ANALYSIS OF INTERACTION BETWEEN HELICOBACTER AND GASTRC EPITHELIAL CELLS BY YEAST TWO-HYBRID METHOD

酵母双杂交法分析螺杆菌与胃上皮细胞的相互作用

• 批准号: 12670449
• 负责人: SUGIYAMA Toshiro
• 金额: $ 2.5万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670449/
• 关键词: HELICOBACTER PYLOR; OUTER MEMBRANE; ADHESION; TYPE 4 SECRETION MACHINERY; アポトーシス

The aim of this study is to investigate the molecular mechanism of interactions between Helicobacter pylori(H.pylori) infection and gastric epithelial cells by using yeast two-hybrid method. Consequently, 45 genes of H.pylori associated with an adhesion to gastric epithelial cells and cagG gene within cag pathogenicity(cagPAI) were selected. In Japanese strains, only 4 % of the tested clinical isolates had partially deleted cagPAI genes of H.pylori. All of them had deleted cagG of cagPAI. Although cagPAI functions as type 4 secretion machinery to insert some proteins or DNAs of H.pylori into an epithelial cell, cagGgene is not speculated to be included in the component of type 4 secretion machinery. However, all cagG deleted strains demonstrated low induction of IL-8 from gastric epithelial cells and low induction of NF-kB. As cagG gene has a homology with flagella motor switch protein gene of the other bacteria, we tested an adhesion ability to gastric epithelial cells by flow-cytometory. All cagG deleted strains showed low adhesion ability to the gastric epithelial cells. As we were able to construct cagG knock- out mutant, these results should be confirmed by using isogenic mutant.

本研究旨在利用酵母双杂交技术探讨幽门螺杆菌（H.pylori）感染与胃上皮细胞相互作用的分子机制。因此，选择了45个与胃上皮细胞粘附相关的幽门螺杆菌基因和cag致病性内的cagG基因（cagPAI）。在日本菌株中，只有4%的测试临床分离株具有部分缺失的幽门螺杆菌cagPAI基因。所有患者cagPAI的cagG基因均缺失。尽管cagPAI作为4型分泌机制将幽门螺杆菌的一些蛋白质或DNA插入上皮细胞，但推测cagG基因不包括在4型分泌机制的组分中。然而，所有cagG缺失的菌株表现出从胃上皮细胞低诱导IL-8和低诱导NF-κ B。由于cagG基因与其他细菌的鞭毛马达开关蛋白基因具有同源性，我们通过流式细胞术检测了对胃上皮细胞的粘附能力。所有cagG缺失菌株对胃上皮细胞的粘附能力均较低。由于我们能够构建cagG基因敲除突变体，这些结果需要通过使用同基因突变体来证实。

项目成果

Mizushima T, Sugiyama T. et al.: "Clinical relevance of the babA2 genotype of Helicobacter pylori in Japanese clinical isolates"J Clin Microbil. 39. 2463-2465 (2001)

Mizushima T、Sugiyama T. 等人：“日本临床分离株中幽门螺杆菌 babA2 基因型的临床相关性”J Clin Microbil。

Sugiyama T. et al.: "Attributable risk of H.pylori in peptic ulcer diseases : Does the declining prevalence of infection in general population explain the increasing frequency of non-H.pylori ulcer?"Dig Dis Sci. 46. 307-310 (2001)

Sugiyama T. 等人：“消化性溃疡疾病中幽门螺杆菌的归因风险：普通人群中感染率的下降是否可以解释非幽门螺杆菌溃疡发生率的增加？”Dig Dis Sci。

Hayashi S, Sugiyama T.: "Combined effect of rebamipide and…"Microbiol Immunol. 44. 557 (2000)

Hayashi S、Sugiyama T.：“瑞巴派特和……的综合作用”微生物免疫学。 44. 557 (2000)

Kato S, Sugiyama T.: "CagA antibody in Japanese children…"J.Clin Microbiol. 38. 68 (2000)

Kato S, Sugiyama T.：“日本儿童中的 CagA 抗体……”J.Clin Microbiol。38. 68 (2000)

Sugiyama T, Nishikawa K, Asaka M, Freston J.: "Attributable risk of H.pylori in peptic ulcer diseases :Does the declining prevalence of infection in general population explain the increasing frequency of non-H.pylori ulcer?"Dig Dis Sci. 46. 307-310 (2001)

Sugiyama T、Nishikawa K、Asaka M、Freston J.：“消化性溃疡疾病中幽门螺杆菌的归因风险：普通人群中感染率的下降是否可以解释非幽门螺杆菌溃疡发生率的增加？”Dig Dis Sci