| Abstrakt: |
Murine CD4+Th1 clones require de novosynthesis of proteins to express a cytotoxicity that is mediated by de novosynthesized Fas ligand (FasL). The cytotoxic process of the CD4+Th1 effectors can be separated into four stages, namely conjugate formation, activation, lethal hit, and effector-independent target cell death. The present study describes the cytotoxic process in terms of FasL induction and Fas/FasL molecular interactions for death signal transduction. Fas-Ig fusion proteins, cycloheximide, actinomycin D, and EGTA + MgCl2were used to analyze each stage of the cytotoxic process in terms of FasL/Fas participation. The results demonstrate that the activation-induced de novomRNA and protein synthesis were for FasL, which provided the predominant cytotoxic activity of CD4+Th1 effectors. Once activated, Th1 effectors express cytotoxic activity in the presence of EGTA + MgCl2an experimental [Ca2+]ext-independent condition characteristic of FasL-mediated cytotoxicity. The ability of Fas-Ig to inhibit target lysis declined rapidly after conjugate formation, indicating that FasL-mediated lethal hit is critically dependent on conjugate formation and, once delivered, the effector-independent target lysis proceeds. After the lethal hit stage, transduction of Fas-mediated death signal was independent of de novosynthesis of macromolecules in targets because treatments that inhibited more than 98% of the macromolecule synthesis had little effect on target lysis. Our study provides the first molecular view in terms of FasL/Fas of the cytotoxic process of CD+Th1 cells. |
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