Figure 1

Gating strategy for identification of respiratory leukocytes, neutrophils, eosinophils and dendritic cells. Cells were gated based on scatter light (FSC, SSC) characteristics and respiratory leukocytes were identified by CD45 expression (A, B). Out of the CD45+ cells, neutrophils were identified by Gr1^brightCD11b^bright expression. Subsequently, out of the neutrophil negative fraction, eosinophils were identified by SiglecF⁺MHC-class-II^neg (I-A/I-E^neg) expression. Out of the neutrophil and eosinophil negative fraction macrophages were identified as SiglecF⁺⁺F4/80⁺ double positive cells (A). Total dendritic cells were identified as CD11c⁺ SiglecF^neg CD49b^neg respiratory leukocytes (B).