BACTERIAL INVASION OF EUKARYOTIC TISSUES

真核组织的细菌入侵

• 批准号: 5200696
• 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/5200696
• 关键词: bacterial cytopathogenic effect; bacterial genetics; confocal scanning microscopy; enteric bacteria; eukaryote; fluorescence microscopy; 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都是早期和 疾病发生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年代初的工作确立了S 志贺氏菌触发入侵的遗传机制编码在一个 毒力相关质粒。最近的研究表明， 其他肠道细菌的入侵能力是由染色体编码的。 本项目旨在了解原核生物和真核生物 对细菌内化的要求，最终目标是 对细菌所涉及的事件的彻底分子定义 真核组织的入侵。目前的实验方法包括 细菌进入人体培养细胞系的检测方法的应用 不同组织来源的上皮细胞。生物化学抑制物 原核细胞的结构/功能或真核细胞过程 在这些组织培养侵袭试验中使用来检查 对细菌摄取的要求。细菌的直接可视化 通过透射式电子显微镜测量进入，视频 显微镜、荧光显微镜和共聚焦显微镜。 除了上面描述的机械方法，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定的真核细胞 通过抑制物竞争来测量对细菌入侵的反应 感染真核生物的检测、配基结合分析和mRNA分析 细胞。从这些方法中的每一种获得的信息都被集成 为每个人提供对细菌进入的机械理解 研究了这种途径。