Traversal mechanism of epithelial cells by P. aeruginosa

铜绿假单胞菌穿越上皮细胞的机制

• 批准号: 7172219
• 负责人: IRANIA ALARCON
• 金额: $ 2.99万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7172219
• 关键词: Bacteria; Bacterial Pili; Biological Assay; Cells; Cessation of life; Contact Lenses; Cornea; Data; Depth; Development; Disease; Epithelial; Epithelial Cells; Fluorescein; Fluoresceins; Fluorescence Microscopy; Gentamicins; Goals; Green Fluorescent Proteins; Immunohistochemistry; Infection; Injury; Invaded; Label; Laboratories; Lead; Life; Measurement; Measures; Methods; Movement; Mus; Pathogenesis; Permeability; Pilum; Plaque Assay; Play; Positioning Attribute; Predisposing Factor; Property; Pseudomonas aeruginosa; Relative (related person); Role; Role playing therapy; Sampling; Speed; Standards of Weights and Measures; Surface; Testing; Thick; Tight Junctions; Time; Tracer; Travel; Video Microscopy; Virulence; Vision; cell motility; corneal epithelium; extracellular; in vivo; insight; monolayer; mutant; time use

DESCRIPTION (PROVIDED BY APPLICANT): Corneal infection caused by P. aeruginosa (PA) is a sight threatening disease that is difficult to treat. Contact lens wear, the most common predisposing factor for this infection, does not normally cause full thickness epithelial injury to the cornea. Yet little is known about mechanisms that PA use to traverse corneal epithelial cells and reach the underlying stroma. Since traversal represents an early step in pathogenesis, insight into the mechanisms involved could lead to development of preventative measures that completely avoid the devastating effects of PA infection. Twitching motility (tm), a property of bacterial pili, has been found to play a role in corneal infection caused by PA; preliminary data show it is required for traversal across corneal epithelial cells in culture. This project will be focused on the mechanisms by which tm contributes to traversal. The hypothesis to be tested is that tm plays a role in the exit of PA from cells it has invaded and that it is also involved in travel between cells. Exit from corneal epithelial cells will be studied using standard plaque assays, modified gentamicin survival assay and quantitative deconvolution fluorescence microscopy (dm). Extracellular travel will be studied using time-lapse video microscopy and dm.

描述（由申请人提供）：由铜绿假单胞菌（PA）引起的角膜感染是一种难以治疗的视力威胁性疾病。接触透镜配戴是这种感染最常见的诱发因素，通常不会对角膜造成全层上皮损伤。然而，PA用于穿过角膜上皮细胞并到达底层基质的机制知之甚少。由于穿越代表了发病机制的早期步骤，因此对所涉及机制的深入了解可能会导致预防措施的发展，从而完全避免PA感染的破坏性影响。抽搐运动性（Twitching motility，tm）是细菌皮利的一种特性，已被发现在PA引起的角膜感染中起作用;初步数据显示，它是穿过培养物中的角膜上皮细胞所必需的。这个项目将集中在机制，通过该机制TM有助于遍历。待检验的假设是，TM在PA从其侵入的细胞中退出中起作用，并且它还参与细胞之间的旅行。将使用标准菌斑试验、改良庆大霉素存活试验和定量去卷积荧光显微镜（dm）研究角膜上皮细胞的退出。细胞外旅行将使用延时视频显微镜和dm研究。