We have investigated the effect of cytokines, including interleukin-6 (Il-6), interleukin-la (Il-la), and tumor necrosis factor-alpha ("Fa), on the inducible expression of cytochrome P450s (CYP) CYPlA1, CYPlA2, and CYP3A4 in human hepatocytes in primary culture. The ability of these cultures to mimic the acute phase response when stimulated with cytokines was evaluated using immunoblotting to measure the production of albumin, ferritin, fibrinogen, and ceruloplasmin. The cytokines exhibited specific patterns of action on the production of these proteins. Albumin was depressed by all the cytokines. In contrast to 11-6 and Il-la, T " -a reduced the production of fibrinogen and ceruloplasmin but stimulated the production of ferritin. When cells were treated with the CYP inducer alone, large increases in the expression of CYPlAl and CYPlA2 by P-naphthoflavone and of CYP3A4 by rifampicin were observed at messenger RNA (mRNA) and protein levels, by ribonuclease protection and immunoblotting, respectively. When the cells were treated with the inducer plus cytokines, the induction of mRNA was greatly reduced. Again, specific patterns of action were revealed: 11-6 had the most potent effect on CYP3A4, whereas TNF-a was the most potent with CYPlA genes. In all cases, changes at the protein levels paralleled changes at the mRNA levels. In cells preinduced with P-naphthoflavone or rifampicin, the decay with time of the levels of the CYPlA2 or CYP3A4 proteins, after the removal of the inducer, was not affected by cytokines. We conclude that cytokines strongly repress the inducibility of CYPlAs and CYP3A4 genes at a transcriptional or a posttranscriptional level, but affect neither the rate of translation of CYP mFWAs nor the rate of degradation of the CYP proteins in these cultures. (HEPATOLOGY 1995; 221 143-1 153.) Abbreviations: CYP, cytochrome P450s; TNF-a, tumor necrosis factoralpha; 11-10, interleukin-1 alpha; mRNA, messenger RNA, cDNA, complementary DNA, NO, nitric oxide.From the 'INSERM U 128,