : definition of a Mycobacterium tuberculosis peptide presented by H2-D^d. (2005)
  • Author(s): Aoshi, T., Suzuki, M., Uchijima, M., Nagata, T., Koide, Y.
  • Journal Title: J. Immunol. Methods 298 Pages: 21-34
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Effects of oxygen radicals in low-pressure surface-wave plasma on sterilization. (2005)
  • Author(s): Nagatsu, M., Terashita, F., Nonaka, H., Xu, L., Nagata, T., Koide, Y.
  • Journal Title: Appl. Phys. Letters 86
  • NAID: 120000796467
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Interferon-y overcomes low responsiveness of myeloid dendritic cells to CpG-DNA. (2005)
  • Author(s): Uchijima, M., Nagata, T., Aoshi, T., Koide, Y.
  • Journal Title: Immunol. Cell Biol. 83 Pages: 92-95
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Expression mapping by retroviral vector for CD8+ T cell epitopes : definition of a Mycobacterium tuberculosis peptide presented by H2-Dd. (2005)
  • Author(s): Aoshi, T., Suzuki, M., Uchijima, M., Nagata, T., Koide, Y.
  • Journal Title: J. Immunol. Methods. 298 Pages: 21-34
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Effects of oxygen radicals in low-pressure surface-wave plasma on sterilization. (2005)
  • Author(s): Nagatsu, M., Terashita, F., Nonaka, H., Xu, L., Nagata, T., Koide, Y.
  • Journal Title: Appl. Phys. Letters. 86
  • NAID: 120000796467
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Interferon-γ overcomes low responsiveness of myeloid dendritic cells to CpG-DNA. (2005)
  • Author(s): Uchijima M, Nagata T, Aoshi T, Koide Y.
  • Journal Title: Immunolgy and Cell Biology 83 Pages: 92-95
• [Journal Article] Expression mapping by retroviral vector for CD8^+ T cell epitopes : definition of a Mycobacterium tuberculosis peptide presented by H2-D^d. (2005)
  • Author(s): Aoshi T, Suzuki M, Uchijima M, Nagata T, Koide Y.
  • Journal Title: Journal of Immunological Methods 298 Pages: 21-34
• [Journal Article] Effects of oxygen radicals in low-pressure surface-wave plasma on sterilization. (2005)
  • Author(s): Nagatsu, M., Terashita, F., Nonaka, H., Xu, L., Nagata.T., Koide, Y.
  • Journal Title: Applied Physical Letters 86
  • NAID: 120000796467
• [Journal Article] Interferon-y overcomes low responsiveness of myeloid dendritic cells to CpG-DNA. (2005)
  • Author(s): Uchijima M, Nagata T, Aoshi T, Koide Y.
  • Journal Title: Immunolgy and Cell Biology 83 Pages: 92-95
• [Journal Article] Expression mapping by retroviral vector for CD8^+ T cell epitopes : definition of a Mycobacterium tuberculosis peptide presented by H2-D^d. (2005)
  • Author(s): Aoshi T, Suzuki M, Uchijima M, Nagata T, Koide Y.
  • Journal Title: Journal of Immunological Methods (in press)
• [Journal Article] Induction of protective cellular immunity against Mycobacterium tuberculosis by recombinant attenuated self-destructing Listeria monocytogenes strains harboring eukaryotic expression plasmids for Ag85 complex and MPB/MPT51. (2004)
  • Author(s): Miki, K., Nagata, T., Tanaka, T., Kim, Y.-H., Uchijima, M., Ohara, N., Nakamura, S., Okada, M., Koide, Y.
  • Journal Title: Infect. Immun. 72・4 Pages: 2014-2021
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of MHC-restricted T-cell epitopes on a novel protective antigen, MPT51, of Mycobacterium tuberculosis. (2004)
  • Author(s): Koide, Y., Suzuki, M., Aoshi, T., Nagata, T.
  • Journal Title: Int. J. Infect. Dis. 8
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Plasmid encoding interleukin-4 in the amelioration of murine collagen-induced arthritis. (2004)
  • Author(s): Kageyama, Y., Koide, Y., Uchijima, M., Nagata, T., Yoshida, A., Aoshi, T., Miura, T., Nagafusa, T., Nagano, A.
  • Journal Title: Arthritis Rhumat. 50 Pages: 968-975
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of H2-D^d- and H2-A^b-restricted T-cell epitopes on a novel protective antigen, MPT51, of Mycobacterium tuberculosis. (2004)
  • Author(s): Suzuki, M., Aoshi, T., Nagata, T., Koide, Y.
  • Journal Title: Infect. Immun. 72・7 Pages: 3829-3837
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Butyrate suppresses hypoxia-inducible factor-1 activity in intestinal epithelial cells under hypoxic conditions. (2004)
  • Author(s): Miki, K., Unno, N., Nagata, T., Uchijima, M., Konno, H., Koide, Y., Nakamura, S.
  • Journal Title: Shock 22・5 Pages: 446-452
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] α-Galactosylceramide, a ligand of natural killer T cells, inhibits allergic airway inflammation. (2004)
  • Author(s): Matsuda, H., Suda, T., Sato, J., Nagata, T., Koide, Y., Chida, K., Nakamura, H.
  • Journal Title: Am. J. Respir. Cell. Mol. Biol. 33・1 Pages: 22-31
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cytokine and antibody production during the course of resolution in Plasmodium yoelii 17XL-infected BALB/c mice treated with febrifugine and isofebrifugine mixture from leaves of Hydrangea macrophylla var. Otaksa. (2004)
  • Author(s): Ishih, A., Nagata, T., Kobayashi, F., Miyase, T., Terada, M.
