Flow cytometry
LYSIS
Circulating leukocyte subsets were analyzed using flow cytometry. Red blood cell lysis was performed on sodium heparinized blood using RBC lysis buffer (Thermo Fisher Scientific). After washing with PBS (pH 7.2), leukocytes were incubated with fluorochrome-labeled antibodies for 30 min on ice. After a final washing step, leukocytes were measured on a MACSQuant 16 analyzer (Miltenyi Biotec). See supplemental table 