Cells were surface stained with anti-human monoclonal antibodies (mAbs) to CD3 (clone OKT3), CD14 (clone M5E2), CD19 (clone HIB19), CD161 (clone HP-3G10), TCR Vα7.2 (clone 3C10) (Biolegend, San Diego, CA, USA), CD4 (clone L200), CD8 (clone SK1), activated caspase-3 (clone C92-605), CCR6 (clone 11A9), HLA-DR (clone G46-6), Ki67 (clone B56) (BD Pharmingen, San Diego, CA, USA), CCR9 (clone 112509) (R&D, Minneapolis, MN, USA), CD38 (clone LS198.4.3) (Beckman-Coulter, Miami, FL, USA), and CD57 [clone TB01 (TB01); eBioscience, San Diego, CA, USA]. Antibodies conjugated to the following fluorochromes were used in these studies: fluorescein isothiocyanate (FITC), phycoerythrin (PE), peridinin chlorophyll protein (PerCP)-Cy5.5, PE-Cy7, energy coupled dye or PE-Texas-red conjugate (ECD), violet (V) 450 (e.g., similar to Pacific blue), brilliant violet (BV) 570, BV605, BV650, quantum dot (QD) 800, Alexa 647, allophycocyanin (APC)-Alexa 700 and APC-H7.
Culture medium consisted of RPMI 1640 (Gibco, Grand Island, NY, USA) supplemented with 100 U/ml penicillin, 100 µg/ml streptomycin, 50 µg/ml gentamicin, 2 mMl -glutamine, 2.5 mM sodium pyruvate, 10 mM HEPES buffer, and 10% heat-inactivated fetal bovine serum (R10).
Culture medium consisted of RPMI 1640 (Gibco, Grand Island, NY, USA) supplemented with 100 U/ml penicillin, 100 µg/ml streptomycin, 50 µg/ml gentamicin, 2 mM