Fig. 3

Activation of dendritic cells by LOAd viruses. Monocytes were isolated from peripheral blood mononuclear cells and cultured with GM-CSF/IL-4 to induce immature dendritic cells (DCs). DCs were infected with LOAd(-), LOAd703 or LOAd732, or left untreated. 48 h post infection, cell culture supernatants and cells were harvested and analyzed for DC maturation with flow cytometry and multiplex analysis. Flow cytometry results in A and B are displayed as relative mean fluorescence intensity (RMFI) compared to matched isotype control antibodies. DCs were gated on CD1a expression and the MFI of the respective marker was determined for CD1a + cells. In A, the transgene expression in DCs is shown (A) and B displays DC activation markers (B). In C, cytokine and chemokine levels in the culture supernatants are shown in pg/mL (C). D displays the ratio of IL-12 to IL-10 in culture supernatants (D). Bar graphs display the mean ± SD of seven biological repeats. Statistical differences between LOAd-infected and untreated cells were determined with Kruskal–Wallis test followed by Dunn’s multiple comparison test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001)