Isolation and evaluation of novel adeno-associated virus sequences from porcine tissues.
High antigenic compatibility and low toxicity is associated with xenograft transplantation of porcine tissues in immunodeficient human recipients. We hypothesized that adeno-associated viruses (AAVs) of porcine origin could be highly compatible to human tissues and thus of good efficiency and low toxicity for in vivo gene transfer. Porcine tissues were screened by PCR for the presence of AAV using primers designed to bind conserved regions and amplify variable regions of an alignment of several AAV sequences available on GenBank. We isolated new AAV capsid sequences from porcine tissues and successfully generated a recombinant AAV2/po1 vector by transfection. The AAV2/po1 vector was not cross-neutralized by antisera generated against all other commonly used AAVs (serotype 1, 2, 3, 4, 5, 7 and 8) indicating a distinct antigenic profile. Preexisting immunity to AAVpo1 could not be detected in the human sera evaluated. In mice, AAV2/po1 particles expressing beta-galactosidase or green fluorescent protein demonstrated high transduction efficiency in muscle fibers and the retina after intramuscular or intraocular administration. Biodistribution experiments following systemic administration showed efficient gene transfer exclusively in muscle fibers. Novel AAVs derived from porcine tissues may contribute to the generation of new preventive or curative clinical modalities acceptable for human use.
Bello
A
A
Special Pathogens Program, Public Health Agency of Canada, National Microbiology Laboratory, Manitoba, Canada.
Tran
K
K
Chand
A
A
Doria
M
M
Allocca
M
M
Hildinger
M
M
Beniac
D
D
Kranendonk
C
C
Auricchio
A
A
Kobinger
G P
GP
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D004279:DNA, Viral / Q000302:isolation & purification; D000229:Dependovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D005822:Genetic Vectors / Q000008:administration & dosage / Q000493:pharmacokinetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D012160:Retina / Q000378:metabolism; D016415:Sequence Alignment; D034421:Sus scrofa / Q000821:virology*; D014161:Transduction, Genetic; D056189:Viral Tropism
D004279:DNA, Viral