The induction of HLA-DR antigen expression on U937 cells by interferon-gamma (IFN-gamma) is positively influenced by amount of fetal calf serum (FCS) added to the tissue culture medium. A transient alkalinization of FCS before its addition to the medium, dramatically decreased the immunomodulating activity of IFN-gamma. FCS was also found to be a dose-dependent enhancer of the IFN-gamma-induced 2',5'-oligoadenylate (2-5A) synthetase production. Our findings suggest the need for a serum factor(s), labile at basic pH values, to support at least two of the multiple IFN-gamma activities.