Anti-inflammatory gene therapy directed at the airway epithelium

Gene Ther. 2000 Feb;7(4):306-13. doi: 10.1038/sj.gt.3301078.

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 (NFkappaB). We have assessed the effect of adenovirus and liposome-mediated overexpression of the NFkappaB inhibitor IkappaBalpha, as well as liposome-mediated transfection with oligonucleotides resembling NFkappaB consensus binding sites (decoys) in a cystic fibrosis airway epithelial cell line (CFTE). Electrophoretic mobility shift assays (EMSA) were used to assess NFkappaB activity and secretion of the pro-inflammatory cytokine interleukin-8 (IL-8) was measured by ELISA. At a MOI of 30, Ad-IkappaBalpha significantly decreased IL-8 secretion to 60% and 43% of control unstimulated and TNF-alpha stimulated cells, respectively. At this MOI, approximately 70% of cells are transduced. EMSA showed an approximately 50% decrease in NFkappaB activation. Liposome-mediated transfection of IkappaBalpha 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-alpha stimulated IL-8 secretion was reduced by approximately 40% using this approach. EMSA confirmed a significant decrease of NFkappaB activation. Decoy oligonucleotides may be a promising approach for reduction of NFkappaB-mediated pulmonary inflammation. Gene Therapy (2000) 7, 306-313.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenoviridae / genetics
  • Cells, Cultured
  • Cystic Fibrosis / metabolism
  • Cystic Fibrosis / therapy*
  • Electrophoresis
  • Genetic Therapy / methods*
  • Genetic Vectors / genetics
  • Humans
  • Interleukin-8 / metabolism
  • Liposomes / genetics
  • Liposomes / pharmacology
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / genetics*
  • Oligonucleotides / pharmacology
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Transfection / genetics

Substances

  • Interleukin-8
  • Liposomes
  • NF-kappa B
  • Oligonucleotides