Addition of antibodies to major histocompatibility complex class I (MHC class I) and F(c) gamma RIII (CD16) antigens resulted in the synergistic augmentation of natural killer (NK) cell death, and the loss of NK cell cytotoxic function. The binding of anti-CD16 and anti-class I antibodies to the same population of NK cells is required for the synergistic augmentation of NK cell death. Moreover, the addition of antibodies to leukocyte function antigen-1 (LFA-1), which significantly inhibited the phorbol 12-myristate 13-acetate (PMA) and ionomycin mediated NK cell death, had no effect on NK cell death mediated by anti-CD16 and anti-class I antibodies. The increase in NK cell death was associated with an increase in tumor necrosis factor-alpha (TNF-alpha) secretion, and concomitant inhibition of nuclear factor kappa B (NFkappaB) activation and the induction of c-jun N-terminal kinase (JNK) activity in NK cells treated with the combination of anti-class I and anti-CD16 antibodies. Furthermore, the inhibition of NFkappaB activation in anti-CD16 and anti-class I antibody treated NK cells was paralleled with a significant increase in inhibitor of kappa B (IkappaB) protein expression. Overexpression of IkappaB super-repressor in YT, a NK cell line, caused significant up-regulation of TNF-alpha, PMA and ionomycin and Fusobacterium nucleatum mediated NK cell death. Overall, our studies suggest an important regulatory role for NFkappaB and JNK activities in MHC class I mediated NK cell death.