Kraig E, Kronenberg M, Kapp J A, Pierce C W, Abruzzini A F, Sorensen C M, Samelson L E, Schwartz R H, Hood L E
J Exp Med. 1983 Jul 1;158(1):192-209. doi: 10.1084/jem.158.1.192.
We have attempted to determine whether T cells and B cells that have the same antigenic specificity and whose receptors share idiotypic determinants in fact express similar VH gene segments. To do this, we have obtained and characterized a cDNA clone containing the entire coding sequence for the VH gene from a glutamic acid60/alanine30/tyrosine10 (GAT)-binding immunoglobulin that carries the CGAT idiotype. The GAT-VH clone was hybridized to Northern blots of GAT-specific T cell RNAs; there was no evidence of a T cell transcript that hybridized to the GAT-VH probe. The T cells analyzed included: (a) 10 GAT-binding suppressor T cell hybridomas, 6 of which secreted factors with CGAT idiotypic determinants, (b) one GAT-specific helper T cell hybridoma, and (c) two GAT-specific helper T cell lines grown in the absence of feeder cells. The detection limit of the Northern blot analysis was 1-2 copies of a particular mRNA species per cell for the hybridomas and 5-10 copies per cell for the T cell lines. Therefore, we conclude that T and B lymphocytes responding to GAT do not utilize similar VH gene segments. Furthermore, the presence of idiotypic determinants on T lymphocytes does not necessarily imply close structural similarity between T and B cell antigen receptors.
我们试图确定具有相同抗原特异性且其受体共享独特型决定簇的T细胞和B细胞实际上是否表达相似的VH基因片段。为此,我们获得并鉴定了一个cDNA克隆,该克隆包含来自携带CGAT独特型的谷氨酸60/丙氨酸30/酪氨酸10(GAT)结合免疫球蛋白的VH基因的完整编码序列。将GAT-VH克隆与GAT特异性T细胞RNA的Northern印迹杂交;没有证据表明存在与GAT-VH探针杂交的T细胞转录本。分析的T细胞包括:(a)10个GAT结合抑制性T细胞杂交瘤,其中6个分泌具有CGAT独特型决定簇的因子,(b)1个GAT特异性辅助性T细胞杂交瘤,以及(c)在无饲养细胞的情况下培养的2个GAT特异性辅助性T细胞系。Northern印迹分析的检测限对于杂交瘤为每个细胞1 - 2个特定mRNA种类的拷贝,对于T细胞系为每个细胞5 - 10个拷贝。因此,我们得出结论,对GAT作出反应的T淋巴细胞和B淋巴细胞不利用相似的VH基因片段。此外,T淋巴细胞上独特型决定簇的存在并不一定意味着T细胞和B细胞抗原受体之间存在密切的结构相似性。