Struyk L, Hawes G E, Dolhain R J, van Scherpenzeel A, Godthelp B, Breedveld F C, van den Elsen P J
Department of Immunohaematology, University Hospital Leiden, The Netherlands.
Int Immunol. 1994 Jun;6(6):897-907. doi: 10.1093/intimm/6.6.897.
In this study we have analyzed the TCR V alpha and V beta regions at the DNA level in the CD4+CD45RO+ memory T cell population of synovial tissue infiltrating T lymphocytes of three rheumatoid arthritis (RA) patients and one patient with chronic arthritis. Cell lines of CD4+CD45RO+, CD4+CD45RO-, CD8+CD45RO+ and CD8+CD45RO- T lymphocyte populations were generated following FACS cell sorting of freshly isolated synovial tissue mononuclear cell infiltrates (STMC) and of freshly isolated peripheral blood mononuclear cells (PBMC) of these patients. The phenotypic and molecular analyses have revealed the following. (i) The TCR repertoires of tissue infiltrating T lymphocytes in the various subsets were extensive on the basis of TCR V gene family usage. (ii) Furthermore, each patient displayed individual specific TCR V gene expression patterns in the various STMC and PBMC derived T cell subsets. However, the majority of these arthritis patients manifested increased expression of multiple TCR V gene families in the synovial tissue derived CD4+CD45RO+ T cell population when compared with the peripheral blood derived CD4+CD45RO+ subset. Of these gene families, we found enhanced expression of the TCR V alpha 7 and V beta 11 gene segments in synovial tissue to be shared by all four patients analyzed. (iii) Nucleotide sequence analysis of the CDR3 regions of a number of TCR V regions in the CD4+CD45RO+ T cell subsets has revealed that the CDR3 regions comprised within synovial tissue derived TCR V regions differed from those found in peripheral blood derived TCR V regions. These differences in CDR3 diversity might be the consequence of a specific interaction with particular MHC-peptide complexes expressed at the site of inflammation. (iv) The CDR3 region analysis also showed individual specific amino acid motifs within the N-D-N regions of all analyzed TCR V beta genes derived from PBMC as well as STMC.
在本研究中，我们在DNA水平分析了3例类风湿性关节炎（RA）患者和1例慢性关节炎患者滑膜组织浸润性T淋巴细胞的CD4+CD45RO+记忆T细胞群体中的TCR Vα和Vβ区域。在对这些患者新鲜分离的滑膜组织单核细胞浸润物（STMC）和新鲜分离的外周血单核细胞（PBMC）进行荧光激活细胞分选（FACS）后，生成了CD4+CD45RO+、CD4+CD45RO-、CD8+CD45RO+和CD8+CD45RO- T淋巴细胞群体的细胞系。表型和分子分析揭示了以下几点。（i）基于TCR V基因家族的使用情况，各个亚群中组织浸润性T淋巴细胞的TCR库广泛。（ii）此外，每位患者在各种源自STMC和PBMC的T细胞亚群中均表现出个体特异性的TCR V基因表达模式。然而，与外周血来源的CD4+CD45RO+亚群相比，这些关节炎患者中的大多数在滑膜组织来源的CD4+CD45RO+ T细胞群体中表现出多个TCR V基因家族的表达增加。在这些基因家族中，我们发现滑膜组织中TCR Vα7和Vβ11基因片段的表达增强为所有4例分析患者所共有。（iii）对CD4+CD45RO+ T细胞亚群中多个TCR V区域的CDR3区域进行核苷酸序列分析表明，滑膜组织来源的TCR V区域内包含的CDR3区域与外周血来源的TCR V区域中的不同。CDR3多样性的这些差异可能是与炎症部位表达的特定MHC-肽复合物特异性相互作用的结果。（iv）CDR3区域分析还显示，源自PBMC以及STMC的所有分析TCR Vβ基因的N-D-N区域内存在个体特异性氨基酸基序。
