VIRULENCE FACTORS OF CHLAMYDIAE

在美国和全世界，沙眼衣原体D-K血清型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、宫外孕和不孕症。衣原体疾病是潜伏的，它们构成了重大的初级、二级和三级健康问题，其中妇女承担着特别的负担，因为她们增加了不良生殖后果的风险。25年来，这个实验室的目标一直是试图了解衣原体在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。这项建议是对关键的附着/进入步骤的持续努力的继续，该步骤是衣原体感染的上皮细胞中触发中性粒细胞趋化的信号，因为对持续的衣原体抗原的长期炎症反应被认为是造成损害和后遗症的原因，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被整合到脂质体中，以确定HSP70在进入体内的作用，在目标2中，帮助识别与新发现的雌激素反应受体辅助蛋白一起发挥功能的受体。同样在目标2中，将建立猪沙眼衣原体S45分离株-猪生殖器组织感染模型，以剖析进入和中性粒细胞信号转导的激素调节(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和持续感染非侵袭性、无症状血清E的极化HeLa细胞的趋化作用与侵袭性、有症状的血清L2。最后，在目标4中，冷冻电子显微镜和密度梯度将被用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出的小泡来发生。

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.