Lungs were perfused with PBS containing EDTA (0.5 mM), minced, and digested in collagenase IV (5 mg/ml) and DNase I (23 (link)). Cells were filtered and washed with RBC lysis buffer (BD Biosciences, Franklin Lakes NJ) and kept on ice in media containing 10% serum. Dead cells were stained with Pac-Orange Live/Dead fixable dead staining dye (Invitrogen). Lung cells were then stained with fluorescent-labeled antibodies against various leukocyte surface markers (CD45, CD11b, CD11c, F4/80, CD103, MHCII, CD86, Clec9a, CD3ε, TCRβ, CD4, CD69, IFN-γ). Appropriate isotype-matched controls were used in all experiments. Antibodies were purchased from EBiosciences (San Diego, CA) or Biolegend (San Diego, CA). Cells were fixed and analyzed on a Canto2 (Becton-Dickinson, San Jose, CA) or FACSAria II (BD Biosciences) flow cytometer. Results were analyzed using FlowJo software (Tree Star, Ashland, OR). For analysis of intracellular IL-12 or IFN-γ, fresh aliquots of digested lung tissue were stimulated for 5 hours at 37°C, with Cell-stimulation Cocktail Buffer (40.5 μmol/L phorbol 12-myristate 13-acetate [PMA], 670 μmol/L ionomycin, 5.3 mmol/L brefeldin A, and 1 mmol/L monensin; eBioscience), fixed, permeabilized with Cell Permeabilization Buffer (eBioscience) and incubated with anti-mouse IL-12 clone C17.8 (eBioscience) or anti-mouse IFN-γ clone XMG1.2 (Biolegend).