Adenovirus-mediated gene transfer using in-situ perfusion of the liver graft.
To establish an efficient technique for adenovirus-mediated gene transfer in liver transplantation, we evaluated the in situ perfusion of liver grafts. The grafts were perfused in situ with 1 x 10(10) of E1-deleted, replication-defective adenoviral vectors encoding the LacZ gene driven by the human CMV promoter, either through the hepatic artery (group 1) or the portal vein (group 2). Group 3 animals served as negative controls; their liver grafts were perfused with lactated Ringer's solution through the portal vein. PCR confirmed the presence of viral DNA in every graft perfused with viral vectors. In X-gal staining, positive staining was observed almost exclusively at the portal triad in group 1, whereas in group 2 minimal staining was observed, predominantly in the parenchymal area. Protein production from the transfected gene was confirmed by a functional protein assay; the values were 0.16% +/- 0.07% liver protein in group 1, 0.13% +/- 0.02% in group 2, and 0.007% +/- 0.0003% in group 3 on postoperative day 2. In conclusion, in situ perfusion of the viral vectors through the hepatic artery resulted in an effective expression of the transfected gene, predominantly at the portal triad.
Shiraishi
M
M
First Department of Surgery, University of Ryukyu, School of Medicine, Okinawa, Japan.
Kusano
T
T
Hara
J
J
Hiroyasu
S
S
Shao-ping
M
M
Makino
Y
Y
Muto
Y
Y
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D005822:Genetic Vectors; D006085:Graft Survival; D006499:Hepatic Artery; D008102:Liver Circulation; D016031:Liver Transplantation / Q000379:methods*; D010477:Perfusion; D011169:Portal Vein; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection / Q000379:methods*