The Role of Toll-Like Receptors in the Pathogenesis of Dry Eye

Toll 样受体在干眼发病机制中的作用

• 批准号: 7915425
• 负责人: RACHEL LEIGH REDFERN
• 金额: $ 13.5万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7915425
• 关键词: Age; Apoptosis; Automobile Driving; Bacteria; Blindness; C57BL/6 Mouse; Cell Line; Cells; Cicatrix; Confocal Microscopy; Cornea; Cytology; Doctor of Philosophy; Dry Eye Syndromes; Epithelial Cells; Family; Flow Cytometry; Gelatinase B; Glycoproteins; Goblet Cells; Human; Immunology; Infection; Inflammation; Inflammatory; Knock-out; Laboratories; Measures; Mentors; Messenger RNA; Modeling; Mus; Optometrist; Pathogenesis; Patients; Pattern; Population; Prevention; Principal Investigator; Production; Public Health; Quality of life; Reaction; Reading; Reporting; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Risk; Role; Sampling; Simian virus 40; Sjogren' s Syndrome; Source; Stimulus; Stress; Symptoms; TLR2 gene; Testing; Therapeutic Intervention; Time; Toll-Like Receptor 2; Toll-like receptors; Training; Training Programs; Tumor Necrosis Factor-alpha; Ulcer; Virus; Work; chemokine; conjunctiva; corneal epithelium; cytokine; experience; eye dryness; fungus; human TNF protein; human subject; impression; insight; irritation; multidisciplinary; novel; ocular surface; programs; protein expression; receptor; receptor expression; response; skills

DESCRIPTION (provided by applicant): The primary objective of the proposed research plan is to provide the Principal Investigator (PI) with the comprehensive didactic and laboratory training necessary to develop into an independent researcher in ocular surface inflammation. The multidisciplinary training plan will be under the guidance of mentors Alison McDermott, PhD and Stephen Pflugfelder, MD. The PI is an Optometrist with a background in corneal immunology and this proposed research program will expand upon the her previous scientific experience through studies to investigate the role of Toll-like receptors (TLRs) in ocular surface inflammation in dry eye syndrome. The activation of TLRs has been shown to stimulate the production of cytokines and chemokines, initiating an inflammatory reaction. The scientific focus of this proposal will investigate the general hypothesis that TLRs stimulate inflammation in dry eye syndrome. The specific aims are: (1) To establish TLR 2, 4, 5 and 9 expression pattern in human conjunctival epithelial cells from subjects with mild, moderate, and severe dry eye, Sjogren's syndrome, and age matched normal subjects. Conjunctival impression cytology samples will be collected from human subjects and TLR mRNA and protein expression will be determined by real time RT-PCR and flow cytometry respectively; (2) To determine the expression and localization pattern of TLR 2, 4, 5 and 9 in primary human conjunctival and corneal epithelial cells and in normal human conjunctival and corneal epithelial cell lines in response to hyperosmolar stress and cytokine treatment by real time RT-PCR and confocal microscopy; (3) To compare the TLR 2, 4, 5 and 9 expression pattern among control C57BL/6 mice with experimental dry eye (EDE) induced by desiccating stimulus C57BL/6 mice. Additionally this specific aim will determine if EDE TLR 2, 4, and 9 deficient mice have reduced ocular surface inflammation as measured by a.) a reduction in MMP-9 and cytokine (IL-1beta and TNF-alpha) mRNA and protein expression and b.) epithelial cell apoptosis and goblet cell loss compared to wild type EDE C57BL/6 mice. These studies will help understand the role of TLRs in the pathogenesis of dry eye while also providing the PI with a training program to develop the skills necessary to become a successful independent investigator.

描述（由申请人提供）：拟议的研究计划的主要目的是向主要研究者（PI）提供必要的全面的教学和实验室培训，以发展为眼部表面炎症中独立的研究人员。多学科培训计划将在导师Alison McDermott博士和医学博士Stephen Pflugfelder的指导下。 PI是具有角膜免疫学背景背景的验光师，该拟议的研究计划将通过研究通过研究来扩展她以前的科学经验，以研究Toll样受体（TLR）在眼部表面炎症中在干眼综合征中的作用。 TLR的激活已被证明可以刺激细胞因子和趋化因子的产生，即 引发炎症反应。该提案的科学重点将研究以下一般假设：TLR刺激干眼综合征的炎症。具体目的是：（1）在人类结膜上皮细胞中建立TLR 2、4、5和9的表达模式，来自轻度，中度和严重的干眼症患者，Sjogren综合征以及年龄匹配的正常受试者。结膜印象细胞学样本将从人类受试者中收集，TLR mRNA和蛋白质表达将分别通过实时RT-PCR和流式细胞仪确定。 （2）确定原代人结膜和角膜上皮细胞中TLR 2、4、5和9的表达和定位模式，以及正常的人类结膜和角膜上皮细胞系，响应于高温应激和细胞因子治疗，并通过实时RT-PCR-PCR-PCR-PCR-PCR-PCR-PCR-PCR-PCR-PCRECAL和共焦显微镜处理； （3）比较对照C57BL/6小鼠之间的TLR 2、4、5和9的表达模式，并与通过干燥刺激C57BL/6小鼠诱导的实验干眼（EDE）进行了比较。此外，该特定目的将确定EDE TLR 2、4和9不足的小鼠是否通过A。测量的眼部表面炎症减少了。 这些研究将有助于了解TLR在干眼症的发病机理中的作用，同时还为PI提供了培训计划，以发展成为成功的独立研究者所必需的技能。