Regulatory T cells can restrict the uncontrolled immune response and inflammation, avoiding pathologic immune injury to the host and thus playing important roles in the maintenance of immune homeostasis. Until recently, many subsets of CD4 and CD8 regulatory T cells have been reported. In this study, we identified CD11c high CD8+ T cells as a new subset of CD8+ regulatory T cells. During Listeria monocytogenes and Staphylococcus aureus infection, two subsets of CD8 T cells were classified according to the expression level of CD11c, including CD11c low CD8+ and CD11c high CD8+ T cells. CD11c low CD8+ T cells, existing during the whole period of infection, act as conventional activated T cells to kill target cells in a perforin-dependent manner. Interestingly, CD11c high CD8+ T cells appeared only at a late stage of infection, expressed relatively high CD122 and low CD69, did not secrete IFN-γ, IL-10, TGF-β, and exhibited much more potent cytotoxicity against target cells via Fas ligand–Fas pathway in an Ag-independent manner. Ligation of CD11c was important in the cytotoxicity of CD11c high CD8+ T cells. Furthermore, CD11c high CD8+ T cells could directly kill the activated CD4 T cells both in vitro and in vivo, whereas CD11c low CD8+ T cells could not. Thus, we identified an infection-induced new subset of CD11c high CD8+ regulatory T cells, which might contribute to protect host from pathological immune injure. Our results indicate that CD11c+ CD8+ T cells are constitute a heterogeneous population that can be divided further into regulatory CD11c high CD8+ T cell subset and effector CD11c low CD8+ T cell subset, thus adding insight to the role of CD8 T cells in immune response and regulation.