Qualitative and quantitative differences in the cellular responses mediated through Fas antigen and tumor necrosis factor receptor.
Like tumor necrosis factor (TNF), antibodies against the Fas antigen (anti-Fas) are cytotoxic to some and induce proliferation of other Fas-expressing cells. In this study, we compared cellular responses mediated through TNF with anti-Fas using a T cell line (Jurkat) and a macrophage cell line (U-937). These two cell types differed in that the Jurkat cells expressed higher levels of Fas antigen than U-937 cells, whereas the latter expressed higher levels of the p80 form of the TNF receptor than Jurkat cells. Treatment for 72 h with anti-Fas inhibited the growth of both Jurkat and U-937 cells, the 50% inhibitory concentrations (IC50) being 10 and 100 ng/ml, respectively. Under similar conditions, the IC50 for TNF was > 100 and 0.8 ng/ml for Jurkat and U-937 cells, respectively. Like TNF, the cytotoxic effects of anti-Fas were potentiated by cycloheximide, showing they did not require protein synthesis. Interestingly, in the presence of cycloheximide, the kinetics of cell killing was more rapid for TNF than anti-Fas (50% inhibition occurred at 3 versus 6h). Treatment of both cell types with anti-Fas led to time-dependent DNA fragmentation, but TNF-induced DNA fragmentation occurred only in the presence of cycloheximide. Pretreatment of cells with TNF led to resistance to TNF but not to anti-Fas, suggesting that the receptors for the two are not cross-modulated. Furthermore, TNF activated the nuclear transcriptional factor NF-kappa B in both cell types, whereas anti-Fas had no effect. Overall, our results demonstrate that anti-Fas and TNF transduce over-lapping and nonoverlapping signals in macrophage-like and T cell lines through distinct pathways.
Totpal
K
K
Department of Molecular Oncology, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Singh
S
S
Lapushin
R
R
Aggarwal
B B
BB
D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects / Q000276:immunology; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival / Q000276:immunology; D003513:Cycloheximide / Q000494:pharmacology; D003602:Cytotoxicity, Immunologic; D004249:DNA Damage; D008264:Macrophages / Q000166:cytology / Q000276:immunology*; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000096:biosynthesis; D018124:Receptors, Tumor Necrosis Factor / Q000502:physiology*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D019014:fas Receptor / Q000276:immunology*
D016328:NF-kappa B; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha; D019014:fas Receptor; D003513:Cycloheximide