Role of Toll-Like Receptors in Bacterial Keratitis

Toll 样受体在细菌性角膜炎中的作用

基本信息

批准号：
8386603
负责人：
LINDA D HAZLETT
金额：
$ 34.31万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-01-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8386603
关键词：
Address Antibodies Apoptosis Apoptotic Bacteria Biological Assay Blindness Cells Clinical Cornea Corneal Diseases DNA Data Defensins Development Dinoprostone Disease Disease Outcome Endotoxins Extended-Wear Contact Lenses Eye Goals Healed Health Histopathology Human Immune response Inbred BALB C Mice Incidence Infection Inflammatory Injection of therapeutic agent Interleukin-1 Receptors Keratitis Leukocytes Link Medical Economics Messenger RNA Molecular Molecular Immunology Molecular and Cellular Biology Mus Natural Immunity Organism Pathogenesis Patients Perforation Peroxidases Phenotype Play Process Production Protein Family Pseudomonas Pseudomonas aeruginosa RNA Receptor Signaling Recombinant Proteins Regulation Reverse Transcriptase Polymerase Chain Reaction Role Signal Pathway Signal Transduction Staining method Stains TLR4 gene Techniques Testing Time Toll-like receptors Treatment Cost United States Vascular Endothelial Growth Factors Vision research Western Blotting abstracting angiogenesis antimicrobial peptide bacterial resistance caspase-3 clinical care cyclooxygenase 2 early onset economic impact healing inflammatory modulation insight killings member microbial neutrophil novel pathogen prevent pro-caspase-3 programs research study response toll-like receptor 4

项目摘要

Project Summary/Abstract Pseudomonas aeruginosa (P. aeruginosa) is a common opportunistic organism associated with bacterial keratitis, especially in extended wear contact lens users. Our goal is to determine the molecular mechanisms for development of bacterial keratitis, specifically, the role of Toll-like receptor (TLR) 4, which at present remains underexplored in the eye. We hypothesize that TLR4 plays a critical role in the innate immune response to P. aeruginosa in the cornea and that the activation and signaling through this TLR will regulate disease outcome. Three specific aims are proposed: 1) To test the hypothesis that TLR4 regulates inflammatory angiogenesis in the infected cornea. 2) To test the hypothesis that TLR4 regulates apoptosis in the infected cornea. 3) To test the hypothesis that TLR4 regulates antimicrobial peptides in the infected cornea. Experiments will include use of techniques such as real time RT-PCR, short interfering RNA (siRNA), as well as plate counts, enyme linked immunosorbant assay (ELISA), myeloperoxidase assay (MPO) for neutrophil (PMN) quantitation, dual antibody immunostaining, histopathology, Tunel assay, DNA laddering, NF¿B functional assays and Western blotting. Our long-term objective is to understand the interactions between bacteria and the immune response in the mouse cornea so that non-conventional therapies against keratitis can be developed for human patients. Since in the United States alone the incidence of microbial keratitis is 25,000-30,000 cases annually, with cost of treatment estimated at $15-30 million, and with emerging bacterial resistance, the studies are of relevance to human health and have considerable medical and economic impact.

项目概要/摘要铜绿假单胞菌（P. aeruginosa）是一种常见的机会性微生物与细菌性角膜炎有关，尤其是长期佩戴隐形眼镜的使用者。目标是确定细菌性角膜炎发生的分子机制，具体来说，Toll 样受体 (TLR) 4 的作用目前仍然存在我们发现 TLR4 在眼睛中发挥着关键作用。对角膜中铜绿假单胞菌的免疫反应以及激活和信号传导通过该 TLR 将调节疾病结果。提出了三个具体目标：1）检验 TLR4 调节感染者炎症血管生成的假设 2) 检验 TLR4 调节受感染角膜细胞凋亡的假设。 3) 检验TLR4调节感染者体内抗菌肽的假设实验将包括使用实时 RT-PCR 等技术。干扰 RNA (siRNA)，以及平板计数、酶联免疫吸附测定 (ELISA)、用于中性粒细胞 (PMN) 定量的髓过氧化物酶测定 (MPO)、双抗体免疫染色、组织病理学、Tunel 测定、DNA 阶梯分析、NF¿ B 功能测定我们的长期目标是了解两者之间的相互作用。细菌和小鼠角膜中的免疫反应，因此非常规自美国以来，可以为人类患者开发针对角膜炎的疗法。仅在一个州，每年就有 25,000-30,000 例微生物性角膜炎发生，其中治疗费用估计为 15-3000 万美元，而且随着细菌耐药性的出现，研究与人类健康相关，具有相当大的医学和经济意义影响。