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Analysis of T cell antigen receptor (TCR) expression by human peripheral blood CD4-8- alpha/beta T cells demonstrates preferential use of several V beta genes and an invariant TCR alpha chain

机译:人类外周血CD4-8-α/βT细胞对T细胞抗原受体(TCR)表达的分析表明,优先使用了多个Vβ基因和不变的TCRα链

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摘要

CD4-CD8- (double negative [DN]) alpha/beta T cells are a largely uncharacterized subpopulation of unknown function. To investigate whether these cells are selected to recognize particular antigens or antigen-presenting molecules, DN alpha/beta T cells were purified from the peripheral blood of five normal donors and their T cell receptor (TCR) alpha and beta chains were examined. Random cloning of TCR alpha chains by single-sided polymerase chain reaction (PCR) amplification identified an invariant rearrangement between V alpha 24 and J alpha Q, with no N region diversity, which was expressed preferentially by DN alpha/beta T cells from all donors. Random cloning also identified a precise V alpha 7.2-J alpha (IGRJa14) rearrangement, with two variable amino acids encoded in the V-J junction, which was enriched in the DN alpha/beta T cell preparations from some, but not all, donors. Analysis of TCR beta chains by quantitative PCR amplification demonstrated that the expression of four V beta gene families, V beta 2, 8, 11, and 13, was markedly increased in these DN alpha/beta T cell preparations. The expression of particular TCRs by DN alpha/beta T cells from multiple donors indicates that these cells, or at least a subpopulation of cells with this phenotype, recognize a limited spectrum of antigens and suggests that they may use nonpolymorphic antigen-presenting molecules.
机译:CD4-CD8-(双阴性[DN])α/βT细胞是未知功能的很大程度上未知的亚群。为了研究是否选择了这些细胞来识别特定的抗原或抗原呈递分子,从五个正常供体的外周血中纯化了DNα/βT细胞,并检查了它们的T细胞受体(TCR)α和β链。通过单侧聚合酶链反应(PCR)扩增随机克隆TCRα链,发现V alpha 24和J alpha Q之间存在不变的重排,没有N区多样性,所有供体的DN alpha / beta T细胞优先表达。随机克隆还确定了精确的V alpha 7.2-J alpha(IGRJa14)重排,在V-J连接处编码了两个可变氨基酸,这些氨基酸在某些(但不是全部)供体的DN alpha / beta T细胞制剂中富集。通过定量PCR扩增对TCR beta链的分析表明,在这些DN alpha / beta T细胞制备物中,四个V beta基因家族(V beta 2、8、11和13)的表达显着增加。来自多个供体的DN alpha / beta T细胞表达特定的TCR,表明这些细胞或具有该表型的细胞的至少一个亚群可以识别有限范围的抗原,并暗示它们可以使用非多态性抗原呈递分子。

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