Cytomegaloviruses (CMVs) cause common viral infectious diseases and are difficult for the host immune system to eliminate, which leads to persistent or chronic infection. To investigate the T cell immune response stimulated by murine cytomegalovirus (MCMV) infection and the role of CD4(+)CD25(+)Foxp3(+) T regulatory cells (Tregs) in this process, T cells containing various proportions of Tregs were co-cultured with MCMV-infected mouse embryo fibroblasts (MEFs). MCMV infection stimulated proliferation of effector T cells as well as differentiation to Tregs, which consequently increased the expression of TGF-beta and IL-10. The proliferation of Tc1 (CD3(+)CD8(+)IFN-gamma(+)), Th1 (CD3(+)CD4(+)IFN-gamma(+)), and Tc2 (CD3(+)CD8(+)IL-4(+)) subsets was significantly suppressed with an increased proportion of Tregs in the co-culture system. Treg-depleted T cells inhibited viral load when co-cultured with MCMV-infected MEFs, however, this inhibitory effect was diminished when an increased proportion of Tregs was introduced. The suppressing effects of Tregs on effector T cells were attenuated by the addition of monoclonal antibody to TGF-beta, but not the one to IL-10, suggesting that TGF-beta is a major messenger involved in the immune suppressing effect of Tregs.