Figure 4.

Reg3g inhibited dendritic cell (DC) maturation and cross-priming with CD8⁺ T cells in TME. (a) DCs from TBM were coincubated with 100 ng/ml Reg3g for 12 or 24 h. The secretions of interferon-γ (IFN-γ), tumor growth factor-β (TGF-β), interleukin-12 (IL-12), and IL-10 in DC supernatants were quantified by enzyme-linked immunosorbent assay (ELISA). (b) The expression of CD86 and MHC-II on CD11C⁺ DCs from control mice, TBM, and TBM treated with Reg3g lentiviral particles (TBM+Reg3g). In addition, culture medium from Panc02 cells (TME), and 100 ng/ml Reg3g+TME, were added to DCs from control mice. The expression of CD86 and MHC-II on DCs was determined by fluorescence-activated cell sorting (FACS). (c) T cells were cocultured with DCs at the ratio of 20 : 1, 20 : 2, and 20 : 4 for 24 h. The proliferation of T cells was then determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. (d) CD8⁺ T cells were cocultured with DCs from different groups for 24 h, and levels of granzyme B and IFN-γ in the supernatant were determined by ELISA. Data were shown as means±S.D. from at least three independent experiments. *P<0.05, **P<0.01 compared with control; ^#P<0.05, ^##P<0.01 compared with the model group