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Purification and characterization of cytotoxin produced by H.phlori, and mechanisms for the occurrence of gastroduodenal diseases by H.pylori infection.

Research Project

Project/Area Number 05807022
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Bacteriology (including Mycology)
Research InstitutionKyorin University (1994)
Tokai University (1993)

Principal Investigator

KAMIYA Shigeru  Kyorin University School of Medicine, Department of Microbiology Professor, 医学部, 教授 (10177587)

Co-Investigator(Kenkyū-buntansha) YAMAGUCHI Hiroyuki  Kyorin University School of Medicine, Department of Microbiology, Research Assoc, 医学部, 助手 (40221650)
SHIRAI Takayuki  Tokai University School of Medicine, 6th Department of Internal Medicine, Resear, 医学部, 助手 (70196591)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1993: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsHelicobacter pylori / cytotoxin / chronic gastritis / peptic ulcer / isolation culture / urease / adherence / フローサイトメーター / 赤血球凝集活性 / 細胞傷害性毒素
Research Abstract

To examine association of Helicobacter pylori infection with the occurrence of gastroduodenal diseases, isolation of H.phlori from patients with the diseases was carried out. Isolation rates of H.pylori from the patients with gastric ulcer, duodenal ulcer and atrophic gastritis were 83,85 and 75%, respectively. In contrast, those from the patients with acute superficial gastritis and submucosal tumour were 25 and 33%, respectively. Sensitivity, specificity, positive^- and negative^-predictive values of rapid urease test of biopsy specimens to cultivation method were 76,85,91 and 63%, respectively, indicating that the rapid urease test is an useful screening test. By quantitative culture, it was shown that 5,000 - 10,000 CFU of H.phlori were required for positive reaction in the urease test.
Nearly half strains of H.phlori produce cytotoxin (CT) which induces vacuolation in cultured cells. It was demonstrated that CT was heat-labile at 70゚C and proteinous toxin with molecular weight of more than 20kDa. After precipitation of CT with 50% ammonium sulphate, purification of CT was tried using Sephacryl S300, phenylsuperose, Q Sepharose FF and Mono Q columns. However, highly purified CT was not obtained by these procedures. Positive rates for detection of CT using RK-13 (rabbit kidney), FL (human amnion), Vero (monkey kidney), BHK-21 (hamster kidney) and HeLa (human uterine cancer) cells were 73,61,27,27 and 21%, respectively, indicating that RK-13 cell line is a good indicator.
Adherence activity of H.phlori strains to human gastric cancer (MKN45, KATO III) cells and human intestine (Int-407) cells was analyzed by flowcytometer. Average adherence rates of H.pylori straius to MKN 45, KATO III and Int-407 cells were 76.6,42.7 and 15.1%, respectively. These results suggest that H.pylori has an affinity to gastric cells, particularly MKN 45 cells, rather than instinal cells. No significant correlation between adherence activity of and CT production by H.pylori was observed.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Kamiya S et al: "Evaluation of rapid urease test for detection of Helicobacter pylori in gastric biopsy specimens." Eur J Epidemiol. 9. 450-452 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 白井孝之 他: "Helicobacter pylori 陽性例の経過と治療." Current Therapy. 11. 99-104 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kamiya S et al: "Characteristics of vacuolating toxin produced by Helicobacter pylori." Eur J Gastroenterol Hepatol. 6. 923-27 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 高木敦司 他: "PCR法を用いたHelicobacter pylori感染診断に関する臨床的検討." 消化器病誌. 91. 1277-1282 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 神谷茂: "Helicobacter pyloriの生物学的特性." 医学のあゆみ. 169. 255-258 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yamamoto-Osaki T et al: "Adherence of Helicobacter pylori to cultured human gastric carcinoma cells." Eur J Gastroenterol Hepatol. (in press). