Inhibitory effects of tumor necrosis factor-alpha on cationic lipid-mediated gene delivery to airway cells in vitro.
Cationic liposomes have been used successfully for DNA delivery to airway cells in vitro and are being tested in human clinical trials for their efficacy in cystic fibrosis transmembrane conductance regulator (CFTR) gene delivery in cystic fibrosis patients. While cationic liposomes are effective for transfection of airway cells in culture, they have not been effectively used for gene delivery to human airway cells in vivo. Several barriers in cystic fibrosis lungs, including increased amounts of mucus, phagocytic cell activity and cytokine-rich milieu caused by inflammation, may cause inhibition of gene transfection. As presented in this paper, we examined the effects of inflammatory cytokines on cationic lipid-mediated transfection of model airway cells. The results of these experiments indicate that tumor necrosis factor (TNF)-alpha dramatically inhibits Lipofectin-mediated transfection efficiency of H441 cells. Addition of anti-TNF-alpha neutralizing antibody results in recovery of efficiency. Results of temporal studies are consistent with the concept that TNF-alpha reduces transfection efficiency by a mechanism(s) other than or in addition to gene expression. These results are corroborated by fluorescence microscopic experiments which demonstrate that endocytosis of lipoplex is altered in the presence of TNF-alpha.
Baatz
J E
JE
Department of Pediatrics, Medical University of South Carolina, Charleston 29425, USA. baatzje@musc.edu
Zou
Y
Y
Korfhagen
T R
TR
D002478:Cells, Cultured; D018920:Coculture Techniques; D004247:DNA / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D016209:Interleukin-8 / Q000494:pharmacology; D008081:Liposomes; D008156:Luciferases / Q000235:genetics; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010714:Phosphatidylethanolamines / Q000737:chemistry* / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D015850:Interleukin-6; D016209:Interleukin-8; D008081:Liposomes; D010714:Phosphatidylethanolamines; D014409:Tumor Necrosis Factor-alpha; C020888:1,2-dielaidoylphosphatidylethanolamine; D007371:Interferon-gamma; D004247:DNA; D008156:Luciferases