T CELL RECEPTORS/SUSCEPTIBILITY TO RHEUMATOID ARTHRITIS

类风湿性关节炎（RA）的易感性在遗传上与 HLA II类抗原，称为DR4。 基于DNA序列分析，DR4可以 可细分为几个亚型（例如， Dw4、Dw10、Dwl3、Dwl4、Dwl5） 它们彼此仅相差1 - 4个氨基酸。 然而，只有一些 这些亚型与RA相关（即，Dw4、Dw14、Dw15）。 基于 最近在小鼠中的研究表明，II类抗原可以 影响T细胞受体（TCR）库，可以提出， DR4亚型与RA易感性的不同关联是由于 它们对TCR库的不同影响。 这个项目的总体目标是 该项目旨在深入了解HLA II类抗原在 确定TCR库。 特别是，这些研究将侧重于 DR4的各种亚群以及那些也显示 与RA易感性相关。 这些研究将包括 分析来自30名受试者的外周血淋巴细胞的TCR库， 选择已知HLA类型的个体。 TCR剧目将是 使用V β特异性单克隆抗体通过免疫荧光分析 以及使用Valpha和V β特异性引物的定量PCR。 在 此外，还将进行研究以确定不同的TCR是否 用于识别DR4的不同亚型并定义 这是区别他们的特征。 这将通过 分析同种异体反应性免疫应答中单个个体的TCR库， 对用各种DR 4亚型等位基因转染的人B细胞的应答。 此外，研究了HLA II类基因对TCR选择的影响。 将通过确定对DR4亚型应答的所有谱来评估 这些谱系如何随应答者的HLA类型而变化。

Susceptibility to rheumatoid arthritis (RA) is genetically linked to the HLA class II antigen known as DR4. Based on DNA sequence analysis, DR4 can be subdivided into several subtypes (eg. Dw4, Dw10, Dwl3, Dwl4, Dwl5) which differ from each other by only 1-4 amino acids. However, only some of these subtypes are associated with RA (ie. Dw4, Dw14, Dw15). Based on recent studies in mice which demonstrate that class II antigens can influence the T cell receptor (TCR) repertoire, it may be proposed that the differing association of DR4 subtypes with susceptibility to RA is due to their differing effects on the TCR repertoire. The overall goal of this project is to gain insight into the role of HLA class II antigens in determining the TCR repertoire. In particular, these studies will focus on the various subsets of DR4 and those DR1 and DR6 alleles which also display association with susceptibility to RA. The studies will include an analysis of the TCR repertoire of peripheral blood lymphocytes from selected individuals of known HLA type. The TCR repertoire will be analyzed by immunofluorescence using Vbeta-specific monoclonal antibodies and, quantitative PCR using Valpha and Vbeta-specific primers. In addition, studies will be performed to determine whether distinct TCR are used to recognize the different subtypes of DR4 and to define the characteristics that distinguish them. This will be accomplished by analyzing the TCR repertoire of a single individual in an alloreactive response to human B cells transfected with the various DR4 subtype alleles. Furthermore, the influence of HLA class II genes on selection of the TCR repertoire in responses to the DR4 subtypes will be assessed by determining how these repertoires vary with the HLA type of the responder.