Interaction of Neisseria gonorrhoeae with polarized human endocervical epithelial

描述(申请人提供)：淋病奈瑟氏菌，一种人类的主要病原体，引起一种常见的性传播感染，淋病。虽然淋球菌(GC)会导致男性生殖器组织的症状性炎症，但女性的GC感染通常没有症状。这种沉默的感染允许细菌传播，这可能导致盆腔炎(PID)，这是异位妊娠和不孕不育的主要原因。播散性淋球菌感染(DGI)可引起化脓性关节炎。孕妇可能会将感染传播给新生儿，导致失明。此外，静止性GC感染增加了患者对艾滋病毒的易感性。这些并发症显然使淋病成为一个重大的公共卫生问题，特别是在女性中。我们的长期目标是描绘女性GC感染的细胞和分子机制。 由于缺乏有效的研究模型，阻碍了对女性生殖道GC感染机制基础的研究。以前的研究主要依赖于非极化的上皮细胞。由于女性生殖道表面被一层极化的上皮细胞保护，这些数据只能适用于临床感染，因为GC与极化和非极化的上皮细胞以相同的方式相互作用。在这项建议中要检验的中心假设是，GC与极化上皮细胞的相互作用与GC与非极化细胞的相互作用显著不同。我们基于强大的初步数据提出了这一假说，该数据表明，肌动蛋白细胞骨架被极化上皮细胞顶端表面的GC贴壁部位排除在外，而不是像在非极化细胞中那样被招募到贴壁部位。 本研究的目的是明确GC与极化和非极化宫颈上皮细胞相互作用的差异，并探讨GC与极化细胞的独特相互作用与GC感染之间的机制联系。为了达到这一目标，我们将使用极化的原代和病毒永生化的人宫颈上皮细胞来追求两个目标：1)检测GC与极化的宫颈上皮细胞相互作用的性质；2)研究GC与极化的宫颈上皮细胞的独特相互作用如何导致GC感染。 极化的原代和病毒永生化的宫颈上皮细胞模型将使我们能够在模拟女性生殖道体内条件的条件下检查GC-上皮细胞的相互作用，并发现女性GC感染的新机制。我们强有力的初步研究表明，拟议的研究有潜力扩大现有的GC感染范例。从拟议的研究中获得的新的机制知识将有助于为淋病和其他性传播感染的干预和治疗制定新的战略。

DESCRIPTION (provided by applicant): Neisseria gonorrhoeae, an obligate human pathogen, causes a common sexually transmitted infection, gonorrhea. While gonococci (GC) cause symptomatic inflammation of the genital tissue in men, GC infections in women are often asymptomatic. The silent infection allows the bacteria to disseminate, which can lead to pelvic inflammatory disease (PID), a leading cause of ectopic pregnancy and infertility. Disseminated gonococcal infection (DGI) may give rise to septic arthritis. Pregnant women can transmit the infection to newborns, causing blindness. Furthermore, silent GC infections increase the susceptibility of patients to HIV. These complications clearly make gonorrhea a significant public health problem, particularly in women. Our long-term goal is to delineate the cellular and molecular mechanisms underlying GC infections in women. The research on the mechanistic basis of GC infection in the female reproductive tract has been hindered by a lack of effective research models. Previous studies have mainly relied on nonpolarized epithelial cells. Because the surface of the female reproductive tract is protected by a layer of polarized epithelial cells, these data can only be applicable to clinical infections f GC interacts with polarized and nonpolarized epithelial cells in the same manner. The central hypothesis to be tested in this proposal is that GC interactions with polarized epithelial cells differ significantly from GC interactions with nonpolarized cells. We have formulated this hypothesis based on our strong preliminary data that show that the actin cytoskeleton is excluded from GC adherent sites at the apical surface of polarized epithelial cells, rather than being recruited to the adherent sites as seen in nonpolarized cells. The goal of this proposal is to define the differences between GC interactions with polarized and nonpolarized endocervical epithelial cells and to investigate the mechanistic links between the unique interaction of GC with the polarized cells and GC infection. To reach this goal, we will use polarized primary and virally immortalized human endocervical epithelial cells to pursue two aims: 1) to examine the nature of GC interactions with polarized endocervical epithelial cells and 2) to investigate how the unique interactions of GC with polarized endocervical epithelia lead to GC infection. Polarized primary and virally immortalized endocervical epithelial cell models will enable us to examine GC-epithelia interactions under the conditions that mimic the in vivo conditions of the female reproductive tract and to discover novel mechanisms that are specific for GC infections in women. Our strong preliminary studies demonstrate the potential of the proposed studies to expand the existing paradigms of GC infections. New mechanistic knowledge derived from the proposed studies will help to generate new strategies for interventions and therapies for gonorrhea and other sexually transmitted infections