| Abstract
| - Objective: A novel hemagglutinating virus of Japan (HVJ)-liposome-mediated gene transfer system has been shown to have benefits of a high efficiency of transfection and low immunogenicity. The aims of this study were to determine the effect of re-transfection of the HVJ-liposome system via the airway, and to quantify the distribution of gene expression between transtracheal and transplantation approaches. Methods: β-Galactosidase (β-gal) plasmid DNA was introduced into lung tissues using the HVJ-liposome method. Two groups of Sprague-Dawley (SD) rats received intratracheal instillation of 10μg of the β-gal gene, once on Day 0 in 1 group (Group Tb-1, n=4) and 3 times on Days 0-2 in another (Group Tb-3, n=4). In a third group of SD rats (Group Tx, n=5), an orthotopic left lung transplantation was performed after the donor lung was flushed with an HVJ-liposome complex solution and preserved for 1h. Gene expression and distribution in lung tissue was then quantified by counting the X-gal stained cells. Results: Both the transtracheal and transplantation approaches resulted in low levels of transfection in the vascular endothelial cells (0.2±0.1 and 4.0±1.8%), respectively, but a moderate degree of transfection to the airway (11.0±7.1 and 28.0±20.7%) and alveolar cells (3.0±1.8 and 6.0±3.6%). Three repetitive injections via the airway increased gene expression in airway epithelial cells of 41.0±12.0% compared with the single administration of 11.0±4.3%. Conclusions: Our results suggest that the repeated transtracheal gene transfection using HVJ-liposome may have benefits for treatment of problems after lung transplantation. In addition, gene transfer using a flushing solution during harvest may provide an opportunity for gene manipulation in the setting of lung transplantation.
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