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Am J Physiol Lung Cell Mol Physiol 280: L841-L852, 2001;
1040-0605/01 $5.00
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Vol. 280, Issue 5, L841-L852, May 2001

EDITORIAL FOCUS
Identification of genes induced by inflammatory cytokines in airway epithelium

Paul Cooper, Suzanne Potter, Beatrice Mueck, Shida Yousefi, and Gabor Jarai

Novartis Horsham Research Centre, Horsham RH12 5AB, United Kingdom

Epithelial cells lining the airways are thought to play a prominent role in respiratory diseases. We utilized cDNA representational difference analysis to identify the genes in which expression is induced by the proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1beta in primary human bronchial epithelial cells and hence are relevant to airway inflammation. Hybridization of the subtraction product to arrayed cDNAs indicated that known tumor necrosis factor-alpha - and interleukin-1beta -inducible genes such as B94, Zfp36, and regulated on activation normal T cell expressed and secreted were represented, confirming the success of the subtraction experiment. A 1,152-clone library potentially representing genes with higher transcript levels in cytokine-treated human bronchial epithelial cells was generated and sequenced. Sequence similarity searches indicated that these clones represented 57 genes of known function, 1 gene of unknown function, 6 expressed sequence tags, and 2 novel sequences. The expression of 19 of these clones was studied by a combination of Northern blotting and RT-PCR analyses and confirmation of differential expression for 10 known genes, 2 expressed sequence tags, and a novel sequence not represented in any of the public databases was obtained. Thus cDNA representational difference analysis was utilized to isolate known and novel differentially expressed genes, which putatively play a role in airway inflammation.

representational difference analysis; human bronchial epithelial cells; tumor necrosis factor-alpha ; interleukin-1beta ; airway disease


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