Surgical organ perfusion method for gene transfer into cells of the perifollicular area of the spleen: an experimental trial on farm pigs

In an attempt to develop a gene therapy method for splenic and systemic dis eases, an evaluation was made of surgical methods for gene transfer into po rcine spleen. We have developed a continuous closed-circuit organ perfusion method for gene transfer into porcine spleen. For gene transfer, we used a type-5 replication defective adenovirus vector expressing the E. coli P-ga lactosidase gene as a reporter gene. In eight young 22-35 kg farm pigs, the spleen was perfused in vivo with the viral solution via laparotomy, for 30 or 60min. Gene transfer was determined visually on histological cryosectio ns after X-gal and PAS staining. Infusion of the viral solution through the splenic artery did not result in gene expression. Perfusion of spleen in v ivo resulted in P-galactosidase reporter gene expression in the macrophages located around capillaries terminating in the perifollicular zone and in t he red pulp examined after four days. The present results suggest that the surgically performed spleen perfusion method can be used for gene transfer in the treatment of diseases having splenic manifestations and in systemic diseases.