Associate Scientist, Inflammation Immunobiology Section, LAD
Eosinophil researchers are handicapped by the relative rarity of eosinophils. We have developed a method for generating eosinophils from unselected bone marrow in large numbers and high purity. This protocol has facilitated eosinophil research worldwide. To generate bone marrow eosinophils (bmEos), bone marrow cells are collected from the femur and the tibia, the red blood cells removed, and the remaining cells put into culture with two cytokines that support the maintenance and proliferation of hematopoietic stem cells. After four days, the cells are cultured in interleukin-5, a cytokine known to support the differentiation of eosinophils. By day 12, the cultures approach 100 percent purity. Phenotypically, these cells look like eosinophils and produce typical eosinophil transcripts, express typical surface markers, release typical cytokines, and exhibit chemotaxis toward eotaxin. We have shown that this method works well in several strains of mice.
The Inflammation Immunobiology Section is interested in the role that eosinophils play in mediating host defense against viral pathogens. We have used bmEos to demonstrate that eosinophils are a target of pneumonia virus of mice (PVM). Infection with PVM stimulated the release of proinflammatory cytokines in response to the replication of the virus, and MyD88, an adaptor protein for toll-like receptors, mediated this effect.
In collaboration with other eosinophil researchers, we have examined the response of bmEos to platelet activating factor (PAF), a known secretagogue for human eosinophils. PAF stimulated the release of the granule protein eosinophil peroxidase from mouse bmEos. Unexpectedly, PAF-stimulated degranulation was independent of the known PAF receptor, suggesting the existence of a second PAF receptor.
We have used bmEos in vivo to answer questions relating to eosinophil migration toward eotaxin in models of eosinophil-deficient mice as well as in wild-type mice. We have also generated a flow-based method that will allow us to identify resting and activated eosinophils in mouse tissues.
Dr. Dyer received her Ph.D. from Georgetown University in Washington, DC. Prior to joining the Laboratory of Allergic Diseases in 2003 as a staff scientist, Dr. Dyer held a similar position in the Laboratory of Host Defenses at the National Institutes of Health.
Percopo CM, Dyer KD, Ochkur SI, Luo JL, Fischer ER, Lee JJ, Lee NA, Domachowske JB, Rosenberg HF. Activated mouse eosinophils protect against lethal respiratory virus infection. Blood. 2014 Jan 30;123(5):743-52.
Dyer KD, Garcia-Crespo KE, Percopo CM, Sturm EM, Rosenberg HF. Protocols for identifying, enumerating, and assessing mouse eosinophils. Methods Mol Biol. 2013;1032:59-77.
Sturm EM, Dyer KD, Percopo CM, Heinemann A, Rosenberg HF. Chemotaxis of bone marrow derived eosinophils in vivo: A novel method to explore receptor-dependent trafficking in the mouse. Eur J Immunol. 2013 Aug;43(8):2217-28.
Rosenberg HF, Dyer KD, Foster PS. Eosinophils: changing perspectives in health and disease. Nat Rev Immunol. 2013 Jan;13(1):9-22.
Dyer KD, Garcia-Crespo KE, Glineur S, Domachowske JB, Rosenberg HF. The pneumonia virus of mice (PVM) model of acute respiratory infection. Viruses. 2012 Dec;4(12):3494-510.
Dyer KD, Garcia-Crespo KE, Killoran KE, Rosenberg HF. Antigen profiles for the quantitative assessment of eosinophils in mouse tissues by flow cytometry. J Immunol Methods. 2011 Jun 30;369(1-2):91-7.
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Last Updated May 27, 2014