Two novel adeno-associated viruses from cynomolgus monkey: pseudotyping characterization of capsid protein.
We demonstrated the presence of two adeno-associated viruses (AAVs), designated AAV10 and AAV11, in cynomolgus monkeys by isolating and sequencing the entire viral coding regions from the monkey DNA. AAV10 and AAV11 capsid proteins shared 84% and 65%, respectively, of amino acids with AAV2. A phylogenetic analysis of AAV capsid proteins showed that AAV10 and AAV11 resembled most AAV8 and AAV4, respectively. To characterize the capsid protein, we pseudotyped an AAV2 vector with the monkey AAV capsid proteins and examined the resulting pseudotypes AAV2/10 and AAV2/11, in comparison with the AAV2 vector, for their host ranges in cell lines and tissue tropism in mice. AAV2/10 and AAV2/11 transduced primate cells less efficiently than AAV2. Whereas AAV2 transduced undifferentiated C2C12 mouse myoblasts more efficiently than differentiated ones, AAV2/10 and AAV2/11 transduced the undifferentiated myoblasts less efficiently than differentiated ones. Three weeks after injection to the muscle of the hind legs, AAV2/10 and AAV2 induced transgene expression similarly, but AAV2/11 did not transduce the skeletal muscle. Six weeks after systemic administration, transduced vector DNA was detected by PCR in the liver and spleen of mice inoculated with AAV2, in the liver, heart, muscle, lung, kidney, and uterus of mice with AAV2/10, and the muscle, kidney, spleen, lung, heart, and stomach of mice with AAV2/11. Mouse antisera against capsid protein VP2 of the three AAVs neutralized the respective vector particles in a type-specific manner. The results indicate that AAV10 and AAV11 capsid proteins, which are antigenically distinct from each other and AAV2, are likely to determine their host ranges and tissue tropism that are different from AAV2s, suggesting that cynomolgus AAVs could provide a broader choice of pseudotype AAV vectors for gene therapy.
Mori
Seiichiro
S
Division of Molecular Genetics, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan.
Wang
Lina
L
Takeuchi
Takamasa
T
Kanda
Tadahito
T
D000595:Amino Acid Sequence; D000818:Animals; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D002460:Cell Line; D004279:DNA, Viral / Q000737:chemistry / Q000302:isolation & purification; D000229:Dependovirus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D005260:Female; D005822:Genetic Vectors; D006321:Heart / Q000821:virology; D007668:Kidney / Q000821:virology; D008099:Liver / Q000821:virology; D008168:Lung / Q000821:virology; D008252:Macaca fascicularis / Q000821:virology*; D051379:Mice; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000821:virology; D032446:Myoblasts / Q000166:cytology / Q000821:virology; D009500:Neutralization Tests; D010802:Phylogeny; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000821:virology; D013270:Stomach / Q000821:virology; D014161:Transduction, Genetic
D036022:Capsid Proteins; D004279:DNA, Viral; C442553:VP2 protein, Dependovirus