Surfactant protein A-producing cells in human fetal lung are good targets for recombinant adenovirus-mediated gene transfer.
Local delivery of Escherichia coli beta-galactosidase gene (beta-gal) to surfactant protein-A (SP-A)-producing cells by a replication-defective recombinant adenovirus (AdCMV.beta-gal) was tested in human 8-12-wk-old fetal lung explants cultured in Waymouth's medium. In contrast to uninfected explants, direct addition of 0.8-1.6 x 10(6) plaque-forming units of AdCMV.beta-gal resulted in beta-galactosidase (beta-Gal)-specific staining of the pulmonary epithelium. SP-A localization by indirect immunofluorescence showed positive specific staining of the beta-Gal+ lung epithelial cells, demonstrating that recombinant-defective adenoviruses efficiently transfer reporter genes to fetal lung SP-A+ cells. The reporter gene expression in SPA+ cells persisted for more than 1 mo. No apparent alteration of morphology, phenotype, and growth was observed. The in vitro human lung model described may be useful for testing DNA constructs for vector-mediated gene therapy, as an approach to the treatment of congenital defects and neonatal disorders, such as respiratory distress syndrome and bronchopulmonary dysplasia.
Messina
E
E
Experimental Center on Gene Therapy and Diagnosis, Gabriele D'Annunzio University, Chieti, Italy.
Muhlhauser
J
J
Giuliano
M
M
Pandolfi
A
A
Morgese
G
G
Procopio
A
A
D000260:Adenoviruses, Human / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005069:Evaluation Studies as Topic; D005315:Fetal Diseases / Q000628:therapy; D005326:Fetal Proteins / Q000096:biosynthesis*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007231:Infant, Newborn; D007763:Lac Operon; D008168:Lung / Q000166:cytology / Q000196:embryology / Q000378:metabolism*; D011510:Proteolipids / Q000096:biosynthesis*; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000096:biosynthesis*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014779:Virus Replication
D005326:Fetal Proteins; D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D011993:Recombinant Fusion Proteins