Polymicrobial Context of Neisseria gonorrhoeae Infection and Mucosal Immune Response

项目概要/摘要 淋病奈瑟菌是一种引起性传播感染 (STI) 的革兰氏阴性细菌 淋病。据估计，全球每年有 7800 万淋病病例，淋病发病率不断增加 由于对所有推荐的抗生素具有耐药性，并且缺乏保护性疫苗，淋病奈瑟菌是一个突出的问题 并对人类健康构成日益严重的威胁。在女性中，淋病奈瑟菌会在子宫颈处形成感染。 引发以中性粒细胞局部募集为特征的炎症反应。活的淋病奈瑟菌 从人类淋病渗出物中回收，表明中性粒细胞不能有效清除感染。 由此产生的持续感染和中性粒细胞炎症循环使得淋病奈瑟菌能够传播 以及上行感染和组织损伤，这是盆腔炎等后遗症的基础， 宫外孕、不孕不育。此外，淋病奈瑟菌感染与流行病学高度相关 与沙眼衣原体合并感染，增加了这些负面后果的可能性。识别 这些重要的性传播感染病原体引起的宫颈感染和炎症的机制对于 寻找新的治疗方法来增强女性的整体健康和生殖健康。我们的知识 促进女性淋病奈瑟菌感染的细菌和宿主条件受到以下因素的阻碍 缺乏包含常驻和招募宿主细胞的人类生殖器感染的稳健模型， 维持女性下生殖道的结构，并纳入生殖器微生物群。克服 针对这个知识缺口，我们将使用我们课题组开发的3D人体初级宫颈阴道仿生系统 单独回答有关女性生殖器感染生物学的基本问题 我们假设宫颈阴道微生物群与易感性有关（细菌性阴道病相关 厌氧菌，群落状态 IV 型）和耐药性（卷曲乳杆菌为主，群落状态类型 - 我）。目标 1 将确定微生物群对宫颈阴道淋病奈瑟菌感染进展的影响 以及对上皮宿主防御的影响。目标 2 将描述微生物群对子宫颈阴道的影响 招募免疫细胞以应对淋病奈瑟菌。目标 3 将定义淋病奈瑟菌合并感染的方式 微生物群中的沙眼衣原体和沙眼衣原体影响病原体负荷和宫颈阴道免疫 回应。这些研究将包括女性的阴道和宫颈内膜细胞及其自体细胞 微生物群以及无与伦比的微生物群，以评估宿主-微生物群相互作用如何影响 感染的进展和结果。这些关于感染过程的研究结果 随之而来的炎症反应和组织完整性可以揭示新的干预措施，以限制细菌负荷和 粘膜上皮损伤，从而减轻这些常见和 使人衰弱的细菌性传播感染。

PROJECT SUMMARY/ABSTRACT Neisseria gonorrhoeae is a Gram-negative bacterium that causes the sexually transmitted infection (STI) gonorrhea. With an estimated 78 million cases of gonorrhea annually worldwide, increasing frequency of resistance to all recommended antibiotics, and the lack of a protective vaccine, N. gonorrhoeae is a prominent and growing threat to human health. In women, N. gonorrhoeae establishes infection at the cervix, where it initiates an inflammatory response characterized by the local recruitment of neutrophils. Viable N. gonorrhoeae are recovered from human gonorrheal exudates, indicating that neutrophils cannot effectively clear infection. The resulting cycle of sustained infection and neutrophilic inflammation enables N. gonorrhoeae transmission as well as ascending infection and tissue damage, which underlie sequelae such as pelvic inflammatory disease, ectopic pregnancy, and infertility. Moreover, infection with N. gonorrhoeae is highly epidemiologically associated with coinfection with Chlamydia trachomatis, enhancing likelihood of these negative consequences. Identifying the mechanisms underlying cervical infection and inflammation by these prominent STI pathogens is critical for finding new therapeutic approaches to enhance women’s overall health and reproductive fitness. Our knowledge of the bacterial and host conditions that facilitate productive N. gonorrhoeae infection in women is hampered by the absence of a robust model for human genital infection that incorporates resident and recruited host cells, maintains the architecture of the lower female genital tract, and incorporates the genital microbiota. To overcome this knowledge gap, we will use the 3D human primary cervicovaginal biomimetic system developed by our group to answer fundamental questions about the biology of female genital infection, alone and in the context of cervicovaginal microbiota that we hypothesize are associated with susceptibility (bacterial vaginosis-associated anaerobes, community state type-IV) and resistance (Lactobacillus crispatus-dominant, community state type- I). Aim 1 will define the influence of the microbiota on the progression of cervicovaginal N. gonorrhoeae infection and effects on epithelial host defenses. Aim 2 will characterize the effect of the microbiota on the cervicovaginal recruitment of immune cells in response to N. gonorrhoeae. Aim 3 will define how coinfection with N. gonorrhoeae and C. trachomatis in the context of the microbiota impacts pathogen burden and cervicovaginal immune response. These studies will include vaginal and endocervical cells from women along with their autologous microbiota, as well as unmatched microbiota, to evaluate how the host-microbiota interaction influences the progression and outcome of infections. The findings arising from these studies about the infection process and ensuing inflammatory response and tissue integrity can reveal new interventions to limit bacterial load and mucosal epithelial damage, thereby mitigating the negative consequences in women of these common and debilitating bacterial STIs.