Cd94 apc

Flow cytometry was performed on 1E6 stained cells from each enriched fraction, including a non-enriched sample from each tonsil for comparison. Surface and intracellular flow cytometry was performed as previously described12 (link) on a Becton Dickenson LSR II flow cytometer using FACS Diva analysis software (BD Biosciences). The following commercially available antibodies were used: CD8-PE, CD19-PE, CD19-FITC, CD94-APC, CD3-APC-H7, and IFN-γ-BV421 (BD Biosciences); CD3-PerCP-Vio700, CD14-PerCP-Vio700, CD117-PE-Vio770, CD3-VioGreen, and CD14-VioGreen (Miltenyi Biotec); and IL-22-PE (R&D Systems). Cytofix/Cytoperm and PermWash reagents (BD Biosciences) were used to perform intracellular flow cytometry as previously described12 (link).

The Monoclonal antibodies (mABs) used for phenotypic analysis included Fluorescein isothiocyanate (FITC)-, Phycoerythrin (PE)-, Phycoerythrin Cyanine 5.5 and allophycocyanin (APC)-labeled anti-CD45, anti-CD3, anti-CD56, NKp30, NKp44 (Beckman Coulter, Brea, CA, USA), anti-CD16, NKp46, NKG2A (R&D System), anti-CD94, anti-CD69, anti-CD18, anti-CD49, anti-CD62L, anti-CD38 and CXCR4 (BD Pharmingen, San Jose, CA, USA). Phenotype evaluation was performed by direct immunofluorescence according to previously reported methods [29 (link)]. NK cells recovered from immunodeficient NOD/SCID gamma (NSG) mice were stained with anti-mouse CD45-APC-Cy7, anti-human CD45-PE-Cy7, CD3-PerCP-Cy5.5, CD56-FITC (Biolegend, San Diego, CA, USA) and CD94-APC (BD).