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GENE THERAPY

Adeno-Associated Virus Type 6 (AAV6) Vectors Mediate Efficient Transduction of Airway Epithelial Cells in Mouse Lungs Compared to That of AAV2 Vectors

Christine L. Halbert, James M. Allen, A. Dusty Miller
Christine L. Halbert
Fred Hutchinson Cancer Research Center, Seattle, Washington 98109
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James M. Allen
Fred Hutchinson Cancer Research Center, Seattle, Washington 98109
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A. Dusty Miller
Fred Hutchinson Cancer Research Center, Seattle, Washington 98109
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DOI: 10.1128/JVI.75.14.6615-6624.2001
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ABSTRACT

Although vectors derived from adeno-associated virus type 2 (AAV2) promote gene transfer and expression in many somatic tissues, studies with animal models and cultured cells show that the apical surface of airway epithelia is resistant to transduction by AAV2 vectors. Approaches to increase transduction rates include increasing the amount of vector and perturbing the integrity of the epithelia. In this study, we explored the use of vectors based on AAV6 to increase transduction rates in airways. AAV vectors were made using combinations ofrep, cap, and packaged genomes from AAV2 or AAV6. The packaged genomes encoded human placental alkaline phosphatase and contained terminal repeat sequences from AAV2 or AAV6. We found that transduction efficiency was primarily dependent on the source of Cap protein, defined here as the vector pseudotype. The AAV6 and AAV2 pseudotype vectors exhibited different tropisms in tissue-cultured cells, and cell transduction by AAV6 vectors was not inhibited by heparin, nor did they compete for entry in a transduction assay, indicating that AAV6 and AAV2 capsid bind different receptors. In vivo analysis of vectors showed that AAV2 pseudotype vectors gave high transduction rates in alveolar cells but much lower rates in the airway epithelium. In contrast, the AAV6 pseudotype vectors exhibited much more efficient transduction of epithelial cells in large and small airways, showing up to 80% transduction in some airways. These results, combined with our previous results showing lower immunogenicity of AAV6 than of AAV2 vectors, indicate that AAV6 vectors may provide significant advantages over AAV2 for gene therapy of lung diseases like cystic fibrosis.

  • Copyright © 2001 American Society for Microbiology
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Adeno-Associated Virus Type 6 (AAV6) Vectors Mediate Efficient Transduction of Airway Epithelial Cells in Mouse Lungs Compared to That of AAV2 Vectors
Christine L. Halbert, James M. Allen, A. Dusty Miller
Journal of Virology Jul 2001, 75 (14) 6615-6624; DOI: 10.1128/JVI.75.14.6615-6624.2001

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Adeno-Associated Virus Type 6 (AAV6) Vectors Mediate Efficient Transduction of Airway Epithelial Cells in Mouse Lungs Compared to That of AAV2 Vectors
Christine L. Halbert, James M. Allen, A. Dusty Miller
Journal of Virology Jul 2001, 75 (14) 6615-6624; DOI: 10.1128/JVI.75.14.6615-6624.2001
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KEYWORDS

DNA-binding proteins
Dependovirus
Genetic Therapy
Transduction, Genetic

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