ADENOVIRUS-MEDIATED GENE-TRANSFER USING IN-SITU PERFUSION OF THE LIVER GRAFT

To establish an efficient technique for adenovirus-mediated gene trans fer in liver transplantation, we evaluated the in situ perfusion of li ver grafts. The grafts were perfused in situ with 1 x 10(10) of E1-del eted, replication-defective adenoviral vectors encoding the LacZ gene driven by the human CMV promoter. either through the hepatic artery (g roup 1) or the portal vein (group 2). Group 3 animals served as negati ve controls; their liver grafts were perfused with lactated Ringer's s olution through the portal vein. PCR confirmed the presence of viral D NA in every graft perfused with viral vectors. In X-gal staining, posi tive staining was observed almost exclusively at the portal tried in g roup 1: whereas in group 2 minimal staining was observed, predominantl y in the parenchymal area, Protein production from the transfected gen e 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 re sulted in an effective expression of the transfected gene, predominant ly at the portal triad.