1994 Fiscal Year Annual Research Report

ビブリオ属菌の産生するいわゆる溶血毒の病原的意義の解析

Research Project

Project/Area Number	04454196
Research Institution	Osaka University
Principal Investigator	本田武司大阪大学, 微生物病研究所, 教授 (60029808)
Co-Investigator(Kenkyū-buntansha)	余明順大阪大学, 微生物病研究所, 助手 (70093482) 山本耕一郎大阪大学, 微生物病研究所, 助教授 (30158274)
Keywords	腸炎ビブリオ / コレラ菌 / 耐熱性溶血毒 / エルトール型ヘモリシン / リン酸化
Research Abstract	代表的なビブリオ属菌の産生する溶血毒として、(1)腸炎ビブリオの耐熱性溶血毒(TDH)及びその類似毒素(TRH)と(2)コレラ菌の産生するエルトール型溶血毒の二つを取り上げこれらについて解析した。 1.腸炎ビブリオのTDH/TRH類似毒素に関する成績 (1)TDHに類似した新奇な溶血毒として見い出したTRH産生菌をしらべたところ、TRH産生性とurease産生性に相関性があることを見出した。 (2)TDHとTRHを同時に産生している菌株を見い出し、これを蛋白及び遺伝子側から実証した。 (3)TDH/TRHは膜上に存在するレセプター(これは非感受性細胞にもあることも明らかにした)に結合後、プロテインキナーゼCを介して25kDa膜蛋白がリン酸化される結果、外径約30nm、内径1-2nm大のporeを形成し、細胞破壊がおこることを明らかにした。 (4)TRH遺伝子を欠失した腸炎ビブリオ変異株を作成し、病原性を調べたところ、病原性は減弱するものの、完全には消失しなかった。この事からTRHが病原因子の一つで有る事を示すとともにTDH/TRH以外の病原因子の存在も示唆された。 2.コレラ菌のエルトール型溶血毒の解析 (1)エルトール型溶血毒は80kDaのpre-protoxinとして産生され、signal peptideが切断されprotoxin(79kDa)となり、この分子中のpro領域が「分子内シャペロン」として外膜通過を促し分泌されること。 (2)分泌されたprotoxinは34kDaのHA/proteaseにより活性型(成熟型)溶血毒となること。 (3)コレラ菌はエルトール型溶血毒以外にこれまで知られていない新奇な溶血毒遺伝子を持つこと、などを明らかにした。

Research Products

(12 results)

All Other

All Publications (12 results)

[Publications] Xu,M.et al.: "Demonstration and characterization of simultaneous production of a TDH and a TRH by a clinical isolate of V.parahaemolyticus" Infection and Immunity. 62. 166-171 (1994)
[Publications] Tang,G.Q.et al.: "A mutant toxin of V.parahaemlyticus thermostable direct hemolysin which has lost hemolytic activity but retains ability to bind" Infection and Immunity. 62. 3299-3304 (1994)
[Publications] Xu,M.et al.: "Construction and characterization of an isogenic mutant of V.parahaemolyticus having a deletion inthe TRH gene" Journal of Bacteriology. 176. 4754-4760 (1994)
[Publications] Taniguchi,T.et al.: "Evidence that the N-terminal amino acid sequence of pilus colonization factor antigen III produced by human ETEC is similar to" Journal of Infectious Diseases. 170. 1049-1050 (1994)
[Publications] Ichinise,Y. et al.: "The effect on enterotoxicityof protease purified from Vibrio cholerae 01." FEMS Microbiol Letters. 115. 265-272 (1994)
[Publications] Miyagi,K. et al.: "Successful application of enzyme-1 beled oligonucleotide probe for rapid and accurate cholera-diagnosis in a clinical diagnosis" Microbiology and Immunology. 38. 301-304 (1994)
[Publications] Nagayama,K. et al.: "Correlation between cell-associated mannose-sensitive hemagglutination by V.parahaemolyticus and adherence to a human colonic" FEMS Microbiology Letters. 120. 207-210 (1994)
[Publications] Shimaoka,M. et al.: "Development of enzyme-labeled oligonucleotide probe for mecA gene in MRSA." Journal of Clincal Microbiology. 32. 1866-1869 (1994)
[Publications] Iijima,Y. et al.: "Simultaneous outbreak due to Vibrio cholerae and Shigella dysenteriae in Kenya." Lancet. 345. 69-70 (1995)
[Publications] Yoh,M. et al.: "Evaluation of two assay kits for TDH as an indicator of TRH produced by V.parahaemolyticus." Microbiology and Immunology. 39. 157-159 (1995)
[Publications] Honda,T. et al.: "A novel method to chemically immobilize antibodyon nylon and its application to the rapid and diferential detection of two toxins" Clinical and Diagnostic Laboratory Immunology. 2(in press). (1995)
[Publications] 本田,武司: "病気と細菌毒素" 化学同人, 159 (1994)

1994 Fiscal Year Annual Research Report

ビブリオ属菌の産生するいわゆる溶血毒の病原的意義の解析

Principal Investigator

本田 武司 大阪大学, 微生物病研究所, 教授 (60029808)

Research Products

[Publications] Xu,M.et al.: "Demonstration and characterization of simultaneous production of a TDH and a TRH by a clinical isolate of V.parahaemolyticus" Infection and Immunity. 62. 166-171 (1994)

[Publications] Tang,G.Q.et al.: "A mutant toxin of V.parahaemlyticus thermostable direct hemolysin which has lost hemolytic activity but retains ability to bind" Infection and Immunity. 62. 3299-3304 (1994)

[Publications] Xu,M.et al.: "Construction and characterization of an isogenic mutant of V.parahaemolyticus having a deletion inthe TRH gene" Journal of Bacteriology. 176. 4754-4760 (1994)

[Publications] Taniguchi,T.et al.: "Evidence that the N-terminal amino acid sequence of pilus colonization factor antigen III produced by human ETEC is similar to" Journal of Infectious Diseases. 170. 1049-1050 (1994)

[Publications] Ichinise,Y. et al.: "The effect on enterotoxicityof protease purified from Vibrio cholerae 01." FEMS Microbiol Letters. 115. 265-272 (1994)

[Publications] Miyagi,K. et al.: "Successful application of enzyme-1 beled oligonucleotide probe for rapid and accurate cholera-diagnosis in a clinical diagnosis" Microbiology and Immunology. 38. 301-304 (1994)

[Publications] Nagayama,K. et al.: "Correlation between cell-associated mannose-sensitive hemagglutination by V.parahaemolyticus and adherence to a human colonic" FEMS Microbiology Letters. 120. 207-210 (1994)

[Publications] Shimaoka,M. et al.: "Development of enzyme-labeled oligonucleotide probe for mecA gene in MRSA." Journal of Clincal Microbiology. 32. 1866-1869 (1994)

[Publications] Iijima,Y. et al.: "Simultaneous outbreak due to Vibrio cholerae and Shigella dysenteriae in Kenya." Lancet. 345. 69-70 (1995)

[Publications] Yoh,M. et al.: "Evaluation of two assay kits for TDH as an indicator of TRH produced by V.parahaemolyticus." Microbiology and Immunology. 39. 157-159 (1995)

[Publications] Honda,T. et al.: "A novel method to chemically immobilize antibodyon nylon and its application to the rapid and diferential detection of two toxins" Clinical and Diagnostic Laboratory Immunology. 2(in press). (1995)

[Publications] 本田,武司: "病気と細菌毒素" 化学同人, 159 (1994)

本田武司大阪大学, 微生物病研究所, 教授 (60029808)