IMMUNE MECHANISMS OF OCULAR INFLAMMATORY DISEASE

眼部炎症疾病的免疫机制

• 批准号: 2019354
• 负责人: Lynn K Gordon
• 金额: $ 4.47万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-03-01 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2019354
• 关键词: SDS polyacrylamide gel electrophoresis; affinity chromatography; antigen antibody reaction; autoantibody; autoantigens; biomarker; clinical research; cross immunity; enzyme linked immunosorbent assay; family genetics; human subject; immunocytochemistry; immunofluorescence technique; immunoprecipitation; ion exchange chromatography; longitudinal human study; molecular cloning; molecular pathology; monoclonal antibody; nucleic acid sequence; pathologic process; polymerase chain reaction; ulcerative colitis; uveitis; western blottings

The immediate goal of the candidate is to develop scientific expertise in molecular immunology and ocular biochemistry as applied to the definition of pathogenic autoantigens. This will lead to the long-term goal of an independent research program on the pathogenesis of ocular inflammatory diseases. The candidate has a strong background in clinical ophthalmology with a demonstrated interest in ocular inflammation. Past graduate school success in basic research is evidence of her scientific abilities. However, the lag time between graduate school and re-entry into basic research demands a period of mentored scientific retraining. UCLA will provide an outstanding scientific environment that will allow the candidate to develop current expertise in molecular biology and immunology. The health-relatedness of the project is that it specifically attempts to define candidate antigens that drive certain endogenous human inflammatory diseases of the eye, which can be used as the basis for novel diagnostic tests and therapeutic interventions. Although many immune-mediated inflammatory diseases are thought to result from activated CD4 T cells, B cell activation and clonal expansion is expected to occur in tandem. These selected B cells have the same antigenic specificity of the pathogenic T cell, and their antibodies offer an important tool to characterize the target antigen driving the immune response. The subject of this proposal is to use new techniques iii antibody phage display cloning to isolate marker antibodies and candidate antigens for a distinctive type of uveitis that commonly occurs with ulcerative colitis (UC). The first aim is to determine whether a unique marker antibody in UC, pANCA, is associated with inflammatory uveitis. This will be tested by ELISA and immunofluorescence screening of human sera for the distinct UC form of pANCA. The correlation between disease activity and level of autoantibody will also be determined. The second aim is to identify uveal antigens that react with 5-3, a human recombinant monoclonal antibody to the pANCA antigen. If such antigens are identified, they will be characterized by protein biochemistry, or if necessary, by molecular cloning. Disease association of the antigen will be tested by characterizing the B and T cell responses to the antigen. The third aim is to develop a comprehensive library of uveal antigens reactive with the UC antibody response. Biochemical and immunologic characterization of these antigens will be performed following the experimental model of the preceding aim.

候选人的直接目标是发展科学专业知识 在分子免疫学和眼科生物化学中 病原自身抗原的定义。这将导致长期 有关眼发病机理的独立研究计划的目标 炎症性疾病。候选人的临床背景很强 眼科具有对眼部炎症的兴趣。过去的 研究生院基础研究成功是她科学的证据 能力。但是，研究生院与重新进入之间的滞后时间 进入基础研究需要一段经过指导的科学再培训。 加州大学洛杉矶分校将提供一个杰出的科学环境，将允许 发展分子生物学专业知识的候选人和 免疫学。该项目的健康相关性是 尝试定义驱动某些内源人的候选抗原 眼睛的炎症性疾病，可以用作 新颖的诊断测试和治疗干预措施。 尽管人们认为许多免疫介导的炎症性疾病会导致 从活化的CD4 T细胞中，B细胞激活和克隆膨胀为 预计将出现在同时。这些选择的B单元具有相同的 病原T细胞的抗原特异性及其抗体 提供一个重要的工具来表征目标抗原驱动的 免疫反应。该提议的主题是使用新技术 III抗体噬菌体显示克隆以分离标记抗体和 通常发生的独特类型的葡萄膜炎的候选抗原 溃疡性结肠炎（UC）。第一个目的是确定是否 UC中的独特标记抗体与炎症有关 葡萄膜炎。这将通过ELISA和免疫荧光筛查测试 人类血清的独特UC形式的panca。之间的相关性 还将确定疾病活动和自身抗体的水平。这 第二个目的是鉴定与人类反应的紫菜抗原 重组单克隆抗体的panca抗原。如果这样的抗原 被鉴定出来，它们的特征是蛋白质生物化学或 如有必要，通过分子克隆。抗原疾病关联 将通过表征B和T细胞对b和t细胞的反应来测试 抗原。第三个目的是开发一个蛋黄的全面图书馆 抗原用UC抗体反应反应。生化和 将进行这些抗原的免疫特征 遵循前一个目标的实验模型。