Virulence Factors of Chlamydiae

衣原体的毒力因子

• 批准号: 7196436
• 负责人: Priscilla B. Wyrick
• 金额: $ 41.27万
• 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/7196436
• 关键词: Antibodies; Antigens; Bacterial Typing; Binding; Biological Assay; Biology; Bulla; Cell Communication; Cell Line; Cells; Cervix Uteri; Chlamydia; Chlamydia Infections; Chlamydia trachomatis; Clinical; Collection; Coupled; Cytoplasm; Development; Disease; Dissection; Ectopic Pregnancy; Endometrial; Endoplasmic Reticulum; Epidemic; Epididymitis; Epithelial Cells; Epithelium; Estrogen Receptors; Estrogens; Female; Generations; Genomics; Glean; Goals; Growth; HIV Infections; Health; Health Care Costs; Healthcare; Hormones; Human; In Vitro; Infection; Infertility; Inflammatory; Label; Laboratories; Learning; Left; MHC Class I Genes; Mammalian Oviducts; Membrane; Membrane Fusion; Microbiology; Microscopy; Molecular; Monitor; Nature; Needles; Neutrophil Infiltration; Pathogenesis; Patients; Pelvic Inflammatory Disease; Personal Communication; Personal Satisfaction; Physiological; Play; Preparation; Process; Receptor Cell; Research; Research Personnel; Risk; Risk Factors; Role; Serological; Sexually Transmitted Diseases; Small Interfering RNA; Sperm Motility; Staging; Sterility; Structure; Surface; Technology; Ursidae Family; Variant; Vesicle; Virulence Factors; Woman; appendage; genital infection; insight; male; pathogen; programs; prostatitis; receptor; reproductive; sperm cell; trafficking; transcriptomics

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）：在美国和世界范围内，沙眼衣原体血清型D-K是细菌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、异位妊娠和不孕症。衣原体疾病具有隐蔽性，它们构成了重大的初级、二级和三级健康问题，妇女因其不利生殖后果的风险增加而承受着特别的负担。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。这一建议是持续不断的努力，以了解关键的依恋步骤，这是由体外雌性激素调节的。在Specific Aim 1中，我们将确定雌激素受体（ER）是否参与血清E附着于人子宫内膜上皮细胞，或者，雌激素诱导的宿主细胞生理效应是否导致其他受体的显示。除了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技术降低表面ER量，以观察EB结合是否减少。在Specific Aim 2中，我们将通过显微镜和梯度囊泡分离来监测血清型E外膜泡抗原通过包涵膜和从包涵膜外翻囊泡逃逸的机制，并确定运输，包括内质网膜融合电位用于MHC I类装载。最后，对于Specific Aim 3，我们分离了衣原体投影（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；显而易见的问题是，这些突出物是否与III型衣原体分泌的注射体相当。