BACTERIAL INVASION OF EUKARYOTIC TISSUES

真核组织的细菌入侵

• 批准号: 6161214
• 负责人: D J KOPECKO
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6161214
• 关键词: bacterial cytopathogenic effect; bacterial genetics; confocal scanning microscopy; enteric bacteria; eukaryote; fluorescence microscopy; gastrointestinal epithelium; gastrointestinal infection; messenger RNA; molecular cloning; molecular pathology; pathologic process; prokaryote; receptor binding; tissue /cell culture; transmission electron microscopy; video microscopy; virulence

Many bacterial pathogens of the intestinal tract have as an early and necessary step in initiation of disease the ability to penetrate gut epithelial cells. My lab's work in the early 1980's established that Shigella's genetic machinery to trigger invasion is encoded on a large virulence-associated plasmid. More recent studies have shown that the invasion ability of other enteric bacteria is chromosomally encoded. This project is aimed at understanding the prokaryotic and eukaryotic requirements for bacterial internalization, with the ultimate aim being a thorough molecular definition of the events involved in bacterial invasion of eukaryotic tissues. Current experimental approaches involve the use of assays measuring bacterial entry into cultured lines of human epithelial cells of various tissue origins. Biochemical inhibitors of prokaryotic structure/function or of eukaryotic cell processes are employed in these tissue culture invasion assays to examine the requirements for bacterial uptake. Direct visualization of bacterial entry is measured via transmission electron microscopy,video microscopy,fluorescent microscopy, and confocal microscopy. In addition to the mechanistic approaches described above, genetic techniques are employed to clone the responsible bacterial genes. Eukaryotic receptors for bacterial ligands and specific eukaryotic cell responses to bacterial invasion are measured via inhibitor competition assays, ligand binding assays, and mRNA analyses of infected eukaryotic cells. The information gained from each of these approaches is integrated to provide a mechanistic understanding of bacterial entry for each pathway studied.

肠道的许多细菌病原体具有早期和早期的特征， 在疾病开始的必要步骤，穿透肠道的能力， 上皮细胞我的实验室在20世纪80年代早期的工作表明， 志贺氏菌触发入侵的遗传机制编码在一个大的 毒力相关质粒。最近的研究表明， 其它肠道细菌的侵入能力是染色体编码的。 本项目旨在了解原核和真核生物 细菌内化的要求，最终目的是 对细菌入侵所涉及的事件进行彻底的分子定义 真核生物组织。目前的实验方法涉及使用 测量细菌进入人上皮细胞培养系的测定 各种组织来源的细胞。原核生物的生化抑制剂 结构/功能或真核细胞过程 组织培养侵入试验，以检查细菌的要求 摄取。细菌进入的直接可视化是通过 透射电子显微镜，视频显微镜，荧光显微镜， 和共聚焦显微镜。 除了机械的方法， 如上所述，遗传技术被用来克隆负责的 细菌基因细菌配体的真核受体和特异性 通过抑制剂测定真核细胞对细菌入侵的反应 竞争测定、配体结合测定和感染的 真核细胞从这些方法中获得的信息是 整合，以提供细菌进入的机制理解， 每一个研究的路径。