We evaluated the effect of IFN-beta on the expression of transferrin receptor (TfR) during the in vitro differentiation of peripheral blood monocytes to macrophages. IFN-beta exerted a strong inhibitory effect on the expression of TfR. As little as 0.1 IU/ml was sufficient to induce a 40% reduction of transferrin (Tf) binding sites on 7-day cultured macrophages. Scatchard plot analysis revealed that this impaired Tf binding in IFN-beta-treated macrophages was not due to a decreased affinity of the TfR for its ligand but to a reduction in the number of cell surface TfR. IFN-gamma did not exert any significant effect on the expression of TfR, even though it was capable of partially reverting the inhibitory effect of the IFN-beta on Tf binding. To understand the mechanism by which IFN-beta inhibited TfR expression, we examined the expression of TfR mRNA, 125I-Tf binding to detergent-solubilized cells, and TfR cellular distribution. The results of these experiments showed that IFN-beta caused neither a significant alteration of the expression of TfR mRNA nor a decrease of the total content of TfR molecules. Moreover, immunofluorescence analysis of TfR localization indicated that TfR was clustered in an intracellular compartment in IFN-beta-treated macrophages. These data demonstrate that IFN-beta is capable of dramatically down-modulating TfR in macrophages by post-translational mechanisms (i.e., by sequestering this receptor in intracellular compartments).