HTLV-1中和エピトープの同定および抗HTLV-1ペプチドワクチンの開発

• 研究課題/領域番号: 04670391
• 研究種目: 一般研究(C)
• 配分区分: 補助金
• 研究分野: 内科学一般
• 研究機関: 九州大学
• 研究代表者: 中村 稔 九州大学, 医学部, 助手 (40217906)
• 研究分担者: 中野 修治 九州大学, 医学部, 助手 (40164248) ; 永渕 正法 九州大学, 医学部, 助手 (00150441)
• 研究期間 (年度): 1992 – 1993
• 研究課題ステータス: 完了 (1993年度)
• 配分額: 2,100千円 (直接経費: 2,100千円); 1993年度: 800千円 (直接経費: 800千円); 1992年度: 1,300千円 (直接経費: 1,300千円)
• キーワード: Bリンパ球レパートリー / ヒト型モノクローナル抗体 / 抗HTLV-1抗体 / 中和エピトープ / ペプチドワクチン / B cellエピトープ / T-cellエピトープ / 抗ミトコンドリア抗体 / 抗HTLV-1 抗体 / B Cell エピトープ / T Cell エピトープ / 抗HTLV-I抗体 / B cell エピトープ / T-cell エピトープ

研究概要

ヒト型の抗HTLV-1モノクローナル抗体(mAb)を用いて、HTLV-1の中和エピトープを詳細に解析した後、HTLV-1ペプチドワクチンの作製が可能か否かについて検討した。まず、HAM/TSP患者末梢血Bリンパ球にEpstein Barr virus(EBV)を感染させ、HTLV-1 envelope gl-ycoprotein(gp46)と結合する抗HTLV-1 mAbを多数作製した。153種のgp46と結合するmAbの内、24種が中和活性を有していた。これらの中和mAbの内、13種がgp46 peptide 175-199と反応し、1種がgp46peptide 213-236に,他の1種がgp46 peptide 288-317と結合した。9種のmAbは結合するpeptideが同定できなかった。また、gp46 pepti-de 175-199と結合するmAbを用いて、4つの異なる中和エピトープ;(1)187-193,(2)191-191,(3)193-199,(4)continuous conformat-ional B cell epitope,をその領域内に同定し、gp46 a.a.187-199をprinciple neutralizing determinant(PND)と命名した。液相のペプチドを用いた競合実験により、PND natural gp46 proteinの中和エピトープと極めて類似した構造をとることから、ペプチドワクチンの良い候補と考えられた。そこで、PNDを含む様々なペプチドワクチンを合成し、ラットあるいはラビットに免疫し、高力価の中和抗体を誘導することに成功した(J.Immunol in press,1994).今後、HTLV-1 T cell epitopeの解析をさらにすすめ、より効率良く中和抗体を誘導するペプチドワクチンの開発を目指す予定である。

