| Autor: |
Duncan, James E., Whitsett, Jeffrey A., Horowitz, Ann D. |
| Zdroj: |
Human Gene Therapy; March 1997, Vol. 8 Issue: 4 p431-438, 8p |
| Abstrakt: |
ABSTRACTCationic lipid-mediated transfection of the alveolar epithelium in vivowill require exposure of plasmid DNA and cationic lipids to endogenous surfactant lipids and proteins in the alveolar space. Effects of pulmonary surfactant and of surfactant constituents on transfection in vitroof two respiratory epithelial cell lines (MLE-15 and H441) with a plasmid encoding the luciferase reporter gene were studied using two cationic lipid formulations: 1,2-dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide/cholesterol (DMRIE/C) and 1,2-dioleoyl-3-trimethylammonium propane/dioleoyl phosphatidylethanolamine (DOTAP/DOPE). Gene expression, as assessed by luciferase activity, decreased as increasing concentrations of natural surfactant were added to cationic lipidDNA complexes. Incorporation of phospholipids DOPC/DOPG or surfactant proteins SP-B or SP-C in the cationic lipid formulation inhibited transfection. A fluorescent lipid mixing assay was used to determine the effects of surfactant proteins SP-B and SP-C on mixing between cationic lipidDNA complexes and surfactant lipid vesicles. Mixing between DOPC/DOPG vesicles and cationic lipidDNA complexes in the absence of added proteins amounted to 1020%. Addition of SP-B or SP-C increased the mixing of DOPC/DOPG vesicles with DOTAP/DOPEDNA complexes, but not DMRIECDNA complexes. These results demonstrate that pulmonary surfactant lipids and proteins inhibit transfection with cationic lipidDNA complexes in vitro, and may therefore represent a barrier to gene transfer in the lung. |
| Databáze: |
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