NEW ASSAY FOR TCR DIVERSITY IN TMJ RHEUMATOID ARTHRITIS

颞下颌关节类风湿关节炎 TCR 多样性的新检测

• 批准号: 2131418
• 负责人: BYOUNG S KWON
• 金额: $ 3.82万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1995-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2131418
• 关键词: T cell receptor; blood; denaturing gradient gel electrophoresis; human subject; immunogenetics; immunologic techniques; leukocyte activation /transformation; method development; molecular biology; nucleic acid sequence; point mutation; polymerase chain reaction; rheumatoid arthritis; synovial fluid; temporomandibular joint

The activated T-cells appear to be a central element in the pathogenesis of temporomandibular joint (TMJ) rheumatoid arthritis (RA). The mechanisms responsible for the activation of the T cells are not known. If the T cells are activated according to conventional mechanisms, then specific immunogen(s) should be recognized through the T-cell antigen receptor (TCR). Neither the immunogen(s), nor the TCR;s involved have been defined at the molecular level. We hypothesize that a predominant form of TCR variable sequence can be found among the in vivo activated TMJ T lymphocytes. The primary objective of this proposal is to analyze the diversity TCR alpha/beta V-region nucleotide sequences expressed by the in vivo activated T cells, and characterize the nucleotide sequences of any predominant receptors. To this end, we will select six suitable TMJ RA patients from which we can obtain in vivo activated synovial T- cells. We will develop a novel assay which combines "linker-anchored" PCR (LA-PCR) and Denaturing Gradient Gel Electrophoresis (DGGE) to quantitatively estimate the diversity of TCR V-region sequences. These cells will be analyzed by the LA-PCR plus DGGE. Predominant receptor species identified on the gel will be cloned and sequenced. Development of this assay would allow resolution of the clonality, and might in the future be needed to identify patient-specific pathogenic T cell clones. As a future effort, unique regions of these predominant receptors will be expressed in a recombinant system, and monoclonal antibodies specific for them will be generated. Identification of predominant T cell clones, and development of monoclonal antibody could lead directly to important medical and scientific applications. Monoclonal antibodies specific for predominant TCR might prove useful as new assays for diagnosis and for monitoring disease activity, and potentially in specific immunotherapy.

活化的T细胞似乎是发病机理的中心元素 颞下颌关节（TMJ）类风湿关节炎（RA）。 这 导致T细胞激活的机制尚不清楚。 如果根据常规机制激活T细胞，则 应通过T细胞抗原识别特定的免疫原 受体（TCR）。 免疫原子和所涉及的TCR都没有 在分子水平上定义。 我们假设一个主要的 在体内激活中可以找到TCR变量序列的形式 TMJ T淋巴细胞。 该提案的主要目的是分析 多样性TCRα/βV-区域核苷酸序列由 体内激活的T细胞，并表征核苷酸序列 任何主要受体。 为此，我们将选择六个合适的 我们可以从中获得体内激活滑膜T-的TMJ RA患者 细胞。 我们将开发一种新颖的测定法，结合了“连接器锚定” PCR（LA-PCR）和剥落梯度凝胶电泳（DGGE）至 定量估计TCR V区序列的多样性。 这些 细胞将通过LA-PCR Plus DGGE分析。 主要受体 在凝胶上鉴定的物种将被克隆并测序。 发展 该测定法可以解决克隆性，并可能在 需要未来以鉴定患者特异性的病原T细胞克隆。 作为未来的努力，这些主要受体的独特地区将 在重组系统中表达，单克隆抗体特异性 为他们生成。 鉴定主要的T细胞克隆， 单克隆抗体的开发可能直接导致重要 医学和科学应用。 特有的单克隆抗体 主要的TCR可能被证明是诊断的新测定法和 监测疾病活动，并有可能在特定的免疫疗法中进行监测。