HLA AND OTHER GENES AND THE TCR REPERTOIRE

该提案的主要目标是确定影响的例子 单个HLA等位基因以及TCR上的非HLA、非TCR基因 单阳性和双阴性细胞库，以及 确定这些基因可以在多大程度上改变基因库， 通过“积极”或“消极”的选择。这种对人TCR的影响 剧目可能有深远的影响，在确定一个 个体对病原体的反应能力和/或 自身免疫性疾病我们建议在一个高度近交的动物身上进行这些研究。 人口阿米什人，使用V节特异性单克隆抗体， qPCR以及其他正在开发的方法。我们特别 提议： 1)分析双阴性（DN）患者中的Valpha和Vbeta TCR库 同卵双胞胎和HLA相同和不相同的同胞中的细胞。这 这将有助于我们确定这种T细胞是否可以表达“禁止” TCR，可能是潜在的自身反应性。 2)确定HLA和非HLA基因对Valpha TCR的影响 单阳性（CD 4+和CD 8+）细胞库。 3)确定增加的数字关联的分子基础 表达Valpha2.3和HLA特异性B38的T细胞。 4)通过HLA基因鉴定“阳性”和“阴性”选择实例 在阿米什人中：首先，确定HLA关联的其他例子 然后，确定TCR库在多大程度上 被HLA等位基因改变了 5)确定在阿米什人中TCR由基因决定的例子， 非HLA、非TCR基因，其可代表内源性“超抗原”。 6)继续开发分析TCR的新方法 这将使它能够更容易地确定的影响， 单个HLA等位基因或TCR库上的内源性“超抗原”。

The broad goals of this proposal are to identify examples of the influence of individual HLA alleles as well as non-HLA, non-TCR genes on the TCR repertoire of both single-positive and double-negative cells, and to determine the extent to which these genes can alter the repertoire either by "positive" or "negative" selection. Such influences on the human TCR repertoire may have profound implications in terms of determining an individual's ability to respond to pathogens and/or development of an autoimmune disease. We propose to perform these studies on a highly inbred population. the Amish, using V-segment-specific monoclonal antibodies and qPCR as well as other methods that are being developed. Specifically, we propose to: 1) analyze the Valpha and Vbeta TCR repertoire in double-negative (DN) cells in identical twins and HLA-identical and non-identical sibs. This will help us in determining whether such T cells can express "forbidden" TCR which may be potentially autoreactive. 2) determine the influence of HLA and non-HLA genes on the Valpha TCR repertoire of single positive (CD4+ and CD8+) cells. 3) determine the molecular basis for the association of increased numbers of T cells expressing Valpha2.3 and the HLA specificity B38. 4) identify examples of "positive" and "negative" selection by HLA genes in the Amish: by-first, identifying additional examples of HLA associations with TCR and then, determining-the extent to which the TCR repertoire has been altered by the presence of that HLA allele. 5) identify examples in the Amish where TCR are genetically determined by non-HLA, nonTCR genes which may represent endogenous "superantigens". 6) continue the development of new methods for analyzing the TCR repertoire which will make it possible to more easily define the effects of individual HLA alleles or endogenous "superantigens" on the TCR repertoire.