Figure 9. Cisplatin/LPS-primed DCs induced the generation of Th2-type cells and IL-10-producing Tr1 cells in vivo.

A., B., C. Spleen cells from each mouse group (i.e., mice injected with LPS-primed DCs and mice injected with Cisplatin/LPS-primed DCs) were stimulated with OVA_257-264 and OVA_323-339 for 12 h. The cells that had become stimulated with the OVA peptides were identified by intracellular cytokine staining of Ly5.1-expressing T cells. Further details regarding this in vivo experiment are provided in the Materials and Methods section. A. The frequencies of OVA-specific donor Th1 (Ly5.1⁺CD4⁺IFN-γ⁺), Th2 (Ly5.1⁺CD4⁺IL-4⁺), Th17 (Ly5.1⁺CD4⁺IL-17A⁺) and IFN-γ-producing CD8⁺ T cells (Ly5.1⁺CD8⁺IFN-γ⁺) were analyzed by flow cytometry. B. The frequencies of OVA-specific donor IL-10-producing Tr1 cells (Ly5.1⁺CD4⁺Foxp3⁻CD25⁻IL-10⁺) and Treg cells (Ly5.1⁺CD4⁺Foxp3⁺CD25⁺) were analyzed by flow cytometry. C. Culture supernatants obtained under the conditions described in the Materials and Methods section (in vivo experiment section) were harvested after 12 h, and IFN-γ, IL-4, IL-17A and IL-10 levels were analyzed by ELISA. The data are shown as the means ± SD of 4 mice per group. *p < 0.05 and***p < 0.001 compared with the group injected with LPS-primed DCs. LPS: group injected with LPS-primed DCs (○); LPS+CP: group injected with LPS/cisplatin-primed DCs (•).