Identifying NKT cell lipids of Chlamydia trachomatis and C. muridarum

描述（由申请人提供）：C。沙眼是最常见的可报告的性传播感染（STI），并且每年在美国造成> 100万例病例，在全世界造成约9200万例病例。生殖器感染可导致女性生殖器官的免疫介导的损害和严重的生殖残疾，包括可导致慢性盆腔疼痛、异位妊娠和不孕症的盆腔炎。每年约有8%的女性发生PID，再次感染后，这种风险显著增加40-70%。再感染率约为13%，发生在6个月内。从经济角度来看，治疗衣原体性传播感染和治疗PID每年花费美国卫生保健系统数十亿美元。该项目研究了发展PID/不孕症的潜在促成因素，其结果可用于 用于开发预防PID/不孕症的治疗方法，并最终降低治疗成本。C.沙眼生殖道感染（C. muridarum）用于揭示发展PID/输卵管不孕症的潜在机制，并且被称为上生殖道病理学（UGTP）。我们的体外和体内研究表明，感染C.正如其他人所证实的那样。众所周知，iNKT细胞调节免疫应答的结果，并且可能在针对衣原体的免疫应答中发挥作用。为了了解iNKT细胞如何调节抗衣原体免疫，即帮助消除辐射或促进女性生殖道功能障碍，需要鉴定CD 1d结合糖脂。我们以前的数据显示，糖脂抗原包含在衣原体激活I型（iNKT）和II型NKT细胞在体外使用无细胞的CD 1d抗原呈递试验。尚未鉴定出任何使衣原体在NKT细胞生物学研究中独特的微生物的II型微生物抗原。脂质对于专性细胞内披衣细胞的存活是重要的，所述细胞内披衣细胞合成少量脂质并将来自宿主细胞的其它脂质并入其细胞壁。CD 1d降解是这些细菌的免疫逃避策略，这并不奇怪。具体目标1，我们建议确定CD 1d结合抗原内的衣原体激活NKT细胞在合作博士威廉Hildebrand，在分离CD 1d结合糖脂的专家。在具体目标2中，我们将验证哪些糖脂激活 iNKT细胞，并通过使用流式细胞术和ELISA测定对来自衣原体生殖器感染后缺乏iNKT细胞（C57 BL/6 Bkgd上的CD 1d-/-）的小鼠的匀浆组织进行体内测量细菌负荷、Th 1细胞数量、细胞因子和趋化因子分泌以及UGTP，证明活化的iNKT细胞调节衣原体生殖器感染结果的程度。了解iNKT细胞如何影响衣原体生殖器感染是NKT细胞生物学的一个重要问题，将与NKT细胞生物学专家Mitchell Kronenberg博士合作研究。

DESCRIPTION (provided by applicant): C. trachomatis is the most common reportable sexually transmitted infection (STI) and is responsible for > 1 million cases in the U.S. and approximately 92 million cases worldwide each year. Genital infection can lead to immune-mediated damage of the female reproductive organs and serious reproductive disability, including pelvic inflammatory disease (PID) that can result in chronic pelvic pain, ectopic pregnancy and infertility. Approximately 8% of females annually develop PID and this risk significantly increases by 40-70% following re- infection. The reinfection rate is approximately 13% and occurs within 6 months. Viewed in economic terms, treatment of chlamydial STIs and treatment of PID costs the US health care system billions of dollars annually. This project examines a potential contributing factor for developing PID/infertility and the results can be used for development of therapeutics to prevent PID/infertility and ultimately reduce treatment costs. The mouse model of C. trachomatis genital infection (C. muridarum) is used to reveal the underlying mechanism(s) for developing PID/tubal infertility and is called upper genital tract pathology (UGTP). Our in vitro and in vivo studies show that iNKT are activated by infection with C. muridarum as others have confirmed. It is well known that iNKT cells modulate the outcome of immune responses and likely play a role in the immune response against Chlamydia. In order to understand how iNKT cells modulate anti-chlamydial immune, ie aid in eradiation or contribute to female reproductive tract dysfunction, the identity of CD1d-binding glycolipids are needed. Our previous data revealed that glycolipid antigens contained within chlamydiae activate type I (iNKT) and type II NKT cells in vitro using a cell-free CD1d antigen presentation assay. A type II microbial antigen has not yet been identified for any microbe making Chlamydia unique for the study of NKT cell biology. Lipids are important for survival of the obligate intracellular chlamydiae which synthesize a few lipids and incorporate others into their cell wall from host cells. It is not surprising that CD1d degradation is a strategy for immune evasion of these bacteria. Specific Aim 1, we propose to identify CD1d-binding antigens contained within chlamydiae that activate NKT cells in collaboration with Dr. William Hildebrand, an expert in isolating CD1d- binding glycolipids. In Specific Aim 2, we will validate which glycolipids activate iNKT cells in vivo and demonstrate the extent to which activated iNKT cells modulate the outcome of chlamydial genital infection by measuring bacterial burden, Th1 cell numbers, cytokine and chemokine secretion and UGTP in vivo using flow cytometry and ELISA assays on homogenized tissue from mice that lack iNKT cells (CD1d-/- on C57BL/6 Bkgd) after chlamydial genital infection. Understanding how iNKT cells influence chlamydial genital infection is an important question of NKT cell biology and will be studied in collaboration with Dr. Mitchell Kronenberg, an expert in NKT cell biology.