Role of Toll-Like Receptors in Bacterial Keratitis

描述（由申请方提供）：铜绿假单胞菌（P. aeruginosa）是一种与细菌性角膜炎相关的常见微生物，尤其是在长期配戴接触透镜的使用者中。在美国，微生物角膜炎的发病率为每年25，000 - 30，000例，治疗费用估计为1500 - 3000万美元，使得该疾病具有相当大的医疗和经济影响。在提出的研究中，我们将检验Toll样受体/白细胞介素-1受体（TLR/IL-IR）及其相关分子（例如，CD 14）在角膜中的初始（先天）免疫应答中起关键作用，并且负责B6和BALB/c小鼠对铜绿假单胞菌感染的不同结果。没有其他实验室正在使用细菌（非无菌）角膜炎模型研究该受体家族在眼睛中的作用。提出了三个目标，将测试假设：1）TLR/IL-1 Rs的表达水平/分布（TLR-4、TLR-9、ST 2和SIGIRR）和相关分子在正常和/或铜绿假单胞菌感染的角膜中，在B6和BALB/c小鼠中上皮和基质中的CD 14、MD 2是不同的; 2）TLR/IL 1-R MyD 88与TRIF信号通路在细菌感染后的易感小鼠与抗性小鼠中优先激活;（3）TLR/IL-1 R及其信号分子在B6和BALB/c小鼠感染角膜中协同调节获得性免疫（Th 1和Th 2应答）。因此，本申请中提出的研究的目的是阐明TLR/IL-1 R在对实验性铜绿假单胞菌角膜感染的先天性和适应性免疫应答中的作用，所述实验性铜绿假单胞菌角膜感染导致角膜穿孔（B6）与愈合（BALB/c）。在拟定的研究中，将使用多种方法，包括使用体内和体外模型来检测TLR表达水平、分布、信号传导和免疫调节特性。 预计这些发现将揭示早期干预和/或治疗铜绿假单胞菌角膜炎的潜在目标。

DESCRIPTION (provided by applicant): Pseudomonas aeruginosa (P. aeruginosa) is a common organism associated with bacterial keratitis, especially in extended wear contact lens users. In the United States, the incidence of microbial keratitis is 25,000-30,000 cases annually with cost of treatment estimated at $15-30 million, making the disease of considerable medical and economic impact. In the studies proposed, we will test the overall hypothesis that Toll-like receptor/interleukin-1 receptor (TLR/IL-IR) and their associated molecules (e.g., CD14) play a critical role in the initial (innate) immune response in the cornea and are responsible for the disparate outcome to P. aeruginosa infection of B6 and BALB/c mice. No other laboratory is studying the role of this family of receptors in the eye using a bacterial (non-sterile) keratitis model. Three aims are proposed that will test the hypotheses that: 1) expression levels/distribution of TLR/IL-1Rs (TLR-4, -9, ST2 and SIGIRR) and related molecules (CD14, MD2) in epithelium and stroma are disparate in B6 and BALB/c mice in normal and/or P. aeruginosa infected cornea; 2) the TLR/IL1-R MyD88 vs. TRIF signaling pathway is preferentially activated in susceptible vs. resistant mice after bacterial infection; and 3) adaptive immunity (Th1 vs. Th2 responsiveness) is regulated disparately by TLR/IL-1Rs and their signaling molecules in the infected cornea of B6 vs. BALB/c mice. Thus, the goal of the studies proposed in this application is to elucidate the role of TLR/IL-1R in the innate and adaptive immune response to experimental P. aeruginosa corneal infection resulting in corneal perforation (B6) vs. healing (BALB/c). In the proposed studies, multiple approaches will be used and include use of both in vivo and in vitro models to test TLR expression levels, distribution, signaling and immunoregulatory properties. It is expected that the findings will reveal potential targets for early intervention and/or treatment of P. aeruginosa keratitis.