Adenovirus-Mediated Transfer of a Recombinant α1-Antitrypsin Gene to the Lung Epithelium in Vivo

Abstract

The respiratory epithelium is a potential site for somatic gene therapy for the common hereditary disorders alpha 1-antitrypsin (alpha 1AT) deficiency and cystic fibrosis. A replication-deficient adenoviral vector (Ad-alpha 1AT) containing an adenovirus major late promoter and a recombinant human alpha 1AT gene was used to infect epithelial cells of the cotton rat respiratory tract in vitro and in vivo. Freshly isolated tracheobronchial epithelial cells infected with Ad-alpha 1AT contained human alpha 1AT messenger RNA transcripts and synthesized and secreted human alpha 1AT. After in vivo intratracheal administration of Ad-alpha 1AT to these rats, human alpha 1AT messenger RNA was observed in the respiratory epithelium, human alpha 1AT was synthesized and secreted by lung tissue, and human alpha 1AT was detected in the epithelial lining fluid for at least 1 week.

Stepwise dismantling of adenovirus 2 during entry into cells

“…Besides being invaluable as model viruses in molecular biology, they are important as pathogens and as transforming viruses (see Grunhaus and Horwitz, 1992;Horwitz, 1990). Recently, they have acquired special interest as one of the most effective viral vehicles for gene therapy (Gotten et al, 1993;Jaffe et al, 1992;Ragot et al, 1993;Rosenfeld et al, 1991Rosenfeld et al, , 1992. Advantages of this virus for gene delivery are the abilities to infect nondividing cells, to enter a large number of cell types, and to be readily available in purified form as stable, recombinant, and replication-defective particles that can carry large inserts (see Mulligan, 1993).…”

Adenoviral reporter gene transfer to the human trabecular meshwork does not alter aqueous humor outflow. Relevance for potential gene therapy of glaucoma

“…Adenoviral vectors have proven to be very effective in transferring genes to nondividing cells [10][11][12][13] and for this reason have became the vectors of choice for gene transfer to the eye. [14][15][16][17][18][19] Previous experiments from our laboratory have shown that a single injection into the anterior chamber of rabbits resulted in effective gene transfer into the cells of the trabecular meshwork.…”

Induction of therapeutic antitumor antiangiogenesis by intratumoral injection of genetically engineered endostatin-producing Semliki Forest virus

“…Furthermore, although adenovirus vectors can express high levels of foreign genes, these systems are more complex than SFV and express many highly antigenic virus -specific gene products, including structural proteins. 32 In contrast, the current SFV vector expresses only the four viral replicase proteins (ns P 1 -4 ) required for RNA amplification in the infected cells. This self -replicating RNA was shown to produce more than 200% the amount of antigen produced using a conventional DNA immunogen.…”