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Repeated delivery of adeno-associated virus vectors to the rabbit airway.

S E Beck¹, L A Jones, K Chesnut

¹Eudowood Division of Pediatric Respiratory Sciences, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA. sbeck@jhmi.edu

Journal of Virology

|October 9, 1999

Summary

Repeated delivery of adeno-associated virus (AAV) vectors for cystic fibrosis (CF) gene therapy is feasible in rabbits. Despite pre-existing antibodies, airway gene transfer remained effective and safe, suggesting serum antibody levels do not predict airway neutralization.

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Area of Science:

Gene Therapy
Respiratory Medicine
Virology

Background:

Recombinant adeno-associated virus (rAAV) vectors show efficient local expression in airways for CFTR correction.
Sustained correction of cystic fibrosis (CF) defects may require repeated rAAV vector administrations.
Previous studies demonstrated long-term expression after single rAAV-CFTR delivery in animal airways.

Purpose of the Study:

To assess the feasibility of repeated airway delivery of rAAV vectors in a rabbit model.
To investigate the impact of pre-existing anti-AAV antibodies on subsequent gene transfer efficiency.
To evaluate the safety and efficacy of repeated rAAV gene transfer to the airways.

Main Methods:

Rabbits received two doses of rAAV-CFTR, followed by a third dose of rAAV-GFP (serotype 2 or 3) via bronchoscopic delivery.

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Serum anti-AAV neutralizing antibody titers were measured before and after vector administration.

GFP expression and inflammatory responses were assessed in airway epithelial cells three weeks post-delivery.

Main Results:

High titers of serum anti-AAV neutralizing antibodies were generated after two doses.
GFP expression was observed in airway epithelial cells after the third dose, despite high antibody titers.
No significant difference in gene transfer efficiency or expression was found between AAV2 and AAV3 serotypes.
No significant inflammatory responses were observed following repeated vector exposure.

Conclusions:

Serum anti-AAV neutralizing antibody titers do not accurately predict in vivo airway neutralization.
Repeated airway delivery of rAAV vectors is safe and effective for gene transfer in the presence of pre-existing antibodies.
These findings support the potential for repeated rAAV-based gene therapy for respiratory diseases like CF.