Pe b7 h4

Single-cell suspensions were blocked with mouse FcR blocking reagent (Miltenyi Biotec) for 10 min at 4 °C prior to surface staining. Cell viability was assessed by Fixable viability dye eFluor 520 (eBioscience) to exclude dead cells. The following anti-mouse antibodies were used: FITC-CD11b, PE-F4/80, APC-F4/80, FITC-CD45, PerCP-eFluor 710-MHC Class II (I-A/I-E), PerCP-eFluor 710-CD3, FITC-CD3e, FITC-CD4, APC-CD4, APC-CD8a, PE-PD-L2, PE-B7-H2, PE-B7-H3, PE-B7-H4, PE-TIM-4, PE-VISTA from eBioscience; APC-CD206, PE-PD-L1, APC-CD86, PE/Cy7 Ki-67 from Biolegend; V450-CD4, BV510-CD8, PE-IFN-γ from BD. The following anti-human antibodies were used: PerCP-eFluor 710-CD3, APC-CD4, APC-CD8a from eBioscience; FITC-CD14, PE-PD-L1, APC-CD163, APC-CD86, FITC-HLA-DR, PE-Cy7-CD163, PE-CD25, PE-IFN-γ from Biolegend; FITC-CD8, PE-IL-10 from BD. For intracellular staining, cells were fixed and permeabilized with the Fixation/Permeabilization solution kit (BD). All flow cytometry data was acquired on FACSCalibur or LSRFortessa (BD, San Jose, USA) and analyzed by FlowJo V10 (TreeStar, Ashland, USA).