VIRULENCE FACTORS OF CHLAMYDIAE

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，该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和分子后果，因为大多数衣原体输卵管疾病似乎是由慢性亚临床持续感染引起的。 该提议是理解关键附着/进入步骤、衣原体感染的上皮细胞中触发中性粒细胞趋化性的信号的持续努力的延续，因为对持久性衣原体抗原的长期炎症反应被认为是造成损伤和后遗症的原因，以及中性粒细胞进入和信号传导的激素调节。 在目标 1 中，衣原体包膜相关的 hsp70 及其辅助伴侣蛋白 GrpE 和 DnaJ 将与已知和可疑的粘附素一起掺入脂质体中，以定义 hsp70 在进入中的作用，在目标 2 中，帮助识别与新识别的雌激素响应受体辅助蛋白一起发挥作用的受体。 同样在目标 2 中，将开发猪沙眼衣原体 S45 分离株 - 猪生殖组织感染模型，以剖析激素对进入和中性粒细胞信号传导的调节（目标 3）。 在目标 3 中，将比较衣原体和趋化因子信号触发中性粒细胞对正常且持续感染非侵入性、无症状血清型 E 和侵入性、有症状血清型 L2 的极化 HeLa 细胞的趋化性。 最后，在目标 4 中，将使用冷冻电子显微镜和密度梯度来证明衣原体抗原的分泌和运输可以通过从衣原体包涵体中夹断的囊泡进行。

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 their increased risk of adverse reproductive consequences. The goal of this 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 and molecular consequences of persistent infection, since the majority of chlamydial tubal disease appears to result from chronic subclinical, persistent infection. This proposal is a continuation of on-going efforts to understand the crucial attachment/entry steps, the signals in chlamydiae-infected epithelial cells which trigger neutrophil chemotaxis--since a prolonged inflammatory response to persistent chlamydial antigens is believed to be responsible for the damage and sequelae, and hormone modulation of entry and signaling of neutrophils. In Aim 1, the chlamydial envelope-associated hsp70 and its co- chaperonins GrpE and DnaJ will be incorporated into liposomes, along with known and suspected adhesins, to define the role of hsp70 in entry and, in Aim 2, help identify the receptor which functions with newly identified, estrogen-responsive receptor accessory proteins. Also in Aim 2, the swine C. trachomatis S45 isolate- swine genital tissue model of infection will be developed to dissect hormone modulation of entry and neutrophil signaling (Aim 3). In Aim 3, a comparison will be made of chlamydial and chemokine signals triggering neutrophil chemotaxis to polarized HeLa cells normally and persistently infected with non-invasive, asymptomatic serovar E versus invasive, symptomatic serovar L2. Finally, in Aim 4, cryo-electron microscopy and density gradients will be used to show that chlamydial antigen secretion and trafficking can occur via vesicles pinched off from the chlamydial inclusion.