Regulation of cell survival in CD95-induced T cell apoptosis: role of NF-kappa B/Rel transcription factors

The activity of NF-kappa B/Rel transcription factors can inhibit the apopto sis induced by TNF, UV or cancer therapy drugs in a number of cell types, i ncluding human T lymphocytes. Furthermore, the NF-kappa B/Rel inducer, phor bol-12-myristate-13-acetate (PMA), has been reported to suppress the CD95-i nduced apoptosis of human T lymphocytes. To verify whether the survival-enh ancing effect of PMA required NF-kappa B/Rel activity, we generated two Jur kat cell sublines (AL.7 and AL.8) transfected with a pCMV4-I kappa B alpha construct, and two (AL.3 and AL.5) with the void pCMV4 vector. Compared to wild type, AL.3 and AL.5 cells, the AL.7 and AL.8 sublines displayed marked ly lower amounts of NF-kappa B/Rel nuclear complexes and a reduced expressi on of a kappa B-controlled CAT reporter gene after 1 and 4 h of incubation with PMA, respectively. All the five cell types displayed negligible levels of apoptosis when cultured with medium or PMA alone; when stimulated with the mAb CH-11, the AL.7 and AL.8 sublines displayed apoptotic responses onl y slightly (< 0.5 fold) higher than control cells. On the other hand, the s alvage activity of PMA was partially impaired in the AL.7 and AL.8 sublines . PMA inhibited apoptosis by > 85% in wild type, AL.3 and AL.5 cells and by < 60% in the AL.7 and AL.8 sublines; the apoptosis percentages in the mAb CH-11 + PMA cultures of the I kappa B alpha-transfected cells were > 4-fold higher than in control cells. We conclude that the inhibition of the CD95- induced apoptosis by PMA relies on both NF-kappa B/Rel-dependent and -indep endent mechanisms. The partial contribution of these nuclear factors to the suppression of apoptosis indicates that the NF-kappa B/Rel activity can in fluence the extent of the CD95-induced T cell death.