BALF was obtained by washing the lungs twice with 1 mL of sterile normal saline under anesthesia and centrifuging the obtained solution at 540×g for 5 min at 4 °C. The pellets were treated with red blood cell lysis buffer (Biolegend, San Diego, CA, USA) and resuspended in 1 mL of PBS (−), following which the number of cells were counted. Cytospin preparations were stained with Diff-Quik (International Reagents Corporation, Osaka, Japan) and examined under an optical microscope.

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