T-CELL RECEPTOR-INDUCED FAS LIGAND EXPRESSION IN CYTOTOXIC T-LYMPHOCYTE CLONES IS BLOCKED BY PROTEIN-TYROSINE KINASE INHIBITORS AND CYCLOSPORINE-A
T-CELL RECEPTOR-INDUCED FAS LIGAND EXPRESSION IN CYTOTOXIC T-LYMPHOCYTE CLONES IS BLOCKED BY PROTEIN-TYROSINE KINASE INHIBITORS AND CYCLOSPORINE-A
复制标题
DOI：
10.1002/eji.1830241032
复制
发表时间：
1994-10-01
影响因子：
5.4
通讯作者：
GOLSTEIN, P
中科院分区：
文献类型：
--
作者：
ANEL, A;BUFERNE, M;GOLSTEIN, P
Fas/APO-1是肿瘤坏死因子受体蛋白家族的成员，当与单克隆抗体（mAb）或其生理配体交联时，其诱导凋亡。最近，已经提出了基于穿孔素和基于Fas的机制来解释T细胞介导的细胞毒性。在本研究中，我们使用了对H-2 K（B）具有特异性的鼠CD 8（+）细胞毒性T淋巴细胞（CTL）克隆（KB5.C20）和同一克隆的T细胞受体（TcR）阴性变体（2005（-）D4），以检测（i）同一细胞是否可以发挥两种细胞毒性效应机制和（ii）TcR参与诱导基于Fas的细胞毒性的作用。我们证明了TcR（+）和TcR（-）克隆在佛波酯/离子霉素刺激后都能表达Fas配体，并且通过携带抗原的细胞或其抗克隆型单克隆抗体，（Desire-1）也能够诱导基于Fas的细胞毒性，其导致Ca 2+依赖性的、推测为基于穿孔素的细胞毒性。此外，使用抑制剂，我们研究了参与诱导基于Fas的细胞毒性和在CTL克隆中Fas配体mRNA表达的信号转导途径。src样蛋白酪氨酸激酶（PTK）抑制剂除莠霉素A的抑制作用强烈表明，src样蛋白酪氨酸激酶（PTK）通过TcR接合参与Fas配体诱导。抑制Fas配体诱导的染料木黄酮，一个更一般的TPK抑制剂，即使在刺激PMA加离子霉素，建议可能参与的PTK活动下游的蛋白激酶C（PKC）在Fas配体诱导的CTL。最后，通过用环孢菌素A部分抑制Fas配体诱导，证实了Ca 2 +/钙调素依赖性蛋白磷酸酶钙调神经磷酸酶在Fas配体诱导中的意义。因此，在CTL克隆中，Fas配体的表达是可诱导的TcR参与通过一个途径类似的参与一些淋巴因子基因的表达。
Fas/APO-1 is a member of the tumor necrosis factor receptor family of proteins, that induces apoptosis when cross-linked with monoclonal antibody (mAb) or with its physiological ligand. Recently, both a perforin-based and a Fas-based mechanism have been proposed to account for T cell-mediated cytotoxicity. In the present study we used a murine CD8(+) cytotoxic T lymphocyte (CTL) clone (KB5.C20) specific for H-2K(b) and a T cell receptor (TcR)-negative variant of the same clone (2005(-)D4) to test (i) whether the same cell can exert both cytotoxic effector mechanisms and (ii) the role of TcR engagement in the induction of Fas-based cytotoxicity. We demonstrate that both the TcR(+) and TcR(-) clones were able to express the Fas ligand after stimulation with phorbol 12-myristate 13-acetate (PMA)/ionomycin, and that TcR engagement of the KB5.C20 clone by means of antigen-bearing cells or of its anticlonotypic mAb (Desire-1), which leads to Ca2+-dependent, presumably perforin-based, cytotoxicity, was also able to induce Fas-based cytotoxicity. In addition, using inhibitors we investigated the signal transduction pathway(s) involved in the induction of Fas-based cytotoxicity and expression of the Fas ligand mRNA in the CTL clones. The involvement of src-like protein tyrosine kinases (PTK) in Fas ligand induction through TcR engagement, was strongly suggested by inhibition with the src-like PTK inhibitor herbimycin A. Inhibition of Fas ligand induction by genistein, a more general TPK inhibitor, even upon stimulation by PMA plus ionomycin, suggested the possible involvement of PTK activities downstream of protein kinase C (PKC) in Fas ligand induction in CTL. Finally, the implication of the Ca2+/calmodulin-dependent protein phosphatase calcineurin in Fas ligand induction was demonstrated by the partial inhibition of Fas ligand induction with cyclosporin A. Thus, in CTL clones, Fas ligand expression is inducible by TcR engagement through a pathway similar to that involved in expression of some lymphokine genes.