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 白井孝之 他: "Helicobacter pylori(Hp)の存在診断におけるbiopsy urease test (BUT)の有用性“Helicobacter pyloriによる胃十二指腸粘膜障害の臨床的意義"" トプコ出版部, 127 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kamiya S.et al.: "Evaluation of rapid urease test for detection of Helicobacter pylori in gastric biopsy specimens" Eur J Epidemiol. 9. 450-452 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shirai T.et al.: "The role of Helicobacter pylori in pathophysiology of gastro-duodenal ulcer (in Japanese)" Rinsho-shokaki-naika. 8. 541-547 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shirai t.et al.: "Clinical courses and therapy for the patients infected with Helicobacter pylori (in Japanese)" Current Therapy. 11. 99-104 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kamiya S.et al.: "Characteristics of vacuolating toxin produced by Helicobacter pylori" Eur J Gastroenterol Hepatol. 6 (Suppl 1). 23-27 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Takagi A.et al.: "Evaluation of polymerase chain reaction for diagnosis of Helicobacter pylori infection (in Japanese)" Shokakibyo-zasshi. 91. 1277-1282 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kobayashi H.et al.: "Effect of various proton pump inhibitors on growth of Helicobacter pylori ; comparison to productivity of vacuolating toxin (in Japanese)" Rinsho to Biseibutsu. 21. 607-611 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kamiya S.et al.: "Biological characteristics of Helicobacter pylori (in Japanese)" Igaku no ayumi. 169. 255-258 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shirai T.et al.: "Helicobacter pylori and gastroduodenal ulcer (in Japanese)" Shindan to chiryou. 82. 1281-1284 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yamamoto-Osaki et al.: "Adherence of Helicobacter pylori to cultured human gastric carcinoma cells" Eur J Gastroenterol Hepatol. (in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shirai T.et al.: "Usefulness of biopsy urease test (BUT) for diagnosis of the presence of Helicobacter pylori (in Japanese)" In : Clinical significance of gastroduodenal mucosal injury by Helicobacter pylori infection, (ed.by Miwa T.) Topuko Publishing (in Japanese). 22-29 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kamiya S,et al: "Evaluation of rapid urese test for detection of Helicobacter pylori in gastric biopsy specimens." Eur J Epidemiol. 9. 450-452 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] Kamiya S,et al: "Characteristics of vacuolating toxin produced by Helicobacter pylori." Eur J Gastroenterol Hepatol. 6. S23-27 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yamamoto-Osaki T,et al: "Adherence of Helicobacter pylori to cultured human gastric carcinoma cells." Eur J Gastroenterol Hepatol. (in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 神谷 茂: "Helicobacter pyloriの生物学的特性." 医学のあゆみ. 169. 255-258 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 白井孝之,他: "胃潰瘍および十二指腸潰瘍におけるHPの位置づけ." 臨床消化器内科. 8. 541-547 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] 高木敦司,他: "PCR法を用いたHelicobacter pylori感染診断に関する臨床的検討." 消化器病誌. 91. 1277-1282 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 白井孝之,他: "Helicobacter pylori(HP)の存在診断におけるbiopsy ureasetest(BUT)の有用性“Helicabacter pylorによる胃十二指腸粘膜障害の臨床的意義"" トプコ出版部, 127 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] S.Kamiya: "Evaluation of rapid urease test for detection of Helicobacter pylori in gastric biopsy specimens" European Journal of Epidemiology. 9. 450-452 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] S.Kamiya: "Characteristics of vacuolating toxin produced by Helicobacter pylori" European Journal of Gastroenterology and Hepatology. (in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 神谷茂: "Helicobacter pyloriの生物学的特性" 医学のあゆみ. (印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 高木敦司: "PCR法を用いたHelicobacter pylori感染診断に関する臨床的検討" 日本消化器病学会誌. (印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] S.Kamiya: "Analysis of intestinal flora of a patient with congenital absence of the portal vein" FEMS Immunology and Medical Microbiology. 7. 73-80 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 白井孝之: "Helicobacter pyloriによる胃十二指腸粘膜障害の臨床的意義" トプコ印刷, 128 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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