Flow Cytometry analysis.
Cell suspensions were incubated for 15 min at 4°C with Fc Block (BD Biosciences) and then stained for 30 min at 4°C with the following fluorochrome-conjugated antibodies: anti-XCR1-APC-Vio770 (clone: REA707, Miltenyi Biotec), anti-MHC-II-VioBlue (clone: REA813, Miltenyi Biotec), anti-CD11b-PerCP-Vio700 (clone: REA592, Miltenyi Biotec), anti-EpCAM-PE-Vio770 (clone: caa7-9G8, Miltenyi Biotec), anti-CD86-FITC (clone: PO3.3, Miltenyi Biotec) and anti-PD-L2-PE (clone: MIH37, Miltenyi Biotec). Cells were acquired on a MACSquant 16 flow cytometer (Miltenyi Biotec) and data were analyzed using FlowJo software using the gating strategy previously described.7 F647 fluorescence was measured using the R1 channel.