1. ¹INSERM UMR 649, CHU Nantes, Faculté de Médecine, Université de Nantes, Nantes, France
2. ²INRA UMR 703, Ecole Nationale Vétérinaire, Nantes, France
3. ³EFS Pays de la Loire, Nantes, France
4. ⁴Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida, USA
5. ⁵School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
6. ⁶Service des Urgences, Ecole Nationale Vétérinaire, Nantes, France
7. ⁷Service de Chirurgie Pédiatrique, CHU Hôtel-Dieu, Nantes, France
8. ⁸The Children's Hospital of Philadelphia, Department of Pediatrics, University of Pennsylvania, Philadelphia, Pennsylvania, USA
9. ⁹Howard Hughes Medical Institute, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA
10. ¹⁰Department of Surgery, University of Pennsylvania, Philadelphia, Pennsylvania, USA
11. ¹¹INSERM UMR 643, CHU Nantes, Institut de Transplantation et de Recherche en Transplantation, Faculté de Médecine, Université de Nantes, Nantes, France

Correspondence: Caroline Le Guiner, INSERM UMR 649, Bâtiment Jean Monnet, CHU Hôtel-Dieu, 30 Boulevard Jean Monnet, 44035 Nantes Cedex 01, France. E-mail: caroline.le-guiner@univ-nantes.fr

Received 31 January 2008; Accepted 2 April 2008; Published online 6 May 2008.

Abstract

We developed a drug-free regional intravenous (RI) delivery protocol of recombinant adeno-associated virus (rAAV) 1 and 8 to an entire limb in the nonhuman primate (NHP), and compared the results with those produced by intramuscular (IM) delivery of the same dose of vector. We show that RI delivery of both serotypes was remarkably well tolerated with no adverse side-effects. After IM, muscle transduction was restricted to the site of injection with a high number of vector copies per cell for rAAV1. In contrast, although RI delivery resulted in a lower vector copy per cell, it was detectable in the vast majority of muscles of the injected limb. The amounts of circulating infectious rAAV were similar for both serotypes and modes of delivery. At autopsy at up to 34 months after vector administration, similar biodistribution patterns were found for both vectors and for both modes of delivery, with numerous organs found to be positive for vector sequence when assayed using PCR and Southern blot. Altogether, we demonstrated that RI is a simple and efficient transduction protocol in NHPs, resulting in higher expression of the transgene with a lower number of vector genomes per cell. However, regardless of the mode of delivery, concerns continue to be raised by the presence of vector sequences detected at distant sites.