Anti cd8 aa700

Blood and BAL fluid immuno-staining were performed as follows: 100 μL of whole blood or BAL fluid were incubated for 10 min at room temperature in the dark with the following conjugated-monoclonal antibodies: anti-CD3-AA750, anti-CD8-AA700, anti-CD279 (PD-1)-PC7 or isotype control, anti-HLA-DR-PB or isotype control, anti-CD14-ECD and CD45-Krome Orange (Beckman Coulter). For blood samples, red-blood cells were then lysed using VersaLyse Solution (Beckman Coulter). Washed blood and BAL fluid-stained samples were immediately acquired on a 10-multicolor Navios flow cytometer and analyzed with the Kaluza 2.1 software (both from Beckman Coulter). The gating strategy is depicted in Additional file 1: Figure S1 in BAL fluid (Panel A) and blood (Panel B). HLA-DR and PD-1 quantification were expressed in percentage of positive cells or mean fluorescence of intensity (MFI).