Figure 3
CD163, CD169, SLA-DR and CD80/86 expression on PRRSV-infected moDCs. Four-color FCM including a Live/Dead discrimination dye was used to study either CD163 and CD169 or SLA-DR and CD80/86 expression in moDCs before infection (0 hpi), mock-infected (mock), PRRSV-2 infected (PRRSV-2 i.) and PRRSV-1 infected (PRRSV-1 i.) CD172ahigh-defined moDCs at 24 hpi. (A) CD172ahigh moDCs were gated (not shown) and analyzed for PRRSV-nucleoprotein expression (PRRSV-N, x-axis) versus Live/Dead staining (Near-IR, y-axis) at 0 hpi and 24 hpi for mock, PRRSV-2 and PRRSV-1 infected microcultures. Gates indicate PRRSV-N− mock-infected live moDCs (black gates), PRRSV-N− and PRRSV-N+ live moDCs from PRRSV-2 infected cultures (green gates) and PRRSV-N− and PRRSV-N+ live moDCs from PRRSV-1 infected cultures (red gates). (B) Contour plots show expression of CD163 versus CD169 (top row) and SLA-DR versus CD80/86 (bottom row) on moDCs within the respective gates shown in (A). Numbers indicate median fluorescence intensity values for CD169 and CD80/86 in the upper left corner of each contour-plot whereas numbers in the lower right corner indicate median fluorescence intensity values for CD163 and SLA-DR. Exemplary data of moDCs derived from a single animal is shown. (C) MFI values obtained for SLA-DR and CD80/86 in mock-infected moDC cultures were set to 1 (dashed lines) and ratios based on MFI values for SLA-DR (left graph) and CD80/86 expression (right graph) of N-prot.− and N-prot.+ moDCs for each genotype were calculated and displayed in the respective scatter diagrams. Each symbol represents data of moDCs from one individual animal (n = 4). Coloured bars indicate mean values. Significant differences between N-protein+ moDCs compared to N-protein− moDCs from PRRSV-2 and PRRSV-1 infected microcultures are indicated (* = p < 0.05).