Isolation and Phenotyping of Peritoneal T and NK Cells

Briefly, peritoneal fluid was collected in cold FACS and processed into single cell suspension by using RBC lysis buffer (Sigma-Aldrich, USA) for about 60 seconds and then washed with cold FACS. Then all samples were filtered with 70µm strainer (ThermoFisher, USA), centrifuged at 4°C, 1000 RPM for 10 minutes and then the cell pellets were resuspended in cold FACS. To determine the T cell and natural killer (NK) subset, isolated cells were stained with anti-CD3-APC and NK1.1-BB515 antibodies (Biolegend, USA), respectively. Finally, the stained samples were analyzed by using BD-FACS Celesta flow cytometry system (BD, USA) and acquired data were visualized by using built-in Diva software (BD, USA) as described before (41 (link)).