In vivo model of adeno-associated virus vector persistence and rescue

Afione, S A; Conrad, C K; Kearns, W G; Chunduru, S; Adams, R; Reynolds, T C; Guggino, W B; Cutting, G R; Carter, B J; Flotte, T R

doi:10.1128/jvi.70.5.3235-3241.1996

Cited by 223Virus-based gene therapy researchRNA Interference and Gene DeliveryNeurogenetic and Muscular Disorders Research

In vivo model of adeno-associated virus vector persistence and rescue

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Afione, S A, Conrad, C K, Kearns, W G, Chunduru, S, Adams, R, Reynolds, T C, Guggino, W B, Cutting, G R +2 more·1996·Journal of Virology

DOI 10.1128/jvi.70.5.3235-3241.1996PMC PMC190187

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Abstract

Gene therapy vectors based on human DNA viruses could be mobilized or rescued from individuals who are subsequently infected with the corresponding wild-type (wt) helper viruses. This phenomenon has been effectively modeled in vitro with both adenovirus (Ad) and adeno-associated virus (AAV) vectors but has not previously been studied in vivo. In the current study, we have developed an in vivo model to study the interactions of a recombinant AAV vector (AAV-CFTR) with wt AAV type 2 (AAV2) and a host range mutant Ad (Ad2HR405) for which monkey cells are permissive (D.E.Brough, S.A.Rice, S.Sell, and D.F.Klessig, J. Virol. 55:206-212, 1985). AAV-CFTR was administered to the respiratory epithelium of the nose or lung of rhesus macaques. Primary cells were harvested from the infusion site at time points up to 3 months after vector administration to confirm vector DNA persistence. Vector DNA was present in episomal form and could be rescued in vitro only by addition of wt AAV2 and Ad. In in vivo rescue studies, vector was administered before or after wt-AAV2 and Ad2HR405 infection, and the shedding of AAV-CFTR was examined. Ad2HR405 and wt-AAV2 infections were established in the nose with concomitant administration. wt-AAV2 replication occurred in the lung when virus was administered directly at a high titer to the lower respiratory tract. AAV-CFTR vector rescue was also observed in the latter setting. Although these studies were performed with small numbers of animals within each group, it appears that AAV-CFTR DNA persists in the primate respiratory tract and that this model may be useful for studies of recombinant AAV vector rescue.

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Authors: Afione, S. A., Conrad, C. K., Kearns, W. G., Chunduru, S., Adams, R., Reynolds, T. C., Guggino, W. B., Cutting, G. R., Carter, B. J., & Flotte, T. R.
Affiliations: Johns Hopkins University (US)
ORCID: Conrad, C K
Kearns, W G
Adams, R
Reynolds, T C
Cutting, G R
Flotte, T R
Venue: Journal of Virology
Publisher: American Society for Microbiology
Published: 1996-05
Volume: 70
Issue: 5
Pages: 3235-3241
ISSN: 0022-538X (ISSN-L)
1098-5514
DOI: 10.1128/jvi.70.5.3235-3241.1996
PMC: PMC190187
OpenAlex: W2159706578
Cited by: 223
References: 26
Citations by year: 2026: 1 · 2025: 2 · 2024: 4 · 2022: 8 · 2021: 4 · 2020: 10 · 2019: 3 · 2018: 4 · 2017: 7 · 2016: 3 · 2015: 2 · 2014: 4 · 2013: 6 · 2012: 4
FWCI: 11.20
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Journal of Virology (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: National Heart, Lung, and Blood Institute (P01HL051811, HL 51811.)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Health Sciences › Medicine › Genetics › Neurogenetic and Muscular Disorders Research
Keywords: adeno-associated virus vectors, vector persistence, AAV2, adenovirus type 2, rhesus monkeys, respiratory tract, gene therapy vectors
MeSH terms: Dependovirus, Adenoviridae, Animals, Base Sequence, Bronchi, Bronchoalveolar Lavage Fluid, Cell Line, Cells, Cultured, Epithelial Cells, Epithelium, Genetic Vectors, HeLa Cells, Helper Viruses, Humans, Macaca mulatta, Molecular Sequence Data, Recombination, Genetic, Genetic Therapy, Polymerase Chain Reaction, Mutagenesis, Virus Shedding, In Situ Hybridization, Fluorescence, DNA Primers, Cystic Fibrosis Transmembrane Conductance Regulator

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