Cd38 pevio770

Macrophages from peritoneal exudate derived from mice in the TGC model (0.5 × 10⁶ cells) were resuspended in FACS buffer (PBS supplemented with 0.5% bovine serum albumin and 0.1% NaN₃) and were labeled with the antibody mixes from panel 1 (CD45-APC; F4/80-Viogreen; MHCII-Vioblue; LFA-1-PeVio770; CD64-PE; CD16/32-Viobright FITC) (Miltenyi Biotec, Bergisch Gladbach, Germany) and panel 2 (CD45-APC; F4/80-Viogreen; CD64-PE; MHCII-Vioblue; CD38-PeVio770; CD206-FITC) (Miltenyi Biotec, Bergisch Gladbach, Germany) for 20 min at 4°C. Cells were then washed with FACS buffer and the relative fluorescence intensities were determined using a MACSQuant Vyb cytometer equipped with FlowLogic software (Miltenyi Biotec, Bergisch Gladbach, Germany).

1.5 × 10^{^6} of thawed PBMCs were plated in complete RPMI containing 10% human serum supplemented with 1% Penicillin–Streptomycin–Glutamin. Overnight-rested PBMCs were stained with the appropriate antibodies for 20 min at 4 °C in the dark and acquired using FACSVerse™cytometer (BD Biosciences). Dead cells were labeled using ViobilityTM Fixable Dye (Miltenyi Biotec). Antibodies used were: CD4-APC-Vio770, CD8-APC, HLA-DR-VioBlue, CD38-PE-Vio770, Granzyme B-PE and Perforin-FITC (Miltenyi Biotec). (Representative plots are shown in Supplementary Fig. 6). Data were analyzed using FlowJo 10.7.2 (BD Biosciences).