Adenovirus-mediated transfer of beta-galactosidase and prourokinase genes into vein grafts

Objective To study the feasibility of adenovirus mediated gene transfer int o vein grafts and the role of the prourokinase gene in protecting vein graf ts from thrombosis. Methods Fifty-two Wistar rats underwent implantation of reversed autologous jugular vein interposition grafts in the common carotid arteries. Jugular veins were excised and distended with solution containing three different a denovirus vectors (AdV(5)-CMV, group I; Adv(5)-CMV/LacZ, group II; Adv(5)-C MV/Pro-UK, group Ill) for 30 min, then the jugular veins were reversed and interposed into the divided carotid arteries, and end-to-end anastomoses we re performed. The amount of Cr-51-labeled platelets in vein grafts of group I and group III was counted 24 hours postoperatively. On the 14th day, the vein grafts were harvested to examine beta-galactosidase activity and prou rokinase (Pro-UK) activity and observe thrombosis in vein grafts. reversed arteries, Results Extensive blue coloration in the area of intima and media of each v ein graft in group II was observed. No blue coloration was seen in group I. Pro-UK activity was not detected in the vein grafts of group I. In group I II, the amount of Pro-UK gene expression was 308 IU/g tissue. The amount of Cr-51-labeled platelets in group I and group II was (123.7 +/- 19.4) x 10( 6)/g dry wt, (34.4 +/- 5.3) x 10(6)/g dry wt, respectively. The thrombosis rate and occlusion rate of the vein grafts in group I were 30% and 10%, res pectively. In group III, all vein grafts were patent and free of thrombosis . Conclusions Ex vivo gene transfer before vein grafting is feasible using re plication deficient recombinant adenovirus and results in a high level of g ene expression in vivo. Direct transfer of the Pro-UK gene into vein grafts may prevent thrombosis.