Anti il 18rα fitc

PBMCs were stained in 96-well U-bottom plates as described previously (6 (link)). In brief, cells were stained with fluorophore-labeled Abs to cell-surface markers, fixed, permeabilized (Cytofix/Cytoperm; BD Biosciences), and stained for intracellular molecules. The following mAbs were used: anti–CD3-V500, anti–CD56-PECy7, anti–IFN-γ–allophycocyanin, anti–CD107a-A488, anti–CD16-allophycocyanin-H7, anti–CD25-allophycocyanin-H7 (all BD Biosciences), anti–CD57-e450, anti–CD25-PErCPCy5.5, anti–CD16-allophycocyanin, anti–CD25-PE, anti–IL-18Rα–PE, anti–IL-18Rα–FITC, anti–IFN-γ–allophycocyanin-e780, anti–CD16–allophycocyanin-e780 (all e-Biosciences), anti–NKG2C-allophycocyanin, anti–NKG2C-PE (both R&D Systems), and anti–NKG2A-FITC (Miltenyi). IL-12Rβ2 Ab was conjugated using EasyLink PE-Cy5 (Abcam). Cells were acquired on an LSRII flow cytometer (BD Biosciences) using FACSDiva software. Data analysis was performed using FlowJo V10 (Tree Star). FACS gates set on unstimulated cells (medium alone or isotype controls) were applied in standard format across all samples and all conditions.