Cytochemistry and phenotype of cells obtained from culture of freshly isolated CD15⁺CD14^- neutrophils. (A-B) Freshly isolated CD15⁺CD14^- neutrophils were cultured with GM-CSF, TNF-α, and IFN-γ for 11 days, followed by additional 7-day culture with M-CSF alone. (C-D) Freshly isolated CD15⁺CD14^- neutrophils were cultured with GM-CSF, TNF-α, IFN-γ, and IL-4 for 11 days, followed by additional 7-day culture with M-CSF alone. (A,C) Photographs of May-Grünwald-Giemsa-, MPO-, and double specific/nonspecific esterase–stained cytospin preparations; original magnification, × 400. (B,D) The expression of CD15/CD14, MPO, M-CSFR, lactoferrin, mannose receptor, and HLA-DR was analyzed using a FACSCalibur flow cytometer. In the histograms, the thick and thin lines show the expression of the indicated molecules and isotype controls, respectively. Representative data from 5 independent experiments are shown.