Human vascular endothelial cells normally do not express class II MHC molecules in culture. IFN-gamma has been shown to induce expression of class I and class II MHC molecules on endothelial cell cultures from umbilical cord. We could detect these Ag by FACS analysis when endothelial cells were cultured for 3 days in the presence of 200 to 1000 U/ml of rIFN-gamma. Among the class II MHC molecules, HLA-DR and -DP but not -DQ were consistently induced. Addition of rIFN-alpha-D/A to IFN-gamma-treated cells inhibited the expression of class II MHC but not class I MHC molecules. Furthermore, the inhibition was more pronounced when IFN-alpha-D/A was added before or simultaneously as IFN-gamma. Natural IFN-alpha also exhibited similar inhibition and its suppressive effect was abolished in the presence of anti-IFN-alpha antibody. On the contrary, dexamethasone, a known inhibitor of class II MHC molecules on murine macrophages, showed a slight enhancing effect on class II MHC Ag. These results suggest an immunoregulatory role for IFN-alpha on non-lymphoid cells and that controlling elements for expression of class II MHC molecules may be different on various cell types as well as species.