STUDIES OF CHLAMYDIA TRACHOMATIS

沙眼衣原体是细菌获得性行为的主要原因 在美国传播感染，并对25%-50%的 据估计，每年有100万例盆腔炎。 后遗症包括宫外孕和不孕症。为了开始剖析 破坏性与保护性反应的对比，以及识别潜力 候选疫苗，我们的总体目标是了解细胞和 感染衣原体抗原分选和分泌的分子生物学研究 极化的人子宫内膜上皮细胞。 我们检测到衣原体内毒素和主要外膜的逃逸 蛋白质(MOMP)在感染细胞表面之前自然 在发育周期结束时衣原体的释放。做 衣原体应激反应蛋白也会逃逸到受感染的细胞 表面：这些抗原是通过什么途径从包涵体中获得的？ 表面暴露的抗原是由宿主免疫反应感知的吗： 我们将使用强大的包埋后免疫电子技术 Lowicryl加工样品的显微镜，以确定特定的目标1 沙眼衣原体血清型E、GroEL和DNAK也分泌到表面 正常感染和青霉素和干扰素诱导的 持续感染人子宫内膜上皮细胞，在特定情况下 目的2继续评估衣原体抗原是否为 以两极分化的方向分泌。一抗(ABY)定向 针对衣原体，特异组蛋白将作为控制标志 不能逃脱包涵体的抗原和Maby针对的 1类分子将作为基侧结构域的标记。 复制样本的数字化荧光显微镜将提供更多 对定向抗原分泌模式的定量评估。在……里面 具体目标3，我们将调查衣原体表面抗原 分泌受雌激素和黄体酮的调节。因为跨境- 高尔基体胞吐抑制剂Brefeldin A减少内毒素和MOMP的逃逸 包涵体、衣原体抗原可能利用分泌途径到达 被感染的细胞表面。在具体目标4中，我们将分析 共定位逃逸衣原体对衣原体抗原分选途径的研究 抗原与高尔基体标记物β-COP、p58和C6-NBD-神经酰胺。 最后，为了开始研究衣原体生长的免疫控制，我们有 建立体外共培养模型评价宿主效应PMN 以及T细胞对受感染的人类上皮细胞的反应。

C. trachomatis is the leading cause of bacerially-acquired sexually transmitted infections in the USA and is responsible for 25-50% of the estimate 1 million cases per year of pelvic inflammatory disease. Sequelae include ectopic pregnancy and sterility. To begin to dissect the destructive versus the protective response as well as identify potential vaccine candidates, our overall goal is to understand the cell and molecular biology of chlamydial antigen sorting and secretion in infected polarized human endometrial epithelial cells. We have detected escape of chlamydial LPS and the major outer membrane protein (MOMP) to the surface of infected cells prior to the natural release of chlamydiae at the end of the developmental cycle. Do chlamydial stress response proteins also escape to the infected cell surface: What pathway do these antigens take from the inclusion and how are the surface exposed antigens perceived by the host immune response: We shall use the powerful technique of post-embedding immunoelectron microscopy on Lowicryl-processed samples to determine in Specific Aim 1 if C. trachomatis serovar E GroEL and DnaK are also secreted to the surface of normally infected and penicillin and interferon gamma-induced persistently infected human endometrial epithelial cells, and in Specific Aim 2 to continue to assess whether or not chlamydial antigens are secreted in a polarized direction. Primary antibodies (ABY) directed against chlamydia specific histones will serve as a control marker for antigens which do not escape the inclusion and mABY directed against Class 1 molecules will serve as a marker for the basolateral domain. Digitized fluorescence microscopy of duplicate samples will give a more quantitative assessment of the pattern of directed antigen secretion. In Specific Aim 3, we shall investigate if chlamydial surface antigen secretion is modulated by estrogen and progesterone. Since the trans- golgi excytosis inhibitor Brefeldin A reduces escape of LPS and MOMP from inclusions, chlamydial antigens may exploit the secretory pathway to reach the infected cell surface. In Specific Aim 4, we shall analyze the pathway of chlamydial antigen sorting by co-localizing escaped chlamydial antigen with the golgi markers beta-COP, p58 and C6-NBD-ceramide. Finally, to begin to examine immune control of chlamydial growth, we have established an in vitro co-cultivation model to assess host effector PMN and T cell responses to infected human epithelial cells.