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: 3425849
• 负责人: BYOUNG S KWON
• 金额: $ 3.8万
• 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 1994-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3425849
• 关键词: T cell receptor; blood; denaturing gradient gel electrophoresis; human subject; immunogenetics; immunologic techniques; leukocyte activation /transformation; method development; molecular biology; 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.

激活的T细胞似乎是 颞下颌关节（TMJ）类风湿关节炎（RA）。 机制 负责T细胞激活的原因尚不清楚。 如果t 根据常规机制激活细胞，然后是特定的 应通过T细胞抗原受体识别免疫原 （TCR）。 没有定义免疫原或涉及的TCR 在分子水平。 我们假设TCR的主要形式 可以在体内激活的TMJ T中找到可变序列 淋巴细胞。 该提案的主要目的是分析 多样性TCR alpha \ beta V-Region核苷酸序列由In表示 体内激活的T细胞，并表征任何任何的核苷酸序列 主要受体。 为此，我们将选择六个合适的TMJ RA 我们可以从中获得体内活化滑膜的患者。 我们 将开发一种新颖的测定法，结合了“连接器锚定” PCR（LA-PCR） 并贬低梯度凝胶电泳（DGGE）进行定量 估计TCR V区序列的多样性。 这些细胞将是 由LA-PCR Plus DGGE分析。 确定的主要受体物种 凝胶将被克隆并测序。 该测定法的开发将 允许解决克隆性，并且可能需要 确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区域将在A中表达 重组系统和特异性的单克隆抗体将是 生成。 识别主要的T细胞克隆和发展 单克隆抗体可能直接导致重要的医学和科学 申请。 对主要TCR特异的单克隆抗体可能 被证明是诊断和监测疾病的新测定法 活动，并有可能在特定的免疫疗法中。