STUDIES OF CHLAMYDIA TRACHOMATIS

沙眼衣原体的研究

• 批准号: 6235018
• 负责人: Priscilla B. Wyrick
• 金额: $ 18.22万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 1998-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6235018
• 关键词: Chlamydia trachomatis; antigen presentation; apical membrane; bacterial antigens; bacterial proteins; basolateral membrane; brefeldin A; cellular polarity; cooperative study; endometrium; epithelium; fluorescence microscopy; glycolipids; histocompatibility antigens; host organism interaction; human tissue; immunoelectron microscopy; inclusion body; lipopolysaccharides; membrane proteins; microorganism immunology; secretion; stress proteins; surface antigens; tissue /cell culture

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.

C.沙眼是细菌性传播疾病的主要原因， 在美国传播的感染，并负责25-50%的 估计每年有100万例盆腔炎。 后遗症包括宫外孕和不孕。 为了开始解剖 破坏性与保护性反应，以及确定潜在的 我们的总体目标是了解细胞， 感染衣原体抗原分选和分泌的分子生物学 极化的人子宫内膜上皮细胞。 我们已经检测到衣原体LPS和主要外膜的逃逸 蛋白质（MOMP）的感染细胞的表面之前， 在发育周期结束时释放衣原体。 做 衣原体应激反应蛋白也逃逸到受感染的细胞中 表面：这些抗原从包涵体中通过什么途径， 是宿主免疫反应感知的表面暴露抗原： 我们将使用包埋后免疫电子技术 对Lowicryl处理的样品进行显微镜检查，以确定特定目标1中是否 C.沙眼血清型E GroEL和DnaK也分泌到表面 青霉素和干扰素引起的 持续感染的人子宫内膜上皮细胞， 目的2：继续评估衣原体抗原是否 向两极分化的方向分泌。 一抗（ABY） 针对衣原体特异性组蛋白的抗体将作为对照标记， 不逃避包涵体的抗原和针对 1类分子将用作基底外侧结构域的标志物。 重复样品的数字化荧光显微镜将提供更多 定向抗原分泌模式的定量评估。 在 具体目标3，我们将研究衣原体表面抗原 分泌受雌激素和孕激素调节。 自从跨- 高尔基体胞吞抑制剂Brefeldin A减少LPS和MOMP从 包涵体，衣原体抗原可以利用分泌途径到达 被感染的细胞表面 在具体目标4中，我们将分析 共定位逃逸衣原体的衣原体抗原分选途径 抗原与高尔基体标志物β-COP、p58和C6-NBD-神经酰胺。 最后，为了开始检查衣原体生长的免疫控制，我们 建立了体外共培养模型，以评估宿主效应PMN 和T细胞对感染的人类上皮细胞的反应。