Application of HVJ-liposome mediated gene transfer in lung transplantation-distribution and transfection efficiency in the lung.
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
Beta-galactosidase (beta-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 microg of the beta-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.
Ohmori
Kenichi
K
Department of Surgery (E1), Osaka University Graduate School of Medicine, Yamada-oka 2-2, Suita City, Osaka 565-0871, Japan. omorik@kure-nh.go.jp <omorik@kure-nh.go.jp>
Takeda
Shin-ichi
S
Miyoshi
Shinichiro
S
Minami
Masato
M
Nakane
Shigeru
S
Ohta
Mitsunori
M
Sawa
Yoshiki
Y
Matsuda
Hikaru
H
D000818:Animals; D015870:Gene Expression; D005822:Genetic Vectors / Q000008:administration & dosage*; D007267:Injections; D007269:Injections, Intra-Arterial; D008081:Liposomes; D008168:Lung / Q000201:enzymology*; D016040:Lung Transplantation / Q000379:methods*; D010957:Plasmids; D011651:Pulmonary Artery; D051381:Rats; D017207:Rats, Sprague-Dawley; D029082:Sendai virus / Q000235:genetics*; D014132:Trachea; D014161:Transduction, Genetic / Q000379:methods*; D016896:Treatment Outcome; D001616:beta-Galactosidase / Q000235:genetics*
D008081:Liposomes; D001616:beta-Galactosidase