Adenovirus-Augmented, Receptor-Mediated Gene Delivery and Some Solutions to the Common Toxicity Problems
Several years ago it was reported that the entry of adenovirus particles could augment the uptake of polylysine-condensed DNA molecules (Curiel et al. 1991), a phenomenon similar to previously described adenovirus augmentation of both fluid phase markers and receptor-bound molecules (Fernández-Puentes and Carrasco 1980; Fitzgerald et al. 1983). This use of adenovirus to enhance gene delivery has proven to be a fruitful approach. A number of reviews of the general properties of this system have recently been published (Curiel 1993; Cotten and Wagner 1993; Wagner et al. 1994). In this review I will provide a summary of the current methods of generating adenovirus-DNA transfection complexes as well as discussing two of the toxicity problems that we have encountered with this system and describing some solutions to these problems. The first problem encountered was due to virus gene expression and leakiness in the replication defect of the commonly used defective adenovirus strains. This problem was solved by developing a psoralen inactivation method that inactivates the virus DNA and blocks viral gene expression and replication without impairing the entry functions of the virus (Cotten et al. 1994b). The second problem was due to a toxicity generated by cytoplasmic delivery of lipopolysaccharide (LPS, endotoxin) that frequently contaminates bacterial DNA preparations. The LPS problem has been solved by identifying reliable methods of removing LPS from DNA (Cotten et al. 1994a) or by including LPS-binding polymyxin in the transfection medium.
KeywordsGene Delivery Viral Gene Expression Transfection Complex Entry Function Penton Base
Unable to display preview. Download preview PDF.
- Buschle M, Cotten M, Kirlappos H, Mechtler K, Schaffner G, Zauner W, Bimstiel ML, Wagner E (1995) Receptor-mediated gene transfer into human T-lymphocytes via binding of DNA/CD3 antibody particles to the CD3 T cell receptor complex. Human Gene Therapy (in press)Google Scholar
- Cotten M, Wagner E, Zatloukal K, Phillips S, Curiel DT, Bimstiel ML, (1992) High-efficiency receptor-mediated delivery of small and large (48kb) gene constructs using the endosome disruption activity of defective or chemically inactivated adenovirus particles. Proc Natl Acad Sci USA 89: 6094–6098PubMedCrossRefGoogle Scholar
- Hanson CV (1992) Photochemical inactivation of viruses with psoralens: an overview. Blood Celisi 8: 7–25Google Scholar
- Wagner E, Zatloukal K, Cotten M, Kirlappos H, Mechtler K, Curiel D, Bimstiel ML (1992a) Coupling of adenovirus to transferrin-polylysine/DNA complexes greatly enhances receptor-mediated gene delivery and expression of transfected cells. Proc Natl Acad Sci USA 89: 6099–6103PubMedCrossRefGoogle Scholar
- Wagner E, Plank C, Zatloukal K, Cotten M, Bimstiel ML (1992b) Influenza virus hemagglutinin HA-2 N-terminal fusogenic peptides augment gene transfer by transferrin-polylysine-DNA complexes: toward a synthetic virus-like gene-transfer vehicle. Proc Natl Acad Sci USA 89: 7934–7938PubMedCrossRefGoogle Scholar
- White E, Gooding LR (1994) Regulation of apoptosis by human adenoviruses. (Apoptosis II: the molecular basis of apoptosis in disease.) Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
- Yoshinaga S, Dean N, Han M, Berk AJ (1986) Adenovirus stimulation of transcription by RNA polymerase III: evidence for an E1A-dependent increase in transcription factor IIIC concentration. EM BO J 5: 343–354Google Scholar
- Zatloukal K, Cotten M, Berger M, Schmidt W, Wagner E, Bimstiel ML (1994) In vivo production of human factor VIII in mice after intrasplenic implantation of primary fibroblasts transfected by receptor-mediated adenovirus-augmented gene delivery. Proc Natl Acad Sci USA 91: 5148–5152PubMedCrossRefGoogle Scholar