DETERMINANTS OF B BURGDORFERI HEMATOGENOUS DISSEMINATION

描述（改编自申请人的摘要）：莱姆病，最常见的疾病 在美国流行的节肢动物传播的疾病，是由感染引起的 螺旋体伯氏疏螺旋体尽管近年来进行了深入研究， 多年来，关于感染的分子发病机制知之甚少， 水平在最近的研究中，研究人员表明，B。burgdorferi 从早期莱姆病患者获得的严格意义上的分离物可以 根据几个基因型参数进行区分， 这些基因型的分布在培养的分离物中显著不同 从莱姆病早期患者的皮肤或血液中提取。这表明 不同基因型的B。严格意义上的burgdorferi sensu stricto在它们的潜力上各不相同， 血流播散研究人员假设， 血源性传播的可能性是基因变异的结果 不同基因型之间的表达，并提出确定因素 通过基因组学和蛋白质组学的结合， 接近。通过这种分析鉴定的基因的表达将被 在莱姆病患者中进行监测。该项目的具体目标是：1） 评估不同基因型之间的基因表达差异， 总B的分辨率分析。burgdorferi细胞蛋白那些蛋白质 其对于特定基因型是独特的，或者其在以下方面显示出显著的变化： 表达（在特定生长条件下）将通过以下方法鉴定： 微测序并与B中的序列进行比较。伯氏基因组 数据库; 2）通过高表达分析所选基因组的表达， 密度膜阵列技术和RT-PCR的分离培养下， 3）分析了不同生理条件下， 通过直接在皮肤中进行RT-PCR分析确定的推定毒力决定因素 莱姆病患者的活检组织和血液以及 实验感染的老鼠功能基因组学和基因组学的结合 所提议的体内方法应规定全面评估 B不同基因型变异发病机制的分子基础。 严格意义上的burgdorferi，并应导致确定决定因素 是螺旋体传播所必需的这将为我们提供新的见解， B的自然史人类感染伯氏螺旋体，反过来， 对莱姆病的预防、治疗和诊断具有重要意义。

DESCRIPTION (Adapted from the Applicant's Abstract): Lyme disease, the most prevalent arthropod-borne disease in the United States, is caused by infection with the spirochete, Borrelia burgdorferi. Despite intensive study in recent years little is known regarding the pathogenesis of infection at the molecular level. In recent studies, the investigators have shown that B. burgdorferi sensu stricto isolates obtained from patients with early Lyme disease can be distinguished on the basis of several genotypic parameters and that the distribution of these genotypes is significantly different in isolates cultured from the skin or blood of patients with early Lyme disease. This suggests that different genotypes of B. burgdorferi sensu stricto vary in their potential for bloodstream dissemination. The investigators hypothesize that differences in potential for hematogenous spread are the result of variations in gene expression among different genotypes and propose to identify the factors underlying these differences by a combination of genomic and proteomic approaches. The expression of genes identified by such analysis will then be monitored in Lyme disease patients. The specific aims of the project are: 1) to assess differences in gene expression among the various genotypes by high resolution analysis of total B. burgdorferi cellular protein. Those proteins which are unique to specific genotypes or which show significant variations in expression (under specific growth conditions) will be identified by microsequencing and comparison to sequences in the B. burgdorferi genome database; 2) to analyze expression for a selected group of genes by high density membrane arrays techniques and by RT-PCR of isolates cultured under a variety of physiological conditions; 3) to analyze the expression of identified, putative virulence determinants by RT-PCR analysis directly in skin biopsy tissue and blood of Lyme disease patients and in tissues of experimentally infected mice. The combination of functional genomics and in vivo approaches proposed should provide for the comprehensive assessment of the molecular basis of pathogenesis for different genotypic variants of B. burgdorferi sensu stricto and should result in identification of determinants required for spirochete dissemination. This should provide new insights into the natural history of B. burgdorferi infection in humans, which, in turn, may have implications for prevention, treatment, and diagnosis of Lyme disease.