title: Functionally competent eosinophils differentiated ex vivo in high purity from normal mouse bone marrow.
abstract: We have devised an ex vivo culture system which generates large numbers of eosinophils at high purity (>90%) from unselected mouse bone marrow progenitors. In response to 4 days of culture with recombinant mouse FLT3-L and recombinant mouse stem cell factor followed by recombinant mouse IL-5 alone thereafter, the resulting bone marrow-derived eosinophils (bmEos) express immunoreactive major basic protein, Siglec F, IL-5R alpha-chain, and transcripts encoding mouse eosinophil peroxidase, CCR3, the IL-3/IL-5/GM-CSF receptor common beta-chain, and the transcription factor GATA-1. BmEos are functionally competent: they undergo chemotaxis toward mouse eotaxin-1 and produce characteristic cytokines, including IFN-gamma, IL-4, MIP-1alpha, and IL-6. The rodent pathogen pneumonia virus of mice replicates in bmEos and elevated levels of IL-6 are detected in supernatants of bmEos cultures in response to active infection. Finally, differentiating bmEos are readily transfected with lentiviral vectors, suggesting a means for rapid production of genetically manipulated cells.
authors.affiliation: Eosinophil Biology Section, Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA. kdyer@niaid.nih.gov
authors.forename: Kimberly D
authors.initials: KD
authors.lastname: Dyer
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Moser
authors.forename: Meggan
authors.initials: M
authors.lastname: Czapiga
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Siegel
authors.forename: Caroline M
authors.initials: CM
authors.lastname: Percopo
authors.forename: Helene F
authors.initials: HF
authors.lastname: Rosenberg