Induction of immunity to antigens expressed by recombinant adeno-associated virus depends on the route of administration.
Recombinant adeno-associated virus (rAAV) is a replication-defective parvovirus which is being explored as a vector for gene therapy because of its broad host range, excellent safety profile, and durable transgene expression in infected hosts. rAAV has also been reported by several groups to induce little or no immune response to its encoded transgene products. In this study we examined the immunogenicity of rAAV by studying the immune response of C57BL/6 mice to a single dose of rAAV-encoding ovalbumin (AAV-Ova) administered by a variety of routes. Mice injected with AAV-Ova intraperitoneally (ip), intravenously, or subcutaneously developed potent ovalbumin-specific cytotoxic T lymphocytes (CTL) as well as anti-ovalbumin antibodies and antibodies to AAV. In contrast, mice injected with AAV-Ova intramuscularly developed a humoral response to the virus and the transgene but minimal ovalbumin-specific CTLs. The induced CTL response after ip administration of AAV-Ova protected mice against a subsequent tumor challenge with an ovalbumin-transfected B16 melanoma cell line. Studies of the mechanism by which AAV-Ova induces CTL confirmed that the virus delivers the transgene product into the classical MHC class I pathway of antigen processing. Mice that previously had been exposed to rAAV vectors failed to develop ovalbumin-specific CTL following administration of AAV-Ova. Analysis of these mice revealed the presence of circulating anti-AAV antibodies that blocked rAAV transduction in vitro and inhibited CTL induction in vivo. These results suggest a possible role for rAAV in the immunotherapy of malignancies and viral infections, although induced antibody responses to AAV may limit its ability to be administered for repeated vaccinations.
Department of Pathology, Stanford University School of Medicine, Stanford, California 94305, USA.
D G
DG
Brockstedt
G M
GM
Podsakoff
L
L
Fong
G
G
Kurtzman
W
W
Mueller-Ruchholtz
E G
EG
Engleman
D000818:Animals; D000917:Antibody Formation; D000956:Antigens, Viral / Q000008:administration & dosage*; D000229:Dependovirus / Q000276:immunology*; D004333:Drug Administration Routes; D005260:Female; D007111:Immunity, Cellular / Q000276:immunology; D007273:Injections, Intramuscular; D051379:Mice; D008810:Mice, Inbred C57BL; D010047:Ovalbumin / Q000276:immunology; D011994:Recombinant Proteins / Q000008:administration & dosage
D000956:Antigens, Viral; D011994:Recombinant Proteins; D010047:Ovalbumin