Vascular tropism of Borrelia burgdorferi

伯氏疏螺旋体的血管趋向性

• 批准号: 6871971
• 负责人: Jenifer L Coburn
• 金额: $ 28.35万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6871971
• 关键词: Borrelia; Lyme disease; SDS polyacrylamide gel electrophoresis; bacteria infection mechanism; bacterial DNA; bacterial proteins; enzyme linked immunosorbent assay; heart; host organism interaction; immunofluorescence technique; joints; laboratory mouse; peptide library; phage display; polymerase chain reaction; recombinant proteins; tissue /cell culture; tissue tropism; vascular endothelium; virulence; western blottings

DESCRIPTION (provided by applicant): This application for an NIH R21 award under PA-03-080 (Biodefense and Emerging Infectious Disease Research Opportunities) is based on preliminary studies just recently obtained in the laboratory, and that we believe are of great interest as the basis for further exploratory studies. Our laboratory is involved in the identification of virulence determinants of Borrelia burgdorferi, the causative agent of Lyme disease. Lyme disease is considered an emerging infection because the geographic distribution and the number of cases continue to expand in North America. The ability of B. burgdorferi to disseminate from the site of inoculation, a tick bite, to multiple tissues indicates that interactions with mammalian cells occur at multiple stages of infection. In particular, the hematogenous dissemination of the spirochete to multiple tissues suggests that interactions with the vascular endothelium are critical to the ability of B. burgdorferi to cause infection. As described in this proposal, we used a filamentous phage display library of B. burgdorferi DNA in vivo to select for phage clones that exhibit tropism to the vasculature in the hearts and joints of mice. These tissues were selected for analysis because they are frequently infected by B. burgdorferi in humans and in animal models of Lyme disease. We have identified three known or predicted outer membrane proteins of B. burgdorferi as potential vascular tropism determinants: OspC, VlsE, and the as yet uncharacterized product of gene BB0210. We now propose to generate the relevant fragments of these three proteins in recombinant form, and to test the recombinant proteins for specific binding to endothelial cells. In addition, we will investigate the nature of the endothelial cell receptors for these B. burgdorferi proteins. The experiments that we are proposing will shed light on the significance of three B. burgdorferi proteins that may facilitate interactions of B. burgdorferi with the vasculature during the natural course of infection in mammals.

描述（由申请人提供）：这份 PA-03-080（生物防御和新兴传染病研究机会）下的 NIH R21 奖项申请是基于最近在实验室获得的初步研究，我们认为这些研究非常有趣为进一步探索性研究奠定基础。我们的实验室致力于鉴定莱姆病的病原体伯氏疏螺旋体的毒力决定因素。莱姆病被认为是一种新出现的感染，因为北美地区的地理分布和病例数量持续扩大。伯氏疏螺旋体从接种部位（蜱叮咬处）传播到多个组织的能力表明，与哺乳动物细胞的相互作用发生在感染的多个阶段。特别是，螺旋体通过血行传播到多个组织表明，与血管内皮的相互作用对于伯氏疏螺旋体引起感染的能力至关重要。如本提案中所述，我们在体内使用了伯氏疏螺旋体 DNA 的丝状噬菌体展示文库来选择对小鼠心脏和关节的脉管系统表现出趋向性的噬菌体克隆。选择这些组织进行分析是因为它们在人类和莱姆病动物模型中经常被伯氏疏螺旋体感染。我们已经鉴定出三种已知或预测的伯氏疏螺旋体外膜蛋白作为潜在的血管向性决定因素：OspC、VlsE 和尚未表征的基因 BB0210 产物。我们现在建议以重组形式产生这三种蛋白的相关片段，并测试重组蛋白与内皮细胞的特异性结合。此外，我们将研究这些伯氏疏螺旋体蛋白的内皮细胞受体的性质。我们提出的实验将揭示三种伯氏疏螺旋体蛋白的重要性，这些蛋白可能促进哺乳动物自然感染过程中伯氏疏螺旋体与脉管系统的相互作用。