Time course analysis of pDC IFNα production triggered by DENV infected cells.

<p>(<b>A</b>) Quantification of IFNα in the supernatants of PBMCs and pDCs isolated from two different donors (#1 and #2, as indicated) and co-cultured with DENV infected Huh7.5.1 cells (DENV cells) or treated with 100 µl of supernatant from the latter cells (DENV SN, viral titers ≈3×10⁶ and 1×10⁶ ffu/ml for experiments with Donor #1 and #2, respectively) for the indicated time of incubation. Uninfected Huh-7.5.1 cells are referred to as (cont) cells. Error bars represent the means ± SD, n = 3. Arrows indicate results below the limit of detection of the IFNα ELISA assay (<i>i.e.</i> 12.5 pg/ml). (<b>B</b>) Quantification of IFNα-positive cells at the indicated incubation times of PBMCs or pDCs isolated from two different donors (#1 and #2, as indicated) and co-cultured with DENV infected Huh7.5.1 cells (DENV cells) or treated with 100 µl of supernatant from the latter cells (DENV SN, viral titers of ≈3×10⁶ ffu/ml). The pDC surface marker CD123 and intracellular IFNα staining were assessed by FACS analysis. Results are expressed as the percentages of IFNα positive cells in CD123 positive- and negative-populations.