Struyk L, Hawes G E, Haanen J B, de Vries R R, van den Elsen P J
Department of Immunohematology and Blood Bank, University Hospital Leiden, The Netherlands.
Hum Immunol. 1995 Dec;44(4):220-7. doi: 10.1016/0198-8859(95)00110-7.
In order to establish whether specific MHC class II-peptide complexes are capable of selecting TCR V regions, we investigated in detail the TCR beta chain used in the recognition of HLA-DR3 restricted hsp65 peptide 3-13 in a tuberculoid leprosy patient. Using RT-PCR, a clear dominance of the TCRBV5 gene family was observed in a hsp65 peptide 3-13-specific T-cell line; however, not in fresh, unstimulated PBMCs, PHA-stimulated PBMCs, or a T-cell line specific for tetanus toxoid. DNA sequence analysis of the TCR V regions, comprising TCRBV5 genes, derived from the hsp65 peptide 3-13-specific T-cell line revealed the exclusive usage of the TCRBV55S1 gene segment and a predominance of one V-D-J gene rearrangement, which is indicative of clonal expansion of these T lymphocytes. Additional highly similar V-D-J gene rearrangements were detected at a low level in this hsp65 peptide 3-13 specific T-cell line. These conserved junctional regions (CDR3 regions) could not be detected within the TCRBV5 gene family of fresh PBMCs, PHA-stimulated PBMCs, hsp65, and tetanus-toxoid-specific T-cell lines from this patient. The observations in this tuberculoid leprosy patient reveal that an HLA class-II-restricted T-cell response results in selection of TCRBV regions which are highly similar in amino acid composition to the CDR3 region within the expanding TCRBV regions.
为了确定特定的MHC II类肽复合物是否能够选择TCR V区,我们详细研究了一名结核样麻风病患者识别HLA - DR3限制性hsp65肽3 - 13时所使用的TCRβ链。利用逆转录聚合酶链反应(RT-PCR),在hsp65肽3 - 13特异性T细胞系中观察到TCRBV5基因家族明显占优势;然而,在新鲜的未刺激外周血单个核细胞(PBMC)、PHA刺激的PBMC或破伤风类毒素特异性T细胞系中并非如此。对来自hsp65肽3 - 13特异性T细胞系的包含TCRBV5基因的TCR V区进行DNA序列分析,结果显示TCRBV55S1基因片段被唯一使用,且一种V-D-J基因重排占主导,这表明这些T淋巴细胞发生了克隆性扩增。在该hsp65肽3 - 13特异性T细胞系中还低水平检测到了其他高度相似的V-D-J基因重排。在该患者的新鲜PBMC、PHA刺激的PBMC、hsp65和破伤风类毒素特异性T细胞系的TCRBV5基因家族中未检测到这些保守的连接区(CDR3区)。该结核样麻风病患者的观察结果表明,HLA II类限制性T细胞反应导致选择了TCRBV区,这些区域的氨基酸组成与扩增的TCRBV区内的CDR3区高度相似。