Shirwan H, Chi D, Makowka L, Cramer D V
Department of Surgery, Cedars-Sinai Research Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048.
J Immunol. 1993 Nov 15;151(10):5228-38.
The T cell response to many self-MHC-restricted nominal Ag involves limited use of the TCR repertoire. The status of the TCR repertoire in allogeneic responses remains unclear. In this report, we have studied the TCR V beta gene repertoire involved in the rejection of cardiac allografts disparate for major and minor histocompatibility Ag in rats. Graft-infiltrating lymphocytes (GIL) were isolated from rejecting heart allografts and analyzed for the expression of the V beta repertoire using a cDNA library and a semiquantitative polymerase chain reaction (PCR). We report here that GIL isolated at early stages of the rejection reaction preferentially use the V beta 4 gene. First, V beta 4 comprised 36.4% (8/22) of randomly sequenced cDNA clones isolated from a TCR-beta chain-specific cDNA library established from GIL harvested 3 days posttransplantation. The V beta 4 gene in these clones was found in conjunction with several different J beta and N regions, suggesting a dominant role for the V beta 4 encoded domains in the recognition of allograft Ag. Second, the V beta 4 message comprised 56.6 to 65.7% of the transcripts expressed by the 20 rat V beta genes in three T cell lines established from GIL isolated 2 days posttransplantation. Third, fresh, unmanipulated GIL harvested at days 2 and 3 posttransplantation predominantly expressed the V beta 4 gene. Fourth, the expression of V beta 4 in naive splenocytes constituted only 5.4% of the V beta detected, suggesting that the predominant use of the V beta 4 gene by GIL was not a consequence of its high level of expression in the periphery. The limited use of the TCR repertoire in allograft rejection may provide the opportunity to interrupt the rejection process and induce donor-specific tolerance by targeting a select population of T cells for inactivation or elimination.
T细胞对许多自身MHC限制的名义抗原的反应涉及TCR库的有限使用。同种异体反应中TCR库的状态仍不清楚。在本报告中，我们研究了参与大鼠主要和次要组织相容性抗原不同的心脏同种异体移植排斥反应的TCR Vβ基因库。从排斥的心脏同种异体移植中分离出移植物浸润淋巴细胞（GIL），并使用cDNA文库和半定量聚合酶链反应（PCR）分析Vβ库的表达。我们在此报告，在排斥反应早期分离的GIL优先使用Vβ4基因。首先，Vβ4占从移植后3天收获的GIL建立的TCR-β链特异性cDNA文库中随机测序的cDNA克隆的36.4%（8/22）。在这些克隆中发现Vβ4基因与几个不同的Jβ和N区域结合，表明Vβ4编码结构域在同种异体移植抗原识别中起主导作用。其次，在从移植后2天分离的GIL建立的三个T细胞系中，Vβ4信息占20个大鼠Vβ基因表达的转录本的56.6%至65.7%。第三，移植后第2天和第3天收获的新鲜、未处理的GIL主要表达Vβ4基因。第四，幼稚脾细胞中Vβ4的表达仅占检测到的Vβ的5.4%，这表明GIL对Vβ4基因的优先使用不是其在外周高水平表达的结果。同种异体移植排斥反应中TCR库的有限使用可能提供机会，通过靶向选择的T细胞群体使其失活或消除来中断排斥过程并诱导供体特异性耐受。
