2003 年度実績報告書

ヘリコバクター・ピロリのウレアーゼ活性を抑制する抗体の認識エピトープの同定

研究課題

研究課題/領域番号	13670481
研究機関	日本医科大学
研究代表者	高橋秀実日本医科大学, 大学院・医学研究科, 教授 (40221361)
キーワード	Helicobacter pylori / urease / 中和抗体 / epitope / MAP法 / mapping / vaccine / 酵素活性
研究概要	これまで申請者らは、Helicobacter pylori(以下Hpと略す)が胃粘膜に結合し定着するための鍵を握る菌体表面に位置する酵素であるurease、ことにそのlarge subunitであるB鎖が患者血清抗体の主たる認識抗原であることを見出した(日本消化器病学会誌、91巻、2202-2213、1994)。その際、胃炎の表層性変化にはureaseに対するIgA抗体の上昇が、また萎縮性変化にはIgG抗体の上昇が関連するすることを見出すとともに(Gut、43:168-175,1998)、Hp-ureaseに対するIgG抗体には病状進行を促進するものと、Hpの感染を予防するものとがあることを想定した。そこで、本研究では、まず精製Hp-urease抗原をBALB/cマウスに投与し、その免疫脾臓細胞からHp-urease特異的なhybridomaを作成し、その中よりHp-ureaseの酵素活性(尿素分解能)を抑制するものを見出し、合成ペプチド群を用いてその認識部分のエピトープの同定を試みた。本科学研究費の補助により申請者らは、Hpのurease、ことにその酵素活性を抑制する中和抗体の最小エピトープをB鎖内の8個のアミノ酸からなるペプチドF8 (SIKEDVQF)として同定すること、そしてその中でも329番目のアミノ酸であるリジン(K)がHp-ureaseの活性特性の鍵を握ることを見いだし、その内容を国際誌(Infect.Immun.69:6597-6603,2001)に掲載した。また、このF8部分は同時期に報告されたHp-ureaseの酵素活性の活性中心(Nat.Struct.Biol.8:505-509,2001)と完全に一致していることを確認した。現在この中和エピトープを用いたHp感染予防および除菌効力のあるワクチンの開発を目指している。

研究成果
(27件)

すべてその他

すべて文献書誌 (27件)

