Virulence Factors of Chlamydiae

衣原体的毒力因子

• 批准号: 7081423
• 负责人: Priscilla B. Wyrick
• 金额: $ 41.26万
• 依托单位: EAST TENNESSEE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7081423
• 关键词: Chlamydia trachomatis; bacteria characteristic; bacteria infection mechanism; bacterial antigens; cell adhesion; cell line; chlamydial disease; endoplasmic reticulum; epithelium; estrogen receptors; flow cytometry; membrane fusion; reproductive system infection; small interfering RNA; tissue /cell culture; virulence

DESCRIPTION (provided by applicant): Chlamydia trachomatis serovars D-K are the most common cause, in the USA and worldwide, of bacterially acquired sexually transmitted diseases and their sequelae, including prostatitis, epididymitis, pelvic inflammatory disease, ectopic pregnancy and sterility. Chlamydial diseases are insidious and they constitute significant primary, secondary and tertiary health concerns in which women bear a special burden because of increased risk of adverse reproductive consequences. The goal of our laboratory for 25 years has been to try to understand the basic biology of Chlamydial growth in its host epithelial cell in order to understand the infectious process and to permit dissection of the cellular/molecular consequences of persistent infection, since the majority of Chlamydial tubal disease appears to result from subclinical, persistent infection. This proposal is a continuation of on-going efforts to understand the crucial attachment steps, which are modulated by female hormones in vitro. In Specific Aim 1, we shall determine whether or not estrogen receptors (ER) are involved in serovar E attachment to human endometrial epithelial cells or, alternatively, if the estrogen-induced physiological effects on the host cell result in display of other receptors. Specific labeled-EB binding assays to ER+ cell lines will be used in addition to BiaCore, FACS, and microscopy analyses and coupled with siRNA technology to knockdown surface ER amounts to see if reduced EB binding does occur. In Specific Aim 2, we shall monitor, by microscopy and gradient vesicle isolation, mechanisms of serovar E outer membrane bleb antigen escape through the inclusion membrane as well as from inclusion membrane everted vesicles and determine trafficking, including endoplasmic reticulum membrane fusion potential for MHC Class I loading. Finally, for Specific Aim 3, we have isolated Chlamydial projections (small scale) and, after bulk collection and generation of antibodies, plan to characterize these surface structures; the obvious question is whether or not the projections are the Chlamydial Type III secretion injectisome-equivalent.

描述（由申请人提供）：沙眼衣原体 D-K 血清型是美国和全世界细菌性性传播疾病及其后遗症的最常见原因，包括前列腺炎、附睾炎、盆腔炎、宫外孕和不孕症。衣原体疾病是隐匿性的，构成重大的一级、二级和三级健康问题，妇女由于不良生殖后果的风险增加而承受着特殊的负担。 25 年来，我们实验室的目标一直是尝试了解衣原体在其宿主上皮细胞中生长的基本生物学，以便了解感染过程并剖析持续感染的细胞/分子后果，因为大多数衣原体输卵管疾病似乎是由亚临床持续感染引起的。该提案是理解关键依恋步骤的持续努力的延续，这些步骤是由体外雌性激素调节的。在具体目标 1 中，我们将确定雌激素受体 (ER) 是否参与血清 E 与人子宫内膜上皮细胞的附着，或者雌激素诱导的对宿主细胞的生理作用是否导致其他受体的展示。除了 BiaCore、FACS 和显微镜分析之外，还将使用针对 ER+ 细胞系的特异性标记 EB 结合测定，并结合 siRNA 技术来敲低表面 ER 量，以查看是否确实发生了 EB 结合减少。 在具体目标 2 中，我们将通过显微镜和梯度囊泡分离监测血清型 E 外膜泡抗原通过包涵膜以及包涵膜外翻囊泡逃逸的机制，并确定运输，包括 MHC I 类负载的内质网膜融合潜力。 最后，对于特定目标 3，我们分离了衣原体投影（小规模），并在批量收集和生成抗体后，计划表征这些表面结构；明显的问题是预测是否与 III 型衣原体分泌注射体等效。