MOLECULAR CONJUGATE-MEDIATED GENE-TRANSFER INTO ISOLATED HUMAN KIDNEYS

Advances in systemic immunosuppressive therapy for solid organ transpl antation have done little to decrease the percentage of allografts tha t eventually will develop chronic rejection, However, one of the promi ses of modern molecular biology includes the ability to introduce new genetic information into mammalian hosts. The ability to deliver genes and control their expression in the adult kidney has been described i n appropriate animal models, Consequently, gene transfer technology re presents a realistic therapeutic approach to modify the allogeneic kid ney before engraftment in an effort to decrease the incidence of postt ransplant dysfunction, To bridge the gap between animal studies and th e clinical application of this technology, we report the first genetic transfection of isolated human kidneys under conditions of organ pres ervation. Polymerase chain reaction, reversed transcription polymerase chain reaction, and in situ hybridization techniques demonstrated tha t an adenovirus-polylysine-deoxyribonucleic acid (DNA) complex can be used to insert a complementary DNA expression vector encoding beta-gal actosidase into the intact human kidney, Immunohistochemical and in si tu enzymatic analyses determined further that gene delivery and expres sion were localized in proximal tubular epithelial cells. Consequently , targeting of genes to perturb mediators of the local inflammatory re sponse may represent a rational therapeutic interventional strategy in chronic rejection of the kidney.