Expression of the hepatitis C virus genome in rat liver after cationic liposome-mediated in vivo gene transfer.
The lack of a small animal model of hepatitis C virus (HCV) infection has impeded elucidation of the pathogenesis of HCV. The aim of this study was to develop an HCV-expressing animal model by means of cationic liposome-mediated in vivo gene transfer. To examine the feasibility of this strategy, pActLacZ, an expression vector composed of the LacZ gene driven by the beta-actin promoter, complexed with lipofectin, was injected retrogradely into the common bile ducts of adult rats. X-Gal histochemical staining clearly showed that the LacZ gene was expressed in hepatocytes, but not in biliary epithelial cells. Maximal expression was observed at a DNA to lipofectin ratio of 1:4. Based on this observation, pAGS3M091, an expression vector containing the full length of HCV complementary DNA (cDNA) preceded by the beta-actin promoter, was evaluated. Two days after in vivo intrabiliary administration of pAGS3M091 complexed with lipofectin, polymerase chain reaction (PCR) amplification of reverse-transcribed liver RNA demonstrated the 5' and 3' portions of HCV transcripts derived from pAGS3M091. Immunohistochemical analysis showed the HCV core protein in a small number of hepatocytes scattered in the hepatic lobules. We conclude that the full-length HCV genome was successfully expressed in adult rat liver by means of cationic liposome-mediated in vivo gene transfer. This model will be useful for determining the immunopathological role of HCV in vivo.
Takehara
T
T
First Department of Medicine, Osaka University Medical School, Japan.
Hayashi
N
N
Miyamoto
Y
Y
Yamamoto
M
M
Mita
E
E
Fusamoto
H
H
Kamada
T
T
D000818:Animals; D002412:Cations; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics*; D007763:Lac Operon; D008081:Liposomes; D008099:Liver / Q000502:physiology*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D014158:Transcription, Genetic; D014162:Transfection; D014758:Viral Core Proteins / Q000378:metabolism
D002412:Cations; D018076:DNA, Complementary; D008081:Liposomes; D014758:Viral Core Proteins