Anti cd19 apc cy7

SKW-3.β₂m^null.CD8αα or SKW-3.β₂m ^null.CD8αβ (10⁵ per sample) were stained with MR1 or MHC-I tetramers in PBS + 2% FBS for 20 min at 4°C in the dark. Cells were washed with PBS + 2% FBS and resuspended in a surface antibody stain consisting of anti-CD3-BV421 (#562426, UCHT1; BD Horizon), anti-CD8α-BUV805 (#564912, SK1; BD Horizon), anti-CD8β-APC (#641058, 2ST8.5H7; BD FastImmune), and LIVE/DEAD fixable Near-IR dead cell stain (#L10119; Thermo Fisher Scientific) for a further 20 min at 4°C in the dark. Cells were washed twice with PBS + 2% FBS and data were acquired using a BD LSR Fortessa (BD Biosciences). PBMCs were stained with MR1 tetramers as described in (Souter et al., 2019 (link)). In brief, PBMCs (10⁷ per sample) were stained with MR1 tetramer in PBS + 2% FBS for 30 min at room temperature in the dark, washed with PBS + 2% FBS, and stained with surface antibodies anti-CD3-BV421, anti-CD19-APC-Cy7 (#302218, HIB19; Biolegend), anti-CD14-APC-Cy7 (#557831, MφP9; BD Pharmingen), anti-CD8α-BUV805, anti-CD8β-APC, anti-CD161-PE-Vio770 (#130-113-597, REA631; Miltenyi Biotec), anti-CD4-AF700 (#557922, RPA-T4; BD Pharmingen), and LIVE/DEAD fixable Near-IR dead cell stain for 20 min at 4°C. Cells were washed twice and resuspended in PBS + 2% paraformaldehyde (PFA) before data acquisition on a BD LSR Fortessa.