[文献書誌] Takahashi, M., Nakagawa, Y., Berzofsky, J.A., Takahashi, H.: "Counter-regulation of 1cytolytic activity and cytokine production in human immunodeficiency virus (HIV)-1-specific murine CD8+ CTL free antigenic peptide."Int.Immunol.. 13. 43-51 (2001)
[文献書誌] Nakajima, Y., Takahashi, M., Norose, Y., Ogawa, R., Takahashi, H.: "Induction of apoptosis in mice thymocytes by tetracaine."Biomed.Res.. 21. 297-303 (2001)
[文献書誌] Watari, E., Shinya, E., Kurane, S., Takahashi, H.: "Effect of cyclosporin A on cell fusion in monkey kidney cell line persistently infected with measles virus."Intervirol.. 301. 209-211 (2001)
[文献書誌] Machida, K., Takahashi, H., et al.: "Inhibition of cytochrome C release in Fas-mediated signaling pathway by hepatitis C viral proteins in transgenic mice liver."J.Biol.Chem.. 276. 12140-12146 (2001)
[文献書誌] Hirota, K., Takahashi, H., et al.: "Identification of an antigenic epitope in Helicobacter pylori urease that induces neutralizing antibody production."Infect.Immu.. 69. 6597-6603 (2001)
[文献書誌] Fujimoto, C., Nakagawa, Y., Shimizu, M., Ohara, K., Takahashi, H.: "Isolation of antigenic substances from HIV-1 envelope gp120 gene transfectants by mild acid elution and X-iirradiation treatment : for the development of CTL-based---"Biomed.Res.. 24. 115-124 (2003)
[文献書誌] Hamada, H., Hiroi, T., Nishiyama, Y., Takahashi, H., et al.: "Identification of multiple isolated lymphoid follicles on the antimesenteric wall of the mouse small intestine."J.Immunol.. 168. 57-64 (2002)
[文献書誌] Nishiyama, Y., Takahashi, H., et al.: "Homeostatic regulation of intestinal villous epithelia by B lymphocytes."J.Immunol.. 168. 2626-2633 (2002)
[文献書誌] Yokosuka, T., Takahashi, H., et al.: "Predominant role of T cell receptor (TCR)-α chain in forming preimmune TCR repertoire revieled by clonal TCR reconstitution system."J.Exp.Med.. 195. 991-1001 (2002)
[文献書誌] Enose, Y., Takahashi, H., et al.: "Protection by intranasal immunization of a nef-deleted, nonpathogenic SHIV against intravaginal challenge with a heterologous pathogenic SHIV."Virology. 298. 306-316 (2002)
[文献書誌] Li, Q., Nagahara, N., Takahashi, H., Takeda, K., Okumura, K., Minami, M.: "Organophosphorus pesticides markedly inhibit the activities of natural killer, cytotoxic T lymphocyte and lymphokine-activated killer : a proposed inhibiting ---"Toxicology. 172. 181-190 (2002)
[文献書誌] Takahashi, M., Osono, E., Nakagawa, Y., Takahashi., H.: "Rapid induction of apoptosis in CD8^+ HIV-1 envelope-specific murine CTLs short exposure to antigenic peptide."J.Immunol.. 169. 6588-6593 (2002)
[文献書誌] Ichikawa, M., Takahashi, H., et al.: "Brest milk macrophages spontaneously produce GM-CSF and differentiate into dendritic cells in the presence of exogenous IL-4 alone."Immunology. 108. 189-195 (2003)
[文献書誌] Sakaue, G., Takahashi, H., et al.: "HIV Mucosal Vaccine : Nasal Immunization with gp160-Encapsulated Hemagglutinating Virus of Japan-Liposome Induces Antigen-Specific CTLs ---"J.Immunol.. 170. 495-502 (2003)
[文献書誌] Takaku, S., Takahashi, H., et al.: "Induction of hepatic injury by hepatitic C virus-specific CD8+ murine cytotoxic T lymphocytes in transgenic mice expressing the viral structural genes."Biochem.Biophys.Res.Commun.. 301. 330-337 (2003)
[文献書誌] Hidemi Takahashi: "Antigen presentation in vaccine development."Comp.Immunol.Microbiol.& Infect.Dis.. 26. 309-328 (2003)
[文献書誌] Takeuchi, J., Takahashi, H., et al.: "Down-regulation of Toll-like receptor expression in monocyte-derived Langerhans cell-like cells : implications of low-responsiveness to bacterial components in the ---"Biochem.Biophys.Res.Commun.. 306. 674-679 (2003)
[文献書誌] Kawashima, T., Takahashi, H., Brenner, M.B., Sugita, M., et al.: "Major CD8 T cell response to live bacillus Calmette-Guerin is madiated by CD1 molecules."J.Immunol.. 170. 5345-5348 (2003)
[文献書誌] Shinya, E., Takahashi, H., et al.: "Effect of Nef-deleted pseudotyeped HIV virions bearing an enhanced green fluorescent protein gene in the env on HIV-sensitive transformed T cells."Biomed.Res.. 24. 59-69 (2003)
[文献書誌] Akahata, W., Takahashi, H., et al.: "DNA vaccination of macaques by a full-genome simian/human immunodeficiency virus type 1 plasmid chimera that produces non-infectious virus particles."J.Gen.Virol.. 84. 2237-2244 (2003)
[文献書誌] Kim Y., Takahashi, H., et al.: "Induction of CD4+ murine natural killer T-like cells by immunization with syngeneic thymoma expressing embryonic α-fetoprotein."Cell.Immunol.. 226. 1-10 (2003)
[文献書誌] Yanagihara, T., Takahashi, H., et al.: "Age dependent decrease of polymeric Ig receptor expression and IgA elevation in ddY mice : a possible cause of IgA nephropathy."Lab.Invest.. 84. 63-70 (2004)
[文献書誌] Fujimoto, C., Nakagawa, Y., Ohara, K., Takahashi, H.: "Polyriboinosinic polyribocytidylic acid (Poly(I : C))/TLR3 signaling allows class I processing of exogenous protein and induction of HIV-specific CD8+ CTLs."Int.Immunol.. 16. 55-63 (2004)
[文献書誌] Ishii, R., Shimizu, M., Nakagawa, Y., Shimizu, K., Tanaka, S., Takahashi, H: "In vivo priming of natural killer T cells by dendritic cells pulsed with hepatoma-derived acid-eluted substance."Cancer Immunol.Immunother.. 53. 383-390 (2004)
[文献書誌] 高橋秀実(分担): "免疫学イラストレイティッド"南江堂. 424 (2001)
[文献書誌] 高橋秀実(分担): "分子生物学・免疫学キーワード辞典"医学書院. 1035 (2003)
[文献書誌] 高橋秀実(監修): "ブラック微生物学"丸善出版. 866 (2003)

