Weiss A, Stobo J D
J Exp Med. 1984 Nov 1;160(5):1284-99. doi: 10.1084/jem.160.5.1284.
The association between T3 and the T cell antigen receptor was examined using the T3 bearing T cell leukemic line Jurkat. A monoclonal antibody, C305, was produced, which reacted with idiotypic-like determinants expressed on Jurkat. The molecule with which this antibody reacted was a disulfide-linked heterodimer of 90 kD, composed of polypeptides of 42 and 54 kD. Thus, C305 reacted with a molecule with characteristics of the putative T cell antigen receptor described by others. A series of mutants of Jurkat, induced with ethyl methane sulfonate or radiation, was selected for T3 or antigen receptor negativity. In every instance, there was a concomitant loss of both T3 and the antigen receptor as assessed by quantitative absorption, indirect immunofluorescence, and antibody plus complement-mediated cytotoxicity. The absence of antigen receptor molecules was confirmed on diagonal gels, excluding the possibility that conformational changes of the antigen receptor on such T3-negative mutants were responsible for the failure of such mutants to react with C305. Moreover, in a mutant that expressed a marked decrease in the level of T3 expression, there was a comparable decrease in the expression of antigen receptor determinants. These results suggest that there is an obligate requirement for the coexpression of T3 and the T cell antigen receptor. Furthermore, attempts to activate such mutants with the lectin phytohemagglutinin suggested that the expression of T3 and/or the antigen receptor was required for activation of these cells.
利用携带T3的T细胞白血病细胞系Jurkat研究了T3与T细胞抗原受体之间的关联。制备了一种单克隆抗体C305，它能与Jurkat细胞上表达的类独特型决定簇发生反应。与该抗体发生反应的分子是一个由42kD和54kD多肽组成的90kD二硫键连接的异二聚体。因此，C305与一种具有其他人所描述的假定T细胞抗原受体特征的分子发生反应。选择了一系列用甲磺酸乙酯或辐射诱导产生的Jurkat突变体，使其T3或抗原受体呈阴性。在每一个实例中，通过定量吸收、间接免疫荧光以及抗体加补体介导的细胞毒性评估，发现T3和抗原受体同时缺失。在对角线凝胶上证实了抗原受体分子的缺失，排除了此类T3阴性突变体上抗原受体的构象变化导致这些突变体无法与C305发生反应的可能性。此外，在一个T3表达水平显著降低的突变体中，抗原受体决定簇的表达也有类似程度的降低。这些结果表明，T3和T细胞抗原受体的共表达是必不可少的。此外，用凝集素植物血凝素激活此类突变体的尝试表明，这些细胞的激活需要T3和/或抗原受体的表达。
