Cell Research, 15(2):111-119, Feb 2005

Down-regulation of IL-8 expression in human airway epithelial cells through helper-dependent adenoviral-mediated RNA interference

Huibi CAO^1,4, Anan WANG¹, Bernard MARTIN¹, David R. KOEHLER^1,4, Pamela L. ZEITLIN⁵, A. Keith TANAWELL^1,2,3, Jim HU^1,2,4,*

¹Programme in Lung Biology Research and the Canadian Institutes of Health Research Group in Lung Development, Hospital for Sick Children, Toronto, Canada M5G 1X8.
²Department of Paediatrics, University of Toronto, Toronto, Canada M5S 1A1
³Department of Physiology, University of Toronto, Toronto, Canada M5S 1A1
⁴Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada M5S 1A1
⁵Departments of Pediatrics, The Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.

Received, Dec 1, 2004   Revised, Jan 5, 2005  Accepted, Jan 9, 2005

Correspondence: Jim HU 01-416-813-6412 (phone) 01-416-813-5771 (fax) jhu@sickkids.on.ca

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Abstract

Interleukin (IL)-8 is a potent neutrophil chemotactic factor and a crucial mediator in neutrophil-dependent inflammation. Various cell types produce IL-8, either in response to external stimuli such as cytokines or bacterial infection, or after malignant transformation. Anti-IL-8 strategies have been considered for anti-inflammatory therapy. In this paper we demonstrate that the RNA interference technique can be used to efficiently down-regulate IL-8 protein expression in airway epithelial cells. We used a helper-dependent adenoviral vector to express a small hairpin (sh)RNA targeting human IL-8 in cultured airway epithelial cells (IB3-1, Cftr^-/-; C38, Cftr-corrected) stimulated with TNF-¦Á, IL-1¦Â or heat-inactivated Burkholderia cenocepacia. Stimulated IL-8 expression in IB3-1 and C38 cells was significantly reduced by shRNA expression. The shRNA targeting IL-8 had no effect on the activation of NF-¦ÊB, or on the protein levels of IkB or IL-6, suggesting that this anti-IL-8 strategy was highly specific, and therefore may offer potential for the treatment of inflammatory diseases.

Keywords: Interleukin-8, RNA interference, helper-dependent adenoviral vector, inflammation, chemokine, neutrophil, cystic fibrosis.