研究成果

• [文献書誌] Eishi Baba and Minoru Nakamura et.al: "A peptide based HTLV-1 vaccine containing T and B cell epitopes which induces high titers of neutralizing antibodies." The Journal of Immunology. (in press). (1994)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Minoru Nakamura et al: "Peripheral B lymphocyte repertoire to mitochondrial antigens in patients with primary biliary cirrhosis.-positive correlation between the disease activity and the freguency of circulating B lymphocytes committed to the production of IyG to AMA." Hepatology. (in press). (1994)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Jun-ichi Kira et al: "Sequence heterogeneity of HTLV-1 proviral DNA in the central nervous system of patients with HTLV-1 associated myelopathy." Annuals of Neurology. (in press). (1994)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Hideyuki Ikematsu et al: "VH and VK segment structure of anti-Insulin IyG autoantibodies in patients with lnsulin-dependent diabetes mellitus.Evidence for somatic mutation." The Journal of Immunology. (in press). (1994)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Mieko Matsui and Minoru Nakamura et al: "Human monoclonal autoantibodies from a patient with primary blliary cirrhosis which recognize two distinct autoepitoges in the E2 component of the pyruvate dehydrogenase complex." Hepatology. 18. 1069-1077 (1993)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Eishi Baba and Minoru Nakamura et al: "Multiple neutralizing B cell epitopes of HTLV-1 identified by human monoclonal antibodies.-A basis for the design of an HTLV-1 peptide vaccine-" The Journal of Immunology. 151. 1013-1024 (1993)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Mika Kuroki and Minoru Nakamura et al: "Identification of new epitopes recognized by human monoclonal antibodies with neutrallzing and antibody-dependent cellular cytotoxicity activities specific for HTLV-1." The Journal of Immunology. 149. 940-948 (1992)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Minoru Nakamura and Paolo Casali: "Generation of human monoclonal autoantibody-producing cell lines by Epstein-Barr virus(EBV)-transformation of autoreactive B lymphocytes followed by somatic cell hybridization." Immunomethods,Acodemic Press,Inc.San Diego.USA, 159-168 (1992)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Eishi Baba and Minoru Nakamura et al: "A peptide based HTLV-1 vaccine containing T and B cell epitopes which induce high titers of neutralizing antibodies." J.Immunol. (in press). (1994)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Minoru Nakamura et al: "Peripheral B lymphocyte repertoire to mitochondrial antigens in patients with primary biliary cirrhosis." Hepatology. (in press). (1944)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Jun-ichi Kira et al: "Sequence heterogeneity of HTLV-1 proviral DNA in the central nervous system of patients with HTLV-1 associated myelopathy." Annuals of Nneurology. (in press). (1994)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Hideyuki Ikematsu et al: "VH and VK segment structure of anti-insulin IgG autoantibodies in patients with insulin-dependent diabetes mellitus." J.Immunol. (in press). (1994)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Mieko Matsui and Minoru Nakamura et al: "Human monoclonal autoantibodies in a patient with primary biliary cirrhosis which recognize two distinct autoepietopes in the E2 component of the pyruvate dehydrogenase complex." Hepatology. 18. 1069-1077 (1993)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Eishi Baba and Minoru Nakamura et al: "Multilpe neutralizing B cell epotopes of HTLV-1 identified by human monoclonal antibodies. - A basis of the design of an HTLV-1 peptide vaccine -" J.Immunol. 151. 1013-1024 (1993)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Mika Kuroki and Minoru Nakamura et al: "Identification of new epitopes recognized by human monoclonal antibodies with neutralizing and antibody-dependent cellular cytotoxicity activities specific for HTLV-1." J.Immunol. 149. 940-948 (1992)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Minoru Nakamura et al: "Increase of peripheral B lymphocytes committed to the production of monoreactive and high affinity antibodies to HLTV-1 in patients with HAM/TSP." J.Neuroimmunol.37. 35-45 (1992)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Minoru Nakamura and Paolo Casali: "Generation of human monoclonal autoantibody-producing cell lines by Epstein Barr virus-transformation of autoreactive B lymphocytes followed by somatic cell hybridization." Immunomethods. 1. 159-168 (1992)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Eishi,Baba and Minoru,Nakamura et al.: "A peptide based HTLV-1 vaccine containing T and B cell epitopes which induces high titers of neutralizing antibodies." The Journal of Immunology. (in press). (1994)
• [文献書誌] Minoru,Nakamura et al.: "Peripheral B lymphocyte repertoire to mitochondrial antigens in patients with primery biliary cirrhosis.-Positive correlation between the disease activity and the frecuency of ." Hepatology. (in press). (1994)
• [文献書誌] Jun-ichi,Kira et al.: "Sequence heterogeneity of HTLV-1 proviral DNA in the central nervous system of patients with HTLV-1 associated myelopathy." Annuals of Neurology. (in press). (1994)
• [文献書誌] Hideyuki,Ikematsu et al.: "VH and VK segment structure of anti-Insulim Ig G autoantibodies in patients with insulin-dependent diabetesmellitus.Evidence for somatic mutation.21GC04:The Journal of Immunology." (in press). (1994)
• [文献書誌] Mieko,Matsui and Minoru,Nakamura et al.: "Human monocloxal autoantibodies apatient with primary biliary cirrhosiswhich recognize two distinct autoepitopes in the E2 component of the pyruvatedehydrogenase complex." Hepatology. 18. 1069-1077 (1993)
• [文献書誌] Eishi,Baba and Minoru,Nakamura et al.: "Multiple neutralizing B cell epitopes of HTLV-1 identified by human monoclonal antibodies.-A basis for the design of an HTLV-1 peptide vaccine-." The Journal of Immunology.151. 1013-1024 (1993)
• [文献書誌] Mika,Kuroki and Minoru,Nakamuraet al.: "Identification of new epitopes recognized by human monoclonal antibodies with neutralizing and antibody-dependent cellular cytotoxicity activities specific for HTLV-1." The Journal of Immunology.149. 940-948 (1992)
• [文献書誌] Minoru,Nakamura and Paoro,Casali.: "Generation of human monoclonal autoantibedy-producing cell lines by Epstein-Barr virus(EVB)-transformation ofautoreactive B lymphocytes followed bysomatic cell hybridization." Immunomethods,Academic Press,Inc.San Diego U.S.A., 159-168 (1992)
• [文献書誌] Minoru Nakamura: "Elevated antibodies to synthetic peptides of HTLV-I envelope transmembrane glycoproteins with HAM/TSP." Journal of Neuroimmunology. 35. 167-177 (1991)
• [文献書誌] Minoru Nakamura: "Increase of peripheral B lymphocytes committed to the production of monoreactive-and high affinty-antibodies to HTLV-I in patients with HAM/TSP." Journal of Neuroimmunology. 37. 35-45 (1992)
• [文献書誌] Jun-ichi Kira: "Antibodies titers to HTLV-I p40 tax protein and gag-env hybrid protein in HTLV-I-associated myelopathy/tropical spastic paraparesis:correlation with increased HTLV-I proviral DNA load." Journal of the Neurological Sciences. 107. 98-104 (1992)
• [文献書誌] Mika-Kuroki: "Identification of new epitopes recognized by human monoclonal antibodies with neutrolizing and antibody-dependent cellular cytotoxicity activities specific for human T-cell leukemia virus type I." The Journal of Immunology. 149. 940-948 (1992)
• [文献書誌] Mika-Kuroki: "In vivo comparative the rapeutic study of optimal adminitration of 5-fluorouracil and cisplatin using a newly establised HST-I human squamous-carcinoma cell line." Cancer Chemother Pharmacol. 29. 273-276 (1992)
• [文献書誌] Taito Esaki: "Inhibition by 5-fluorouracil of cis-diamminedichloroplatinum(II)-induced DNA interstrand cross-link removal in a HST-I humam squamous carcinoma cell line" Cancer Research. 52. 6501-6506 (1992)
• [文献書誌] Kenji Mitsugi: "Simple and efficient chemosensitivity assay for human primary cells using contact-sensitive microtiter plates" International Journal of oncology. 1. 141-147 (1992)
• [文献書誌] Minoru Nakamura: "Generation of human monoclonal autoantibody-producing cell lines by Epstein-Barr virus(EBV)-transformation of autoreactive B lymphocytes followed by somatic cell hybridization." Immunomethods,Academic Press,Inc.San Diego,U.S.A., IN PRESS