RECOMBINANT CFTR DETECTION IN CF TRACHEAL EPITHELIAL-CELLS FOLLOWING IN-VITRO LIPOSOME-MEDIATED GENE-TRANSFER

The efficacy of CFTR gene transfer mediated by cationic liposomes Dc-C hol/DOPE into cystic fibrosis (CF) tracheal epithelial cells carrying defective processing mutations (S549N/N1303K), was assessed by studyin g mRNA and protein expression of the recombinant product. Appreciable levels of mRNA transcripts were detected 48 h after transfection, whil e complete translocation of the recombinant CFTR to the apical membran e of epithelial cells was observed after 72 h following transfection. Our results suggest that in vitro restoration of a normal CFTR process ing and migration to the cell plasmalemma requires 72 h at least as de monstrated by immunocyto-fluorescence using the monoclonal antibody MA TG 1016. These findings are relevant onto gene transfer phase I clinic al studies.