Surface staining was performed with directly conjugated monoclonal antibodies for 20 min at room temperature for human samples. The cells were washed and resuspended in PBS before flow cytometric analysis. The monoclonal antibodies used were anti‐human CD4–Percp‐Cy5.5/APC‐H7, CD8–APC (allophycocyanin)‐R700/V500, PD‐1–PE (phycoerythrin)–Cy7, TIM‐3–APC, TIGIT–BV605, 2B4–AF700, and CD160–PE (BD Bioscience San Diego, CA, USA). Intracellular staining was carried out by using a fixation/permeabilization kit (BD Bioscience) after resuspension according to the manufacturer's instructions. Ki67–PE (BD Pharmingen) was added and incubated for 20 min at room temperature.
The surfaces of mouse samples were stained with direct‐conjugated monoclonal antibodies for 30 min at 4 °C. After incubation, the cells were washed and resuspended in phosphate‐buffered saline before flow cytometry analysis. The monoclonal antibodies used were anti‐mouse CD3–Percp, CD4–APC‐H7, CD8–FITC (fluorescein), CD44–PE–Cy7, and CD62L–APC (BD Bioscience San Diego, CA, USA). Detailed antibody information was listed in TableS3 (Supporting Information).
The surfaces of mouse samples were stained with direct‐conjugated monoclonal antibodies for 30 min at 4 °C. After incubation, the cells were washed and resuspended in phosphate‐buffered saline before flow cytometry analysis. The monoclonal antibodies used were anti‐mouse CD3–Percp, CD4–APC‐H7, CD8–FITC (fluorescein), CD44–PE–Cy7, and CD62L–APC (BD Bioscience San Diego, CA, USA). Detailed antibody information was listed in Table