POLYMORPHIC MEMBRANE PROTEINS OF CHLAMYDIA TRACHOMATIS

描述(申请人提供)：衣原体多形膜蛋白(PMPs)是一个新发现的衣原体特异性膜蛋白家族，其在衣原体生物学和发病机制中的作用尚不清楚。对肺炎衣原体PMP家族的基因组分析发现了移码突变、缺失和基因复制。对肺炎链球菌和鹦鹉热杆菌的PMP大家族的研究也表明，PMP蛋白在体外表达，一些可以在基本体表检测到，一些在感染过程中是优势抗原，可能成为疫苗设计的靶标。新出现的证据与PMP家族在发病机制和免疫逃避中的作用是一致的。 这个项目的目的是描述迄今为止发现的最小的PMP家族：沙眼衣原体PMP家族的9个成员。在使用这9个部分纯化的重组PMP作为靶抗原的初步研究中，我们观察到盆腔炎患者存档血清中PMP特异性抗体反应的差异。这一分析将通过横断面和纵向比较生殖器沙眼衣原体L感染患者群体中PMP特异性反应来扩展。这一分析可能确定PMP特异性反应和疾病结局之间的直接关系。更重要的是，这一分析将为进一步的分子鉴定提供一套新鲜的临床沙眼衣原体分离株。该项目的第二个重点将是确定和表征沙眼衣原体中PMP表达的决定因素。将在选定的研究菌株的PMP家族中识别和比较多态。将进行实验，以表征这些分离物中PMP表达的发育模式。利用本项目中产生的一组PMP特异性的单抗和多克隆抗体，我们将利用激光扫描共聚焦荧光显微镜在单细胞水平上检测PMP蛋白的表达，并最终在单细胞水平上将PMP蛋白转移到外膜表面。

DESCRIPTION (provided by applicant): Chlamydial polymorphic membrane proteins (Pmps) are a newly identified family of Chlamydia-specific membrane proteins, whose role in chlamydial biology and pathogenesis is unknown. Genomic analysis of the pmp family of C. pneumoniae have revealed frameshift mutations, deletions and gene duplications. Studies of the larger pmp families of C. pneumoniae and C. psittaci have also revealed that Pmp proteins are expressed in vitro, that some can be detected at the elementary body surface, and that some are dominant antigens during infection and may be targets for vaccine design. The emerging evidence is consistent with a role of the pmp family in pathogenesis and immune evasion. The purpose of this project is to characterize the smallest pmp family identified to date: the 9-member pmp family of C. trachomatis. In preliminary studies using the 9 partially purified recombinant Pmps as target antigens, we have observed differential Pmp-specific antibody responses in archived sera from patients with pelvic inflammatory disease. This analysis will be expanded through cross-sectional and longitudinal comparisons of Pmp-specific responses in a well-characterized patient population with genital C. trachomatis l infection. This analysis may identify direct relationships between Pmp-specific responses and disease outcome. More importantly, this analysis will provide a set of fresh clinical C. trachomatis isolates for further I molecular characterization. A second focus of this project will be to identify and characterize determinants of pmp expression in C. trachomatis. Polymorphisms will be identified and compared in the pmp families of selected study isolates. Experiments will be performed to characterize developmental patterns of pmp expression in these isolates. Using a panel of Pmp-specific monoclonal and polyclonal antibodies generated in this project, we will examine Pmp protein expression and eventual translocation to the surface of the outer membrane along development and at the single cell level using laser scanning confocal fluorescence microscopy.