2003 年度 実績報告書

ヘリコバクター・ピロリのウレアーゼ活性を抑制する抗体の認識エピトープの同定

研究代表者

高橋 秀実 日本医科大学, 大学院・医学研究科, 教授 (40221361)

研究成果

[文献書誌] Takahashi, M., Nakagawa, Y., Berzofsky, J.A., Takahashi, H.: "Counter-regulation of 1cytolytic activity and cytokine production in human immunodeficiency virus (HIV)-1-specific murine CD8+ CTL free antigenic peptide."Int.Immunol.. 13. 43-51 (2001)

[文献書誌] Nakajima, Y., Takahashi, M., Norose, Y., Ogawa, R., Takahashi, H.: "Induction of apoptosis in mice thymocytes by tetracaine."Biomed.Res.. 21. 297-303 (2001)

[文献書誌] Watari, E., Shinya, E., Kurane, S., Takahashi, H.: "Effect of cyclosporin A on cell fusion in monkey kidney cell line persistently infected with measles virus."Intervirol.. 301. 209-211 (2001)

[文献書誌] Machida, K., Takahashi, H., et al.: "Inhibition of cytochrome C release in Fas-mediated signaling pathway by hepatitis C viral proteins in transgenic mice liver."J.Biol.Chem.. 276. 12140-12146 (2001)

[文献書誌] Hirota, K., Takahashi, H., et al.: "Identification of an antigenic epitope in Helicobacter pylori urease that induces neutralizing antibody production."Infect.Immu.. 69. 6597-6603 (2001)

[文献書誌] Fujimoto, C., Nakagawa, Y., Shimizu, M., Ohara, K., Takahashi, H.: "Isolation of antigenic substances from HIV-1 envelope gp120 gene transfectants by mild acid elution and X-iirradiation treatment : for the development of CTL-based---"Biomed.Res.. 24. 115-124 (2003)

[文献書誌] Hamada, H., Hiroi, T., Nishiyama, Y., Takahashi, H., et al.: "Identification of multiple isolated lymphoid follicles on the antimesenteric wall of the mouse small intestine."J.Immunol.. 168. 57-64 (2002)

[文献書誌] Nishiyama, Y., Takahashi, H., et al.: "Homeostatic regulation of intestinal villous epithelia by B lymphocytes."J.Immunol.. 168. 2626-2633 (2002)

[文献書誌] Yokosuka, T., Takahashi, H., et al.: "Predominant role of T cell receptor (TCR)-α chain in forming preimmune TCR repertoire revieled by clonal TCR reconstitution system."J.Exp.Med.. 195. 991-1001 (2002)

[文献書誌] Enose, Y., Takahashi, H., et al.: "Protection by intranasal immunization of a nef-deleted, nonpathogenic SHIV against intravaginal challenge with a heterologous pathogenic SHIV."Virology. 298. 306-316 (2002)

[文献書誌] Li, Q., Nagahara, N., Takahashi, H., Takeda, K., Okumura, K., Minami, M.: "Organophosphorus pesticides markedly inhibit the activities of natural killer, cytotoxic T lymphocyte and lymphokine-activated killer : a proposed inhibiting ---"Toxicology. 172. 181-190 (2002)

