Virulence Factors of Chlamydiae

衣原体的毒力因子

• 批准号: 6913865
• 负责人: Priscilla B. Wyrick
• 金额: $ 33.15万
• 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/6913865
• 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（由申请方提供）：沙眼衣原体血清型D-K是美国和世界范围内细菌获得性性传播疾病及其后遗症（包括前列腺炎、附睾炎、盆腔炎、异位妊娠和不孕症）的最常见原因。衣原体疾病是潜伏性的，它们构成了重大的一级、二级和三级健康问题，妇女承受着特殊的负担，因为它们增加了对生殖产生不利后果的风险。我们实验室25年来的目标一直是试图了解衣原体在其宿主上皮细胞中生长的基本生物学，以了解感染过程，并允许解剖持续感染的细胞/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。这个建议是对正在进行的努力的延续，以了解关键的依恋步骤，这些步骤是由体外女性激素调节的。在具体目标1中，我们将确定雌激素受体（ER）是否参与血清型E对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显示。除BiaCore、FACS和显微镜分析外，还将使用ER+细胞系的特异性标记EB结合试验，并结合siRNA技术来敲低表面ER量，以观察是否确实发生EB结合减少。 在特定目标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体膜以及从包涵体膜外翻囊泡逃逸的机制，并确定运输，包括用于MHC I类负载的内质网膜融合潜力。 最后，对于特定目标3，我们分离了衣原体的突起（小规模），并在大量收集和产生抗体后，计划表征这些表面结构;明显的问题是这些突起是否与衣原体III型分泌物注射体相当。