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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-
and interleukin-1
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-- and interleukin-1-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-; interleukin-1; airway disease
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