Pe cy7 conjugated anti tnf α

The following mAbs and reagents were used in this study: Allophycocyanin (APC)-conjugated anti-CD3, APC-conjugated anti-IL-4, APC-conjugated anti-TCRVα24-Jα18, APC-Cy7-conjugated anti-CD3, FITC-conjugated anti-CD3, FITC-conjugated anti-CD45, FITC-conjugated anti-CD56, FITC-conjugated anti-TCRγδ, FITC-conjugated anti-IFN-γ, PE-conjugated anti-TCRVα24-Jα18, PE-conjugated anti-CD45, PE-conjugated anti-CD56, PE-conjugated anti-CD69, PE-conjugated anti-IL-17A, PE-conjugated anti-lymphocyte-activation gene 3 (anti-LAG3), PE-Cy5-conjugated anti-CD161, PE-Cy7-conjugated anti-TNF-α, PerCP-conjugated anti-CD3, PerCP-conjugated anti-CD45, FITC-conjugated mouse IgG isotype, PE-conjugated mouse IgG isotype and PE-Cy7-conjugated mouse IgG isotype control (all from Becton Dickinson, San Diego, CA, USA); PE-conjugated anti-programmed death-1 (anti-PD-1; eBioscience, San Diego, CA, USA) and APC-conjugated anti-TCR Vα7.2 (BioLegend, San Diego, CA, USA). Cells were stained with combinations of appropriate mAbs for 20 min at 4°C. Stained cells were analyzed on a Navios flow cytometer using Kaluza software (version 1.5a; Beckman Coulter, Brea, CA, USA).