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Published ahead of print on November 20, 2003, doi:10.1165/rcmb.2003-0234OC
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American Journal of Respiratory Cell and Molecular Biology. Vol. 30, pp. 736-743, 2004
© 2004 American Thoracic Society
DOI: 10.1165/rcmb.2003-0234OC

Expression of Interleukin-5– and Granulocyte Macrophage–Colony-Stimulating Factor–Responsive Genes in Blood and Airway Eosinophils

Mary E. Bates, Lin Ying Liu, Stephane Esnault, Barbara A. Stout, Ekokobe Fonkem, Vanderlene Kung, Julie B. Sedgwick, Elizabeth A. B. Kelly, Douglas M. Bates, James S. Malter, William W. Busse and Paul J. Bertics

Departments of Biomolecular Chemistry, Medicine, Pathology, and Laboratory Medicine and Statistics, University of Wisconsin–Madison, Madison, Wisconsin

Address correspondence to: Paul J. Bertics, Ph.D., Dept. of Biomolecular Chemistry, University of Wisconsin, 1300 University Ave., Madison, WI 53706. E-mail: pbertics{at}facstaff.wisc.edu

Because interleukin (IL)-5 family cytokines are critical regulators of eosinophil development, recruitment, and activation, this study was initiated to identify proteins induced by these cytokines in eosinophils. Using oligonucleotide microarrays, numerous transcripts were identified as responsive to both IL-5 and granulocyte macrophage–colony-stimulating factor (GM-CSF), but no transcripts were markedly affected by one cytokine and not the other. Expression of several gene products were seen to be increased following in vitro stimulation of human blood eosinophils, including the IL-3 receptor {alpha} subunit, lymphotoxin ß, Pim-1, and cyclin D3. Given that eosinophils recovered from the bronchoalveolar lavage fluid of allergic patients after antigen challenge are exposed to IL-5 or GM-CSF in the airway prior to isolation, the hypothesis was tested that selected IL-5– and GM-CSF–responsive genes are upregulated in airway eosinophils relative to the expression in blood cells. Airway eosinophils displayed greater cell surface expression of the IL-3 receptor {alpha} subunit, CD44, CD25, and CD66e, suggesting that these proteins may be markers of eosinophil activation by IL-5 family cytokines in airway eosinophils. Other genes that were induced by both IL-5 and GM-CSF showed protein expression at similar or decreased levels in airway eosinophils relative to their circulating counterparts (i.e., lymphotoxin ß and CD24). These studies have identified several transcriptional targets of IL-5 and GM-CSF in human eosinophils and suggest that a number of protein products are critical to the responsiveness of airway eosinophils.

Abbreviations: bronchoalveolar lavage, BAL • biliary glycoprotein a, BGPa • interleukin, IL • granulocyte macrophage–colony-stimulating factor, GM-CSF • lymphotoxin, LT • Model-Based Expression Index, MBEI • median channel fluorescence, MCF • segmental bronchoprovocation with antigen, SBP-Ag • tumor necrosis factor, TNF




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