SUCCESSFUL GENE-TRANSFER TO THE PORCINE LIVER IN-VIVO WITH AN ADENOVIRAL VECTOR

In vivo gene transfer to the porcine liver was tested with adenoviral vector to achieve molecular biological graft modulation. In adult fema le pigs immunosuppressed with cyclophosphamide, cyclosporine, and pred nisolone, the liver was surgically isolated and flushed out with cold lactate Ringer solution (4 degrees C), by means of a pump-controlled b ypass of the portal vein and the inferior vena cava in Groups A and D, In Group A (n = 4), 2 x 10(11) pfu of the adenoviral vectors (pAdexCA LacZ) were injected through the left hepatic artery during cold ischem ia In Group B (n = 4), 2 x 1011 pfu of adenovirus vectors were injecte d through the auricular vein in a one-shot manner without a laparotomy , In Group C (n = 4), 2 X 10(11) pfu/ml of adenoviral vectors were inj ected through the hepatic artery in a one-shot manner, without a surgi cal isolation of the liver. Group D (n = 4) animals received the same protocol as Group A except for the fact that they did not receive the immunosuppressive regimen. In a polymerase chain reaction, a transfect ed LacZ sequence was detected until POD 28 in Group A but not in the o ther groups. In -bromo-4-chloro-3-indolyl-beta-D-palactopyranoside (X- gal) staining, only the Group A animals revealed apparent staining pre dominantly in the portal area at POD 2, which then continued to be rec ognized until POD 28. The in situ perfusion of the liver combined with immunosuppression is thought to provide an ideal environment for the Liver-directed adenovirus-mediated gene transfer to the porcine liver, by enabling a long contact with a high titer of the adenoviral vector (C) 1998 Academic Press.