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Application of HVJ-liposome mediated gene transfer in lung transplantation—distribution and transfection efficiency in the lung

Application of HVJ-liposome mediated gene transfer in lung transplantation—distribution and transfection efficiency in the lung,Kenichi Ohmori,Shin-ic

Application of HVJ-liposome mediated gene transfer in lung transplantation—distribution and transfection efficiency in the lung  
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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: b-Galactosidase (b-gal) plasmid DNA was introduced into lung tissues using the HVJ-liposome method. Two groups of Sprague- Dawley (SD) rats received intratracheal instillation of 10 mg of the b-gal gene, once on Day 0 in 1 group (Group Tb-1, nZ4) and 3 times on Days 0-2 in another (Group Tb-3, nZ4). In a third group of SD rats (Group Tx, nZ5), an orthotopic left lung transplantation was performed after the donor lung was flushed with an HVJ-liposome complex solution and preserved for 1 h. 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 (0.2G0.1 and 4.0G1.8%), respectively, but a moderate degree of transfection to the airway (11.0G7.1 and 28.0G20.7%) and alveolar cells (3.0G1.8 and 6.0G3.6%). Three repetitive injections via the airway increased gene expression in airway epithelial cells of 41.0G12.0% compared with the single administration of 11.0G4.3%. 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. q 2005 Elsevier B.V. All rights reserved.
Published in 2010.
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