DETERMINANTS OF B BURGDORFERI HEMATOGENOUS DISSEMINATION

描述(改编自申请者摘要)：最常见的莱姆病 在美国流行的节肢动物传播疾病，是由感染引起的 有螺旋体伯氏疏螺旋体。尽管最近进行了密集的研究 多年来，人们对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，伯氏杆菌 从早期莱姆病患者身上获得的Sensu Strigto分离株可以是 根据几个基因类型参数来区分，并且 这些基因型在培养的菌株中的分布有显著差异。 从早期莱姆病患者的皮肤或血液中提取。这表明 不同的伯氏杆菌基因型别在致病潜力上存在差异 血流传播。研究人员假设， 血源性传播的可能性是基因变异的结果 在不同基因型别中的表达，并建议确定其影响因素 这些差异的基础是基因组和蛋白质组的结合 接近了。通过这种分析确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) 用HIGH方法评估不同基因类型之间的基因表达差异 伯氏杆菌细胞总蛋白的分辨率分析。那些蛋白质 这些基因是特定基因类型所特有的，或者在 表达(在特定生长条件下)将通过以下方式标识 伯氏杆菌基因组的微测序及序列比较 数据库；2)通过HIGH分析选定的一组基因的表达 密度膜阵列技术及RT-PCR法检测 生理条件的变化；3)分析 直接在皮肤中进行RT-PCR分析鉴定出可能的毒力决定因素 莱姆病患者的活检组织和血液以及莱姆病患者的组织 实验感染的小鼠。功能基因组学与基因工程的结合 建议的VIVO方法应提供对 芽孢杆菌不同基因变种致病的分子基础 伯格多费氏菌具有严格的意义并应导致决定因素的鉴定 螺旋体传播所需的。这应该为我们提供新的见解 伯氏杆菌在人类中感染的自然历史，进而可能 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.