Polymorphic membrane proteins of Chlamydia trachomatis

描述（由申请人提供）：衣原体多态膜蛋白（pmp）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。肺炎C. prop家族的基因组分析揭示了移码突变、缺失和基因重复。对肺炎C.和鹦鹉热C.较大的pmp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，一些是感染期间的显性抗原，可能是疫苗设计的靶点。新出现的证据与prop基因家族在发病机制和免疫逃避中的作用一致。这个项目的目的是表征迄今为止发现的最小的pmp基因家族：沙眼衣原体的9个成员家族。在使用9种部分纯化的重组Pmps作为靶抗原的初步研究中，我在盆腔炎患者的存档血清中观察到不同的pmp特异性抗体反应。该分析将通过对生殖器沙眼衣原体感染患者群体中pmp特异性反应的横断面和纵向比较进行扩展。该分析可以确定pmp特异性反应与疾病结局之间的直接关系。该项目的第二个重点将是确定和表征沙眼衣原体中Pmp表达的遗传和分子决定因素。将在选定的研究分离株中鉴定和比较pmp家族的多态性。实验将进行表征发育模式的pmp表达在这些分离。使用本项目中生成的Pmp特异性单克隆和多克隆抗体，我将使用激光扫描共聚焦荧光显微镜在单细胞水平上检查Pmp蛋白的表达和最终外膜表面的易位。

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 prop family of C. pneumoniae has 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 prop gene family in pathogenesis and immune evasion. The purpose of this project is to characterize the smallest pmp gene family identified to date: the 9-member family of C. trachomatis. In preliminary studies using the 9 partially purified recombinant Pmps as target antigens, I 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 infection. This analysis may identify direct relationships between Pmp-specific responses and disease outcome. A second focus of this project will be to identify and characterize genetic and molecular 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, I 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.