McConnell T J, Yokoyama W M, Kikuchi G E, Einhorn G P, Stingl G, Shevach E M, Coligan J E
Biological Resources Branch, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
J Immunol. 1989 Apr 15;142(8):2924-31.
To assess the diversity of TCR delta-gene expression in dendritic epidermal T cells (DETC), we characterized the delta-gene used by a panel of cell lines that express the gamma delta TCR on the cell surface. Northern hybridization analyses with a panel of V delta probes representing each of the six known V delta families showed that each of these lines expressed either V delta 1 or V delta 6 containing transcripts. Southern hybridization analysis with the V delta probes gave rearrangement patterns consistent with Northern hybridization results. The correlation of V delta expression with V gamma expression revealed that not only may V delta usage be restricted in DETC cells, but that the pairing of gamma- and delta-chains may not be random. In the DETC cells examined, V delta 1 chains paired exclusively with V gamma 3C gamma 1 chains and V delta 6 chains paired with C gamma 2 and C gamma 4 chains. Sequence analysis of delta-cDNA clones corresponding to the expressed delta-chains from one of the V delta 1 expressing cell lines and two of the V delta 6 expressing cell lines revealed that, although the V delta 1 gene segment sequence and one of the V delta 6 gene segment sequences were identical to previously published V delta sequences, considerable variable region diversity was generated by complex V-D-J rearrangement patterns that utilized various combinations of N-additions, D delta 2 and possibly D delta 1 segments, and J delta 1 or J delta 2 segments. These complex rearrangement patterns distinguish these DETC lines from early thymocytes and suggest that, if DETC are thymic derived, they originate from relatively mature thymic cells.
为评估树突状表皮T细胞(DETC)中TCRδ基因表达的多样性,我们对一组在细胞表面表达γδTCR的细胞系所使用的δ基因进行了特征分析。用一组代表六个已知Vδ家族的Vδ探针进行Northern杂交分析,结果显示这些细胞系中的每一个都表达含有Vδ1或Vδ6的转录本。用Vδ探针进行Southern杂交分析,得到的重排模式与Northern杂交结果一致。Vδ表达与Vγ表达的相关性表明,不仅在DETC细胞中Vδ的使用可能受到限制,而且γ链和δ链的配对可能不是随机的。在所检测的DETC细胞中,Vδ1链仅与Vγ3Cγ1链配对,Vδ6链与Cγ2和Cγ4链配对。对来自一个表达Vδ1的细胞系和两个表达Vδ6的细胞系中表达的δ链对应的δ-cDNA克隆进行序列分析,结果显示,尽管Vδ1基因片段序列和其中一个Vδ6基因片段序列与先前发表的Vδ序列相同,但通过复杂的V-D-J重排模式产生了相当大的可变区多样性,这些重排模式利用了N-添加、Dδ2以及可能的Dδ1片段和Jδ1或Jδ2片段的各种组合。这些复杂的重排模式将这些DETC细胞系与早期胸腺细胞区分开来,并表明,如果DETC来源于胸腺,它们起源于相对成熟的胸腺细胞。