Fas-mediated apoptosis is one of the immune effector pathways leading to the elimination of virus infected cells. In vivo, apoptotic signals are delivered to virus infected cells by Fas-L and other cytokines secreted by specific T lymphocytes. Cellular immune response appears to be essential in prevention of human cytomegalovirus (HCMV) disease. We have hypothesized that HCMV infection might directly or indirectly result in upregulation of Fas receptor and in the presence of Fas ligand, lead to apoptosis of infected cells. We show that infection of human fibroblasts with HCMV is associated with upmodulation of Fas-R process that could be further potentiated by interferon (IFN-gamma). Using DNA agarose gel electrophoresis, terminal dideoxy transferase reaction, and annexin assay, we demonstrated that in a productive HCMV infection of human fibroblasts, loss of cell viability was not only due to virus-mediated cell lysis but also to due to apoptosis. IFN-gamma induced relative HCMV resistance and prevented loss in cell viability. In contrast, anti-Fas monoclonal antibody CH11, serving as Fas agonist, resulted in an accelerated loss in viability of infected cells. IFN-gamma in combination with CH11 further increased the rate of apoptosis and compared to cultures with CH11 only, this effect was not restricted to only infected cells. While IFN-gamma did not affect the number of cells expressing immediate early antigen, it markedly reduced structural protein expression. IFN-gamma in combination with CH11, decreased the expression of HCMV matrix protein pp65, reduced the amount of HCMV DNA and infectious virus produced. Our results are consistent with the theory that cells infected with HCMV can be eliminated by immune effector cells via Fas-mediated apoptosis. IFN-gamma, in addition to its intrinsic antiviral activity, primes HCMV infected cells to the action of Fas ligand and Fas-mediated apoptosis.