Bröker B M, Tsygankov A Y, Müller-Fleckenstein I, Guse A H, Chitaev N A, Biesinger B, Fleckenstein B, Emmrich F
Max-Planck-Gesellschaft, Arbeitsgruppen für Rheumatologie am Institut für Klinische Immunologie, Universität Erlangen-Nürnberg, Germany.
J Immunol. 1993 Aug 1;151(3):1184-92.
Investigation of human activated T cells has been complicated by the need for periodic restimulation with Ag/mitogen and accessory cells and by the limited life span of most human T cell clones. To overcome these problems, we have transformed established human T cell clones to permanent growth with Herpesvirus saimiri, a lymphoma-inducing virus of nonhuman primates. Three human CD4+ T cell clones were investigated in detail. They have been growing in the presence of exogenous IL-2 but without restimulation with mitogen or feeder cells for more than 11 mo with doubling times between 2 and 4 days. In contrast, their nontransformed parent clones needed to be restimulated with PHA and feeder cells every 14 to 21 days. To compare responses of H. saimiri-transformed clones with those of their parent clones, we stimulated the cells with IL-2 or with anti-CD3 and/or anti-CD4 mAb with and without cross-linking on the cell surface. Transformed and nontransformed T cell clones were strikingly similar in parameters of early signal transduction, namely, tyrosine phosphorylation and mobilization of calcium. Ligation of their TcR/CD3 complexes by mAb or by Ag in the presence of autologous accessory cells increased the proliferation and the secretion of IFN-gamma. Taken together, we have shown that human T cell clones immortalized with H. saimiri express functional CD3, CD4, and IL-2R. They constitute a simple, stable, reproducible and accessory cell-free model system for the investigation of signal transduction events in activated human T cells.
对人类活化T细胞的研究一直受到诸多因素的困扰，比如需要用抗原/丝裂原和辅助细胞进行定期再刺激，以及大多数人类T细胞克隆的寿命有限。为了克服这些问题，我们用赛氏疱疹病毒（一种非人类灵长类动物的淋巴瘤诱导病毒）将已建立的人类T细胞克隆转化为永生化生长。我们详细研究了三个人类CD4+ T细胞克隆。它们在外源性白细胞介素-2存在的情况下生长，无需丝裂原或饲养细胞的再刺激，已经生长了超过11个月，倍增时间在2到4天之间。相比之下，它们未转化的亲本克隆需要每14至21天用PHA和饲养细胞进行再刺激。为了比较赛氏疱疹病毒转化克隆与其亲本克隆的反应，我们用白细胞介素-2或抗CD3和/或抗CD4单克隆抗体在细胞表面交联或不交联的情况下刺激细胞。转化和未转化的T细胞克隆在早期信号转导参数方面，即酪氨酸磷酸化和钙动员方面，惊人地相似。在自体辅助细胞存在的情况下，用单克隆抗体或抗原连接它们的TcR/CD3复合物会增加增殖和干扰素-γ的分泌。综上所述，我们已经表明，用赛氏疱疹病毒永生化的人类T细胞克隆表达功能性的CD3、CD4和白细胞介素-2受体。它们构成了一个简单、稳定、可重复且无辅助细胞的模型系统，用于研究活化人类T细胞中的信号转导事件。
