Bronchoalveolar Lavage Cytokine Profiling

Mice were sacrificed by intraperitoneal (i.p.) administration of 90 mg/kg of body weight sodium pentobarbital. Bronchoalveolar lavage fluid (BALF) was obtained by lavaging the lungs with 3 mM EDTA in PBS (11 (link)), and cellular composition was determined by hemocytometer cell counts and differential counts of cytospins after staining with Quick-Diff solution (Siemens; Medical Solutions Diagnostics, Tarrytown, NY). Cell-free BALF was used to determine levels of IL-13 (4 to 500 pg/ml), IFN-β (1.9 to 500 pg/ml), and IFN-γ (15.6 to 1,000 pg/ml) using enzyme-linked immunosorbent assay (ELISA) kits (Ready-Set-Go; eBioscience, San Diego, CA, or BioLegend). Results are from 5 mice per group (biological replicates) and 3 technical replicates per mouse.