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Functionally Competent Eosinophils Differentiated Ex Vivo in High Purity from Normal Mouse Bone Marrow¹

Kimberly D. Dyer^2,*, Jennifer M. Moser^*, Meggan Czapiga, Steven J. Siegel^*, Caroline M. Percopo^* and Helene F. Rosenberg^*

^* Eosinophil Biology Section, Laboratory of Allergic Diseases and Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892

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 -chain, and transcripts encoding mouse eosinophil peroxidase, CCR3, the IL-3/IL-5/GM-CSF receptor common β-chain, and the transcription factor GATA-1. BmEos are functionally competent: they undergo chemotaxis toward mouse eotaxin-1 and produce characteristic cytokines, including IFN-, IL-4, MIP-1, 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.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work was funded by the NIAID Division of Intramural Research.

² Address correspondence and reprint requests to Dr. Kimberly D. Dyer, Eosinophil Biology Section, Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 10 Center Drive MSC 1883, Building 10, Room 11C216, Bethesda, MD 20892-1883. E-mail address: kdyer{at}niaid.nih.gov

³ Abbreviations used in this paper: βc, common β-chain; PVM pneumonia virus of mice; bmEos, bone marrow-derived eosinophil; SCF, stem cell factor; rm, recombinant mouse; m, mouse; EPO, eosinophil peroxidase; MBP, major basic protein; MPO, myeloperoxidase; DAPI, 4',6-diamidino-2-phenylindole; HI, heat inactivated; RSV, respiratory syncytial virus.