Persistent gene expression in rat liver in vivo by repetitive transfections using HVJ-liposome.
Most viral vectors are highly immunogenic and are of limited use for somatic gene therapy that requires repetitive administrations. We have developed a highly efficient gene transduction procedure useful for repetitive transfections using liposome containing hemagglutinating virus of Japan (HVJ-liposome). The Escherichia coli beta-galactosidase (beta-gal) gene was embodied in HVJ-liposome, and introduced directly into the caudal lobe of rat liver that was transiently isolated from a systemic circulation. A 116 kDa beta-gal protein was detected in transfected rat liver tissues by Western blot analysis and it was expressed in more than two-thirds of the liver by histological staining. It was found that the transfection efficiency was not affected by repetitive transfections. In support of these findings, antibody response to HVJ-liposome detected in the rat sera was weak and transient. Furthermore, cytotoxic T lymphocytes were not elicited against autologous rat hepatocytes that were transfected in vivo using HVJ-liposome. Thus, our results demonstrate that the isolation of a target liver from systemic circulation and the direct administration of foreign genes using HVJ-liposomes are useful for high gene transduction and persistent gene expression in the liver.
Hirano
T
T
First Department of Surgery, Hyogo College of Medicine, Nishinomiya, Japan.
Fujimoto
J
J
Ueki
T
T
Yamamoto
H
H
Iwasaki
T
T
Morisita
R
R
Sawa
Y
Y
Kaneda
Y
Y
Takahashi
H
H
Okamoto
E
E
D000818:Animals; D015153:Blotting, Western; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000008:administration & dosage*; D007124:Immunoenzyme Techniques; D008081:Liposomes; D008099:Liver / Q000276:immunology / Q000378:metabolism*; D008297:Male; D016133:Polymerase Chain Reaction; D051381:Rats; D017208:Rats, Wistar; D018108:Respirovirus; D013602:T-Lymphocytes, Cytotoxic; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics*
D008081:Liposomes; D001616:beta-Galactosidase