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JCS ePress
online publication date 18 Jan 2005
doi: 10.1242/jcs.01640
Research Article
Induction of apoptosis by TNF receptor 2 in a T-cell hybridoma is FADD dependent and blocked by caspase-8 inhibitors
Bart Depuydt,
Geert Van Loo,
Peter Vandenabeele*,
and
Wim Declercq
* Author for correspondence (e-mail: peter.vandenabeele{at}dmbr.ugent.be)
Previously we reported that both human TNFR1 and TNFR2 mediate TNF-induced apoptosis in the transfected rat/mouse T cell hybridoma PC60. We show here that TNFR2-mediated apoptosis in PVC60 cells can be blocked by the broad-spectrum caspase inhibitor zVAD-fmk, the caspase-8 inhibitor zIETD-fmk and by CrmA, a viral inhibitor of caspase-1 and caspase-8. This suggests an involvement of caspase-8 in TNFR2-mediated apoptosis. The upstream adaptor of caspase-8, FADD, is also involved in TNFR2-induced cell death, since transient overexpression of a dominant negative deletion mutant of FADD inhibited apoptosis induced by this receptor. TNFR2-induced apoptosis is independent of endogenous TNF or other death-inducing ligand production and subsequent activation of TNFR1 or other death receptors. Furthermore, TNFR2 stimulation does not enhance sensitivity for a subsequent TNFR1-induced apoptotic signal, as has been reported for Jurkat cells. TRAF2 downregulation, which has been proposed as the mechanism by which TNFR2 enhances TNFR1 signaling, was observed in PC60 cells, but the TNRF1 signal was not modulated. These data confirm the capacity of TNFR2 to generate an apoptotic cell death signal independent of TNFR1.
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