VIRULENCE FACTORS OF CHLAMYDIAE

在美国和世界范围内，D-K型沙眼衣原体是细菌获得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。该实验室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，以便了解感染过程，并允许解剖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。这一建议是对了解关键的附着/进入步骤、衣原体感染的上皮细胞中触发中性粒细胞趋化的信号的持续努力的延续——因为对持续存在的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的激素调节。在Aim 1中，衣原体包膜相关的hsp70及其共伴侣蛋白GrpE和DnaJ将与已知和可疑的粘附素一起纳入脂质体，以确定hsp70在进入中的作用，并在Aim 2中帮助识别与新发现的雌激素应答受体辅助蛋白一起起作用的受体。同样在第二阶段，猪沙眼衣原体S45分离物-猪生殖器组织感染模型将被开发，以解剖进入和中性粒细胞信号的激素调节（第三阶段）。在Aim 3中，将比较衣原体和趋化因子信号触发中性粒细胞趋化到极化HeLa细胞正常和持续感染无创、无症状的血清型E与有创、有症状的血清型L2。最后，在aims 4中，冷冻电子显微镜和密度梯度将用于显示衣原体抗原的分泌和运输可以通过从衣原体包涵体中挤出的囊泡发生。

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.