Role of Toll-Like Receptors in Bacterial Keratitis

Toll 样受体在细菌性角膜炎中的作用

• 批准号: 8206825
• 负责人: LINDA D HAZLETT
• 金额: $ 36.12万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8206825
• 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

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.

项目总结/摘要 铜绿假单胞菌是一种常见的机会致病菌 与细菌性角膜炎相关，特别是在长期配戴接触透镜的使用者中。我们 目的是确定细菌性角膜炎发展的分子机制， 特别是Toll样受体（TLR）4的作用，目前仍然存在 在眼睛里探索不足。我们假设TLR 4在先天性免疫应答中起着关键作用， 免疫反应的铜绿假单胞菌在角膜和激活和信号 通过这种TLR将调节疾病的结果。提出了三个具体目标：1） 测试TLR 4调节感染者中炎性血管生成的假设。 角膜2)为了验证TLR 4调节感染角膜细胞凋亡的假设。 3)为了验证TLR 4调节感染者体内抗菌肽的假设， 角膜实验将包括使用诸如真实的时间RT-PCR、短时间RT-PCR和实时荧光定量PCR等技术。 干扰RNA（siRNA）以及平板计数、酶联免疫吸附试验 （ELISA）、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梯状化、NF B功能测定 和Western印迹。我们的长期目标是了解 细菌和小鼠角膜中的免疫反应， 可以为人类患者开发抗角膜炎的疗法。因为在美国 仅在美国，微生物性角膜炎的发病率为每年25，000 - 30，000例， 治疗费用估计为1500万至3000万美元，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价值。 冲击