Fig. 3: IFNα sensitizes DCs and macrophages to TLR7/8 agonists.
a, Dendritic cells were generated from BM that was supplemented with FLT3L. Dendritic cells were treated with IFNα (500 or 5,000 U ml−1) or stimulated with IMQ (2.5 µg ml−1) for 24 h. Tlr7 mRNA expression was analyzed using RT–qPCR. (n = 6 mice per group; data are pooled from two independent experiments). b, Tlr7 mRNA expression was quantified in murine BM-derived macrophages treated as described in a. n = 6 mice per group; data are pooled from two independent experiments. c, Mice received IFNα and/or topical IMQ or oral IMQ 1 day before the analysis of TLR7 expression on myeloid cells by intracellular flow cytometry. Sample size: n = 3 mice per group from one experiment. d, Heatmap depicts TLR7 expression on myeloid cells within B16-F10 tumors. Treatments are indicated in color-coded rectangles above the heatmap, as described in c. The fold change of the geometric mean fluorescence intensity (gMFI) of TLR7 on myeloid cells of treated mice compared to the control is shown. e, Tlr7 mRNA expression was plotted against a human dendritic cell signature gene (Cd1c) in a subset of patients with melanoma included in the TCGA database. The subset consisted of patients who received IFNα before biopsy (n = 29) and a control group of patients who did not (n = 14). f, Correlation plot of Tlr7 to Cd68, a human macrophage cell signature gene, in a subset of melanoma patients from the TCGA database described in e. Bar graphs are plotted as mean ± s.e.m. Dots in a and b represent biological replicates. Correlation is shown in a xy-plot with a linear regression. Heatmaps are color-coded (blue, low; red, high value). P values were calculated by unpaired, two-tailed t-tests (d), one-way ANOVA with Tukey’s post-test (a,b) and two-tailed Pearson’s correlation test (e,f).
