Figure 4

Induction of type I interferon by RIG-I activation prior to conditioning therapy reduces allogenicity of dendritic cells to activate allogeneic T cells. (A) Scheme of experimental setup: Balb/c mice were treated with IFNaR1 blocking antibody or were left untreated. One day later mice were treated with 3pRNA or were left untreated. One day later all mice received TBI with 9 Gy. Three days later spleens were harvested and CD11c⁺ cells were isolated and cocultured with allogeneic CD4⁺ or CD8⁺ T cells derived from C57BL/6 mice. Proliferation and IFN-γ production were analyzed on day 3 (CD8⁺ T cells) and 5 (CD4⁺ T cells) after onset of the mixed lymphocyte reaction (MLR). (B) FACS gating of T cells after MLR. CD4⁺ T cells were analyzed on day 5 after MLR. Representative data from one of five experiments. C) Left panel: Percentage of proliferated and IFN-γ⁺ cells of all CD4⁺ T cells on day 5 after onset of the MLR. Right panel: Percentage of proliferated and IFN-γ⁺ cells of all CD8⁺ T cells on day 3 after onset of the MLR. Pooled data of five independent experiments. Animal numbers per group (n) are depicted. Data were analyzed using ordinary one-way ANOVA for multiple comparisons. Significance was set at p values < 0.05, p < 0.01 and p < 0.001 and was then indicated with asterisks (*^,** and ***). Data are presented as mean ± S.E.M.