Anti-inflammatory gene therapy directed at the airway epithelium
| Autor: | P. Scheid, Leaf Huang, Uta Griesenbach, Ewfw Alton, Elizabeth Hillery, Duncan M. Geddes, R de Martin |
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| Rok vydání: | 2000 |
| Předmět: |
Electrophoresis
medicine.medical_specialty Cystic Fibrosis Genetic enhancement medicine.medical_treatment Genetic Vectors Oligonucleotides Inflammation Biology Transfection Adenoviridae Internal medicine Genetics medicine Humans Secretion Molecular Biology Cells Cultured Reverse Transcriptase Polymerase Chain Reaction Interleukin-8 NF-kappa B Genetic Therapy Molecular biology IκBα Cytokine Endocrinology Cell culture Liposomes Molecular Medicine Respiratory epithelium medicine.symptom |
| Zdroj: | Gene Therapy. 7:306-313 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/sj.gt.3301078 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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