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Published ahead of print on November 20, 2003, doi:10.1165/rcmb.2003-0234OC

Am. J. Respir. Cell Mol. Biol., Volume 30, Number 5, May 2004, 736-743

A more recent version of this article appeared on May 1, 2004
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Submitted on June 23, 2003
Revised on November 20, 2003

Expression of IL-5- and GM-CSF-responsive Genes in Blood and Airway Eosinophils

Mary E Bates1, Lin Ying Liu2, Stephane Esnault3, Barbara A Stout1, Ekokobe Fonkem1, Vanderlene Kung1, Julie B Sedgwick2, Elizabeth A.B. Kelly2, Douglas M Bates4, James S Malter3, William W Busse2, and Paul J Bertics1*

1 Biomolecular Chemistry, University of Wisconsin, Madison, WI, USA, 2 Medicine, University of Wisconsin, Madison, WI, USA, 3 Pathology and Laboratory Medicine, University of Wisconsin, Madison, WI, USA, 4 Statistics, University of Wisconsin, Madison, WI, USA

* To whom correspondence should be addressed. E-mail: PBERTICS{at}FACSTAFF.WISC.EDU.

Because 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 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 {beta}, 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, 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 {beta}, 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.




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