Both IFN-beta and TGF-beta have demonstrated their ability to antagonize several of the stimulatory activities of IFN-gamma on human macrophages, thereby classifying them as Th2-like. Aiming at a further characterization of their role in Th1/Th2 development, we studied their possible interaction with IL-12, the key Th1 cytokine. We found that IFN-beta by itself induced modest amounts of IFN-gamma, but was able to synergize with IL-12 for IFN-gamma induction. TGF-beta, on the other hand, had no effect by itself and inhibited significantly the IL-12-induced IFN-gamma secretion. The differential effect of IFN-beta and TGF-b on IL-12 bioactivity was most pronounced upon IFN-gamma synthesis, since IFN-beta induced only marginal amounts of IL-10 and IL-12 and TGF-beta diminished constitutive IL-10 production, while neither had a significant effect on TNF-alpha production. Although monocytes did not produce detectable IFN-gamma with any of the stimuli, adherent cells were found to cooperate with non-adherent lymphocytes for maximal IFN-gamma production. However, IL-18, a monocyte-derived IFN-gamma-inducing cytokine able to synergize with IL-12, was undetectable in IFN-beta or IFN-beta+IL-12-stimulated cells. In conclusion, the ability of IFN-beta to synergize with IL-12 for IFN-gamma synthesis, without significant concomitant IL-10 production, suggest a strong boost to Th1 development, which seems to be IL-18-independent.