[文献書誌] Takahashi, M., Osono, E., Nakagawa, Y., Takahashi., H.: "Rapid induction of apoptosis in CD8^+ HIV-1 envelope-specific murine CTLs short exposure to antigenic peptide."J.Immunol.. 169. 6588-6593 (2002)

[文献書誌] Ichikawa, M., Takahashi, H., et al.: "Brest milk macrophages spontaneously produce GM-CSF and differentiate into dendritic cells in the presence of exogenous IL-4 alone."Immunology. 108. 189-195 (2003)

[文献書誌] Sakaue, G., Takahashi, H., et al.: "HIV Mucosal Vaccine : Nasal Immunization with gp160-Encapsulated Hemagglutinating Virus of Japan-Liposome Induces Antigen-Specific CTLs ---"J.Immunol.. 170. 495-502 (2003)

[文献書誌] Takaku, S., Takahashi, H., et al.: "Induction of hepatic injury by hepatitic C virus-specific CD8+ murine cytotoxic T lymphocytes in transgenic mice expressing the viral structural genes."Biochem.Biophys.Res.Commun.. 301. 330-337 (2003)

[文献書誌] Hidemi Takahashi: "Antigen presentation in vaccine development."Comp.Immunol.Microbiol.& Infect.Dis.. 26. 309-328 (2003)

[文献書誌] Takeuchi, J., Takahashi, H., et al.: "Down-regulation of Toll-like receptor expression in monocyte-derived Langerhans cell-like cells : implications of low-responsiveness to bacterial components in the ---"Biochem.Biophys.Res.Commun.. 306. 674-679 (2003)

[文献書誌] Kawashima, T., Takahashi, H., Brenner, M.B., Sugita, M., et al.: "Major CD8 T cell response to live bacillus Calmette-Guerin is madiated by CD1 molecules."J.Immunol.. 170. 5345-5348 (2003)

[文献書誌] Shinya, E., Takahashi, H., et al.: "Effect of Nef-deleted pseudotyeped HIV virions bearing an enhanced green fluorescent protein gene in the env on HIV-sensitive transformed T cells."Biomed.Res.. 24. 59-69 (2003)

[文献書誌] Akahata, W., Takahashi, H., et al.: "DNA vaccination of macaques by a full-genome simian/human immunodeficiency virus type 1 plasmid chimera that produces non-infectious virus particles."J.Gen.Virol.. 84. 2237-2244 (2003)

[文献書誌] Kim Y., Takahashi, H., et al.: "Induction of CD4+ murine natural killer T-like cells by immunization with syngeneic thymoma expressing embryonic α-fetoprotein."Cell.Immunol.. 226. 1-10 (2003)

[文献書誌] Yanagihara, T., Takahashi, H., et al.: "Age dependent decrease of polymeric Ig receptor expression and IgA elevation in ddY mice : a possible cause of IgA nephropathy."Lab.Invest.. 84. 63-70 (2004)

[文献書誌] Fujimoto, C., Nakagawa, Y., Ohara, K., Takahashi, H.: "Polyriboinosinic polyribocytidylic acid (Poly(I : C))/TLR3 signaling allows class I processing of exogenous protein and induction of HIV-specific CD8+ CTLs."Int.Immunol.. 16. 55-63 (2004)

[文献書誌] Ishii, R., Shimizu, M., Nakagawa, Y., Shimizu, K., Tanaka, S., Takahashi, H: "In vivo priming of natural killer T cells by dendritic cells pulsed with hepatoma-derived acid-eluted substance."Cancer Immunol.Immunother.. 53. 383-390 (2004)

[文献書誌] 高橋秀実(分担): "免疫学イラストレイティッド"南江堂. 424 (2001)

[文献書誌] 高橋秀実(分担): "分子生物学・免疫学キーワード辞典"医学書院. 1035 (2003)

[文献書誌] 高橋秀実(監修): "ブラック微生物学"丸善出版. 866 (2003)

2003 年度実績報告書

高橋秀実日本医科大学, 大学院・医学研究科, 教授 (40221361)