The relationship of Ia expression and antigen-presenting function by macrophages has been evaluated. When macrophages are maintained in standard culture media, both Ia antigens and accessory cell function are lost. The reacquisition of these properties follows exposure to an Ia-inducing lymphokine, for which cDNA-derived interferon-gamma may substitute. The induction of function is related quantitatively to the level of Ia expression. Moreover, both properties reflect newly expressed Ia determinants, since treatment with anti-I-A plus complement at the beginning of culture diminishes neither the subsequent level of Ia expression nor function. Treatment with anti-Mac-1 plus complement, however, reduces function commensurate with the effectiveness of macrophage depletion. Finally, we find that fixation of macrophages after exposure to antigen does not inhibit antigen presentation, indicating that metabolic activity, while required for antigen processing, is not necessary for presentation.