  • Journal Title: Parasitol. Res. 94・3 Pages: 176-182
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cytotoxic T-lymphocyte-, and helper T-lymphocyte-oriented DNA vaccine. (2004)
  • Author(s): Nagata, T., Aoshi, T., Uchijima, M., Suzuki, M., Koide, Y.
  • Journal Title: DNA Cell Biol. 23・2 Pages: 93-106
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Attenuated bacteria as transfer vehicles of DNA vaccines. (2004)
  • Author(s): Nagata, T., Miki, K., Koide, Y.
  • Journal Title: Recent Res. Devel. Biophys. Biochem. 4 Pages: 189-207
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Induction of protective cellular immunity against Mycobacterium tuberculosis by recombinant attenuated self-destructing Listeria monocytogenes strains harboring eukaryotic expression plasmids for Ag85 complex and MPB/MPT51. (2004)
  • Author(s): Miki, K., Nagata, T., Tanaka, T., Kim, Y.-H., Uchijima, M., Ohara, N., Nakamura, S., Okada, M., Koide, Y.
  • Journal Title: Infect. Immun. 72(4) Pages: 2014-2021
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Plasmid encoding interleukin-4 in the amelioration of murine collagen-induced arthritis. (2004)
  • Author(s): Kageyama, Y., Koide, Y., Uchijima, M., Nagata, T., Yoshida, A., Aoshi, T., Miura, T., Nagafusa, T., Nagano, A.
  • Journal Title: Arthritis Rheumat. 50 Pages: 968-975
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification of H2-Dd-and H2-Ab-restricted T-cell epitopes on a novel protective antigen, MPT51, of Mycobacterium tuberculosis. (2004)
  • Author(s): Suzuki, M., Aoshi, T., Nagata, T., Koide, Y.
  • Journal Title: Infect. Immun. 72(7) Pages: 3829-3837
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Butyrate suppresses hypoxia-inducible factor-1 activity in intestinal epithelial cells under hypoxic conditions. (2004)
  • Author(s): Miki, K., Unno, N., Nagata, T., Uchijima, M., Konno, H., Koide, Y., Nakamura, S.
  • Journal Title: Shock 22(5) Pages: 446-452
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] a-Galactosylceramide, a ligand of natural killer T cells, inhibits allergic airway inflammation. (2004)
  • Author(s): Matsuda, H., Suda, T., Sato, J., Nagata, T., Koide, Y., Chida, K., Nakamura, H.
  • Journal Title: Am. J. Respir. Cell. Mol. Biol. 33(1) Pages: 22-31
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cytokine and antibody production during the course of resolution in Plasmodium yoelii 17XL-infected BALB/c mice treated with febrifugine and isofebrifugine mixture from leaves of Hydrangea macrophylla var. Otaksa. (2004)
  • Author(s): Ishih, A., Nagata, T., Kobayashi, F., Miyase, T., Terada, M.
  • Journal Title: Prasitol. Res. 94(3) Pages: 176-182
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cytotoxic T-lymphocyte-, and helper T-lymphocyte-oriented DNA vaccine. (2004)
  • Author(s): Nagata, T., Aoshi, T., Uchijima, M., Suzuki, M., Koide, Y.
  • Journal Title: DNA Cell Biol. 23(2) Pages: 93-106
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Induction of protective immunity against Mycobacterium tuberculosis by recombinant attenuated self-destructing Listeria monocytogenes strains harboring eukaryotic expression plasmids for Ae85 complex and MPB/MPT51. (2004)
  • Author(s): Miki K, Nagata T, Tanaka T, Kim Y-H, Uchijima, M, Ohara N, Nakamura S.
  • Journal Title: Infection and Immunity 72(4) Pages: 2014-2021
• [Journal Article] Plasmid encoding interleukin-4 in the amelioration of murine collagen-induced arthritis. (2004)
  • Author(s): Kageyama Y, Koide Y, Uchijima M, Nagata T, Yoshida A, Aoshi T, Miura T, Nagafusa T, Nagano A.
  • Journal Title: Arthritis and Rheumatitis 50 Pages: 968-975
• [Journal Article] Identification of H2-D^d- and H2-A^d-restricted T-cell epitopes on a novel protective antigen, MPT51, of Mycobacterium tuberculosis. (2004)
  • Author(s): Suzuki M, Aoshi T, Nagata T, Koide Y.
  • Journal Title: Infection and Immunity 72(7) Pages: 3829-3837
• [Journal Article] Cytotoxic T-lymphocyte-, and helper T-lymphocyte-oriented DNA vaccine. (2004)
  • Author(s): Nagata T, Aoshi T, Uchijima M, Suzuki M, Koide Y.
  • Journal Title: DNA and Cell Biology 23 Pages: 93-106
• [Journal Article] Butyrate suppresses hypoxia-inducible factor-1 activity in intestinal epithelial cells under hypoxic conditions. (2004)
  • Author(s): Milk K, Unno N, Nagata T, Uchijima M Konno H, Koide Y, Nakamura S.
  • Journal Title: Shock 22 Pages: 446-452
• [Journal Article] Attenuated bacteria as transfer vehicles of DNA vaccines. (2004)
  • Author(s): Nagata T, Miki K, Koide Y.
  • Journal Title: Recent Research Development of Biophysics and Biochemstry 4 Pages: 189-207