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Regulation of TNF-Related Apoptosis-Inducing Ligand-Mediated Death-Signal Pathway in Human β Cells by Fas-Associated Death Domain and Nuclear Factor κB

2005; Elsevier BV; Volume: 66; Issue: 7 Linguagem: Inglês

10.1016/j.humimm.2005.03.009

1879-1166

Di Ou, X. Wang, Daniel L. Metzger, Marjorie Robbins, Junqi Huang, Christian Jobin, J. K. Chantler, Roger F.L. James, Paolo Pozzilli, Aubrey J. Tingle,

Immune Response and Inflammation

Transfectants of human CM and NES2Y β cell lines and primary islets transfected by FADD-DN (dominant-negative form of Fas-associated death domain), a mutant of FADD and/or a superrepressor of nuclear factor κB (NF-κB) (AdIκB(SA)2), were examined for their susceptibility to the TRAIL (TNF-related apoptosis-inducing ligand)-induced death signal pathway, compared with controls, wild-type cells, and vector transfectants in caspase fluorescence, Western blot, electrophoretic mobility shift, apoptosis, and cytotoxicity assays. FADD-DN inhibited caspase-8 activation induced by TRAIL in the transfectants of CM and NES2Y cells. TRAIL-induced apoptosis and cytotoxicity to the FADD-DN transfectants were decreased in comparison to those responses in controls (CM, p < 0.01 and p < 0.01; NES2Y, p < 0.05, and p < 0.02, respectively). When CM, NES2Y, and primary islet cells were transfected by AdIκB(SA)2, TRAIL-induced IκB degradation and nuclear translocation of NF-κB p50/p65 were blocked. TRAIL-induced apoptosis and cytotoxicity to AdIκB(SA)2transfectants of these cells were also reduced (CM, p < 0.02 and p < 0.02; NES2Y, p < 0.01 and p < 0.01, respectively, and islet p < 0.01 for cytotoxicity). Finally, cytotoxicity induced by TRAIL in CM and NES2Y cells transfected with both FADD-DN and AdIκB(SA)2 was reduced, compared with that observed in these cells transfected with either FADD-DN alone or AdIκB(SA)2 alone, suggesting that FADD and NF-κB have synergistic proapoptotic regulatory effects on the susceptibility of β cell lines and islet cells to TRAIL-induced destruction.