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  • Inherited Disease
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Anti-inflammatory gene therapy directed at the airway epithelium

Gene Therapy volume 7, pages 306–313 (2000)Cite this article

Abstract

Cystic fibrosis (CF) is characterised by chronic airway inflammation. Pro-inflammatory mediators in the lung are regulated by the transcription factor nuclear factor kappa B (NFκB). We have assessed the effect of adenovirus and liposome-mediated overexpression of the NFκB inhibitor IκBα, as well as liposome-mediated transfection with oligonucleotides resembling NFκB consensus binding sites (decoys) in a cystic fibrosis airway epithelial cell line (CFTE). Electrophoretic mobility shift assays (EMSA) were used to assess NFκB activity and secretion of the pro-inflammatory cytokine interleukin-8 (IL-8) was measured by ELISA. At a MOI of 30, Ad-IκBα significantly decreased IL-8 secretion to 60% and 43% of control unstimulated and TNF-α stimulated cells, respectively. At this MOI, approximately 70% of cells are transduced. EMSA showed an approximately 50% decrease in NFκB activation. Liposome-mediated transfection of IκBα did not reduce IL-8 secretion, probably due to low transfection efficiency (approximately 5% of cells). Liposome-mediated transfection of CFTE cells with rhodamine-labeled decoy oligonucleotides indicated a transfection efficiency close to 100%. TNF-α stimulated IL-8 secretion was reduced by approximately 40% using this approach. EMSA confirmed a significant decrease of NFκB activation. Decoy oligonucleotides may be a promising approach for reduction of NFκB-mediated pulmonary inflammation.

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Acknowledgements

We thank Patrick Baeuerle for the IκBα cDNA, Judy St George for the Adeno-βgal, Ty Pitt for the Pseudomonas bacteria, Dieter Gruenert for the CFTE cells and Naruya Tomita for advice in using NFκB decoys. The study was funded by the Wellcome Trust, the Cystic Fibrosis Trust, by NIH grant DK44935 (LH) and a Wellcome Senior Clinical Fellowship (EWFWA).

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Authors and Affiliations

  1. Department of Gene Therapy, National Heart and Lung Institute, London, UK

    U Griesenbach, P Scheid, E Hillery, D M Geddes & E W F W Alton

  2. Department of Vascular Biology and Thrombosis Research, University of Vienna, Vienna, Austria

    R de Martin

  3. Laboratory of Drug Targeting, University of Pittsburgh, PA, USA

    L Huang

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  1. U Griesenbach
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Griesenbach, U., Scheid, P., Hillery, E. et al. Anti-inflammatory gene therapy directed at the airway epithelium. Gene Ther 7, 306–313 (2000). https://doi.org/10.1038/sj.gt.3301078

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