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. 2015 Feb 5;45(4):1159–1169. doi: 10.1002/eji.201445132

Figure 4.

Figure 4

The synergistic effect of the combination of K3 CpG and cGAMP on antigen-specific IFN-γ induction is dependent on IRF3/7, STING, MyD88, IL-12, and type-I IFN signaling. (A) WT, Tmem173gt, IRF3/7 DKO, MyD88 KO, and IFNAR KO C57BL/6J mice (n ≥ 3) were immunized with OVA and K3 CpG, cGAMP, or K3 + cGAMP at days 0 and 10, via the i.m. route. On day 17, OVA-specific serum IgG2c and IgG1 were measured by ELISA. Each symbols represent an individual mouse and data are representative of two independent experiments and are shown as the mean + SD of biological replicates from one experiment, representative of two performed. *p < 0.05; **p < 0.01; ***p < 0.001 (one-way ANOVA with Bonferroni's multiple comparison test). (B) Spleen cells from immunized mice were stimulated with OVA for 48 h. Production of IFN-γ and IL-13 were measured by ELISA. Data are representative of two independent experiments and are shown as the mean + SD of biological replicates from one experiment, representative of two performed. *p < 0.05; **p < 0.01; ***p < 0.001 (one-way ANOVA with Bonferroni's multiple comparison test). (C) IL-12p40 +/− and −/− C57BL/6J mice were immunized with OVA and K3 CpG, cGAMP, or K3 CpG + cGAMP at days 0 and 10, via the i.m. route. On day 17, OVA-specific serum IgG2c and IgG1 were measured by ELISA. Data are representative of two independent experiments and are shown as the mean + SD of biological replicates from one experiment, representative of two performed. *p < 0.05 (Mann–Whitney U-test). (D) Spleen cells were stimulated with OVA protein for 48 h. Production of IFN-γ was measured by ELISA. Data are representative of two independent experiments and are shown as the mean + SD of biological replicates from one experiment, representative of two performed. *p < 0.05 (Mann–Whitney U-test).