The cells yielded from the BAL and lung were stained using fluorescent labeled antibodies at 4°C in the dark for 20 minutes, washed with PBS, and then examined on the BD Biosciences (San Jose, CA) LSR II platform in the BRI flow cytometry core.
Leukocytes were stained for CD11b, CD11c, CD45.2, CD64, CD103, Ly6C, Ly6G, MHC-II, SiglecF (all Biolegend), CD253 (TRAIL), and Viability (eBioscience). The eBioscience (ThermoFisher Scientific, Foster City, CA)) Annexin V Apoptosis Detection Kit was used in accordance with the manufacturer’s instructions to determine which of the cells that absorbed the Viability dye had undergone a programmed cell death process. (SeeSupplemental Fig.1 for the complete staining and pictorial gating scheme.)
Leukocytes were stained for CD11b, CD11c, CD45.2, CD64, CD103, Ly6C, Ly6G, MHC-II, SiglecF (all Biolegend), CD253 (TRAIL), and Viability (eBioscience). The eBioscience (ThermoFisher Scientific, Foster City, CA)) Annexin V Apoptosis Detection Kit was used in accordance with the manufacturer’s instructions to determine which of the cells that absorbed the Viability dye had undergone a programmed cell death process. (See