Peripheral blood flow cytometric analysis.
Peripheral blood was obtained 24 days before and 12 days after the initiation of anti–PD-1 treatment. Flow cytometric analysis with antibodies recognizing CD3/CD4/CD8/CD7/CD26 (catalogs 663490, 663493, 341051, 663502, and 340426, respectively; BD Biosciences) was performed on a BD FACSCanto Clinical Flow Cytometry System. Flow cytometric analysis with antibodies recognizing PD-1 (BioLegend, 329907) or mouse IgG1, κ Isotype control (BioLegend, 400122) on PBMCs before anti–PD-1 treatment was performed on a BD FACSCalibur Flow Cytometry System.