Anti tim3 percp cy5

The staining antibodies were anti-CD4-APC, anti-TIM3-PerCP-Cy5.5, anti-IFN-γ-PerCP, anti-IL-2-PE, and anti-TNF-α-FITC antibodies (Biolegend, USA), and anti-CD8-FITC (BD Biosciences, CA, USA).

T cells were stained with the following fluorochrome-conjugated mouse monoclonal anti-human antibodies for 20 minutes at room temperature: anti-CD25 FITC (BioLegend, catalog no. 302604, RRID:AB_314274), anti-CD38 FITC (BioLegend, catalog no. 303504, RRID:AB_314356), anti-TIM3 PerCP-Cy5.5 (BioLegend, catalog no. 345015), anti-CD4 Pacific Blue (BioLegend, catalog no. 317429), anti-CD45RO Brilliant Violet 605 (BioLegend, catalog no. 304238), anti-TIGIT Brilliant Violet 605 (BioLegend, catalog no. 372711), anti-PD1 Brilliant Violet 650 (BioLegend, catalog no. 329949), anti-CD3 Brilliant Violet 711 (BioLegend, catalog no. 317328), anti-CD101 PE (BioLegend, catalog no. 331011, RRID:AB_2716106), anti-LAG3 PE Dazzle (BioLegend, catalog no. 369331), anti-CD62 L PE Dazzle (BioLegend, catalog no. 304842), anti-CD8 PE-Cy7 (BioLegend, catalog no. 301012), and the anti-BCMA CAR idiotype APC (Allogene Therapeutics). T cells were then washed with PBS and stained with the e780 fixable viability dye (Thermo Fisher, catalog no. 65–0865–14) for 30 minutes at 4°C. All the samples were fixed using BD stabilizing fixative (BD Biosciences, catalog no. 339860) for further FACS analysis.