Gonococcal interactions with human polymorphonuclear leukocytes

淋病奈瑟菌（Gc）是一种人类特有的细菌病原体，可引起性传播疾病。 淋病在受感染的个体中，大量的多形核白细胞（PMNs） 被招募到感染部位。然而，所产生的渗出液含有活的生物体， 传播到新的宿主，表明PMN抗菌机制不足以清除淋病， 感染.在体外，PMNs可以杀死一定比例的Gc，但其余的Gc则不能杀死。 生存和扩散。中性粒细胞具有产生活性氧的多亚基NADPH氧化酶 具有杀菌活性的活性氧（ROS）。然而，NADPH氧化酶活性对PMN的杀伤作用不大， 我发现这是因为Gc具有令人惊讶的抑制ROS生成的能力， PMNs。因此，中性粒细胞将其非氧化性抗微生物机制导向Gc。我有 鉴定了两种Gc基因产物，ngol 686和recN，其保护Gc免受PMN的杀伤。这个的目标 建议是1）鉴定可以杀死Gc的非氧化性PMN抗菌因子和2）确定 ngo 1686和recN突变体对PMN杀伤敏感性增加的潜在机制。我会 鉴定对Gc和I具有体外杀菌活性PMN抗微生物因子将确定 Gc是否与这些因子共定位。所有这些研究都将在 Gc与纯化的PMN杀菌因子、PMN颗粒提取物或完整的人PMN组合 从静脉血中分离出来 我已经研究粘膜病原体与宿主细胞的相互作用12年了。在我 博士博士后奖学金。在西北大学史蒂文·塞弗特的实验室里， Gc如何逃避PMN清除，这是Gc发病机制中一个重要但研究较少的方面。在为期一年的K99中 在独立之路奖的第一阶段，我完成了Gc-PMN相互作用的初步研究，奠定了 为本ROO提案中概述的具体目标奠定基础。我将进行这项独立研究， 弗吉尼亚大学健康科学中心的微生物学系， 被任命为终身助理教授，2008年10月1日开始。研究支持 ROO奖将为我的实验室当前和未来的重点领域提供关键基础， 探索对奈瑟菌感染的先天免疫反应。

Neisseria gonorrhoeae (Gc) is a human-specific bacterial pathogen that causes the sexually transmitted disease gonorrhea. In infected individuals, massive numbers of polymorphonuclear leukocytes (PMNs) are recruited to the site of infection. However, the resulting exudate contains viable organisms that are transmitted to new hosts, indicating that PMN antimicrobial mechanisms are insufficient to clear gonorrheal infections. In vitro, PMNs can kill a percentage of Gc with which they are presented, but the remainder survive and proliferate. PMNs possess a multisubunit NADPH oxidase that generates reactive oxygen species (ROS) with bactericidal activity. However, NADPH oxidase activity is dispensable for PMN killing of Gc, and I have found that this is because Gc have the surprising ability to suppress ROS generation in PMNs. As a result, PMNs are directing their non-oxidative antimicrobial machinery against Gc. I have identified two Gc gene products, ngol 686 and recN, that protect Gc from killing by PMNs. The goals of this proposal are 1) to identify the non-oxidative PMN antimicrobial factors that can kill Gc and 2) to define the mechanisms underlying the increased sensitivity of the ngo1686 and recN mutants to PMN killing. I will identify PMN antimicrobial factors that have in vitro bactericidal activity against Gc and I will determine whether Gc colocalize with these factors during exposure to PMNs. All these studies will be conducted with Gc in combination with purified PMN bactericidal factors, PMN granule extracts, or intact human PMNs isolated from venous blood. I have been studying the interplay of mucosal pathogens with host cells for 12 years. During my postdoctoral fellowship in Dr. H. Steven Seifert's laboratory at Northwestern University, I began to address how Gc evade PMN clearance, a critical but poorly studied aspect of Gc pathogenesis. In the one-year K99 phase of the Pathway to Independence award, I completed initial studies of Gc-PMN interactions that laid the groundwork for the specific aims outlined in this ROO proposal. I will conduct this independent research in the Department of Microbiology at the University of Virginia Health Sciences Center, where I have been appointed a tenure-track Assistant Professor with a start date of October 1, 2008. The research supported by the ROO award will provide the critical foundation for my laboratory's current and future area of focus, exploring the innate immune response to Neisserial infection.