Conversion of Tolerogenic CD4¯8¯ Dendritic Cells to Immunogenic Ones Inducing Efficient Antitumor Immunity

Abstract

Culturing conditions may affect dendritic cell (DC) maturation status and functional effects. We have previously demonstrated that different DC subsets play distinct roles in immune responses. The splenic CD4¯8¯ DC subset that secretes transforming growth factor (TGF)-β stimulates CD4⁺ regulatory T type 1 (Tr1) cell responses, and this leads to antitumor immune tolerance. In this study, we investigated the potential effect of culturing conditions, namely: (1) duration of culturing and (2) the dose of antigen ovalbumin(OVA) for DC pulsing, respectively, in the conversion of tolerogenic CD4¯8¯DC into immunogenic DCs. Our data showed that isolated CD4¯8¯DCs cultured for an additional 18 hours in medium containing 15–20 ng/mL granulocyte macrophage colony-stimulating factor (GM-CSF) became more mature compared to the freshly isolated CD4¯8¯DCs. When pulsed with OVA at the relatively high concentration of 1 mg/mL, but not at 0.1 mg/mL, the CD4¯8¯DCs could be converted into immunogenic CD4¯8¯DCs, which stimulated CD4⁺ T-cell differentiation into type 1 helper T (Th1) cells. Vaccination of mice with converted CD4¯8¯DCs induced strong OVA-specific cytotoxic T-lymphocyte (CTL) responses and protective immunity against OVA-expressing BL6-10OVA B16 melanoma. Taken together, our findings indicate that the conversion of DCs from a tolerogenic to an immunogenic state can be achieved by the elongation of DC culturing time in combination with a high-dose antigen for DC pulsing. Therefore, our results may have a significant impact in designing DC-based antitumor vaccines.