Infection mechanism of E.coli 0157 to native human colon epithelial cells

大肠杆菌0157对天然人结肠上皮细胞的感染机制

• 批准号: 12670268
• 负责人: YOSHIDA Tomoaki
• 金额: $ 2.05万
• 依托单位: Aichi Medical 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-12670268/
• 关键词: Enterohemorrhagic E. coli; Colon epithelial cells; Intracelluar invasion; Type 3 secretion; Cytoskeleton; 細胞内感染; インティミン; type III分泌; typeIII分泌

The purpose of this project is to elucidate the infection mechanism of enterohemorrhagic E. coli O157 to human intestinal epithelial cells.1) The primary culture of native epithelial cells from human colon has been established.2) E. coli O157 has been shown to invade the native colon epithelial cells to a comparable extent with Salmonella enterica as proved by killing the extracellular bacteria by gentamicin. The invasion was dependent on the action of cytoskeletons and cell surface cholesterol content.3) The morphological insights using con focal fluorescent microscopy and electromicroscopy confirmed the intracellular bacteria, in addition to gentamicin protection assay.4) The target cells of E. coli O157 were shown to express epithelial markers but not monocyte markers.5) When another type of epithelial cell, Hela was tested for the infection of E. coli 0157 and salmonella, the latter invaded to a similar degree of colon cells but not the former.6) Employing the isogenic deletion mutants of bacterial effacement locus, the sepL was proved to be essential for the invasion activity, whereas not was eaeA, espA or Tir.

本课题旨在阐明肠出血性大肠杆菌的感染机制。coli O157对人肠上皮细胞的体外培养，建立了原代培养人结肠上皮细胞的方法。大肠杆菌O157已经显示出侵入天然结肠上皮细胞的程度与肠道沙门氏菌相当，这通过庆大霉素杀死细胞外细菌来证明。3）共聚焦荧光显微镜和电子显微镜观察证实了胞内细菌的存在，庆大霉素保护试验也证实了E. coliO 157表达上皮细胞标记而不表达单核细胞标记。coli 0157和SaleA对大肠杆菌的侵袭能力相似，而SaleA对大肠杆菌的侵袭能力较低。6）利用细菌缺失位点的等基因缺失突变体，证实了sepL是大肠杆菌侵袭能力所必需的，而eaeA、espA和Tir则不是必需的。

项目成果

D.Chakravortty, Y.Kato, T.Sugiyama, N.Koide, M.M.Mu, T.Yoshida, T.Yokochi.: "Inhibition of p38 mitogen-activated protein kinase augments lipopolysaccharide-induced cell proliferation in CD14-expressing Chinese hamster ovary cells"Infect.Immun. 69. 931-936

D.Chakravortty、Y.Kato、T.Sugiyama、N.Koide、M.M.Mu、T.Yoshida、T.Yokochi.：“抑制 p38 丝裂原激活蛋白激酶可增强表达 CD14 的中国仓鼠卵巢中脂多糖诱导的细胞增殖

T.Yoshida, T.Chiba, T.Yokochi, K Onozaki, T.Sugiyama, I.Nakashima.: "Synthesis of a set of di-and tri-sulfated galabioses"Carbohydr Res.. 335. 167-180 (2001)

T.Yoshida、T.Chiba、T.Yokochi、K Onozaki、T.Sugiyama、I.Nakashima.：“一组二-和三-硫酸化半乳糖的合成”碳水化合物研究.. 335. 167-180 (2001)

T.Yoshida, T.Chiba, T.Yokochi, K.Onozaki, T.Sugiyama, I.Nakashima.: "Synthesis of a set of di-and tri-sulfated galabioses"Carbohydr Res.. 335. 167-180 (2001)

T.Yoshida、T.Chiba、T.Yokochi、K.Onozaki、T.Sugiyama、I.Nakashima.：“一组二硫酸化和三硫酸化半乳糖的合成”碳水化合物研究 335. 167-180 (2001

D.Chakravortty, Y.Kato, T.Sugiyama, N.Koide, M.M.Mu, T.Yoshida, T.Yokochi.: "The inhibitory action of sodium arsenite on lipopolysaccharide-induced nitric oxide production in RAW 267.4 macrophage cells: A role of Raf-1 in LPS signaling"J.Immunol.. 166. 20

D.Chakravortty、Y.Kato、T.Sugiyama、N.Koide、M.M.Mu、T.Yoshida、T.Yokochi.：“亚砷酸钠对 RAW 267.4 巨噬细胞中脂多糖诱导的一氧化氮产生的抑制作用：一个作用

Chakravortty D, Kato Y, Sugiyama I, Koide N, Mu MM, Yoshida T, Yokochi T.: "Inhibition of Caspase 3 Abrogates Lipopolysaccharide-Induced Nitric Oxide Production by Preventing Activation of NF-kappaB and c-Jun NH(2)-Terminal Kinase/Stress-Activated Protein

Chakravortty D、Kato Y、Sugiyama I、Koide N、Mu MM、Yoshida T、Yokochi T.：“抑制 Caspase 3 通过阻止 NF-kappaB 和 c-Jun NH(2)- 的激活来消除脂多糖诱导的一氧化氮产生