Castelli C, Mazzocchi A, Salvi S, Anichini A, Sensi M
Division of Experimental Oncology D, Istituto Nazionale Tumori, Milan, Italy.
Scand J Immunol. 1992 Apr;35(4):487-94. doi: 10.1111/j.1365-3083.1992.tb02883.x.
We report here the molecular characterization of the T-cell receptor (TCR) expressed by a human HLA-A2 specific cytotoxic T-cell clone named CTL 49. Flow cytometry analysis with a panel of anti-TCR antibodies revealed an OKT3+, WT31+, A13+, BB3-, TCR delta-, delta TCS1-, TCR gamma/delta 1-, OKT4-, and OKT8+ phenotype, suggesting that, in CTL 49, the V delta 1-encoded A13 epitope could be included in its alpha beta TCR. Northern blot analysis confirmed the presence of C alpha, C beta and V delta 1 specific transcripts while no hybridization signal was detected by a C delta specific probe. Polymerase chain reaction (PCR) amplification of the first strand cDNA from CTL 49 with TCR-specific primers and sequence analysis revealed that V delta 1 region is productively rearranged to J alpha and to C alpha regions. This alpha chain pairs with a beta chain composed of V beta 13.2/D beta/J beta 2.3/C beta 2 leading to the expression of a functional TCR complex. These results, in addition to providing further evidence for the sharing of V delta 1 by alpha/beta and gamma/delta TCR, indicate that an alpha/beta T-cell receptor which includes the V delta 1 variable region can be involved in alloreactive recognition.
我们在此报告了一个名为CTL 49的人HLA - A2特异性细胞毒性T细胞克隆所表达的T细胞受体(TCR)的分子特征。用一组抗TCR抗体进行的流式细胞术分析显示其表型为OKT3 +、WT31 +、A13 +、BB3 -、TCR δ -、δTCS1 -、TCR γ/δ 1 -、OKT4 -和OKT8 +,这表明在CTL 49中,Vδ1编码的A13表位可能包含在其αβTCR中。Northern印迹分析证实了存在Cα、Cβ和Vδ1特异性转录本,而用Cδ特异性探针未检测到杂交信号。用TCR特异性引物对CTL 49的第一链cDNA进行聚合酶链反应(PCR)扩增并进行序列分析,结果显示Vδ1区域有效地重排至Jα和Cα区域。该α链与由Vβ13.2 / Dβ / Jβ2.3 / Cβ2组成的β链配对,从而导致功能性TCR复合物的表达。这些结果,除了为α/β和γ/δTCR共享Vδ1提供了进一步的证据外,还表明包含Vδ1可变区的α/βT细胞受体可能参与同种异体反应性识别。