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Eur J Cardiothorac Surg 2005;27:768-773
© 2005 Elsevier Science NL

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

Department of Surgery (E1), Osaka University Graduate School of Medicine, Yamada-oka 2-2, Suita City, Osaka 565-0871, Japan

Received 20 June 2004; received in revised form 1 December 2004; accepted 23 December 2004.

^* Corresponding author. Address: Department of General Thoracic Surgery, Kure Medical Center, Kure National Hospital, Aoyamacho 3-1 Kure, Hiroshima 737-0023, Japan. Tel.: +81 823 22 3111; fax: +81 823 21 0478. (E-mail: omorik{at}kure-nh.go.jp).

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: ß-Galactosidase (ß-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 ß-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.

Key Words: Gene transfer • HVJ-liposome • Hybrid vector • Lung transplantation

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