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 Gr1brightCD11bbright expression. Subsequently, out of the neutrophil negative fraction, eosinophils were identified by SiglecF+MHC-class-IIneg (I-A/I-Eneg) 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+ SiglecFneg CD49bneg respiratory leukocytes (B).