Achieving persistent expression is a prerequisite for effective genetic therapies for inherited disorders. These proof-of-concept studies focused on adeno-associated virus (AAV) administration to newborn monkeys. Serotype rh10 AAV expressing ovalbumin and green fluorescent protein (GFP) was administered intravenously at birth and compared with vehicle controls. At 4 months postnatal age, a second injection was administered intramuscularly, followed by vaccination at 1 year of age with ovalbumin and GFP. Ovalbumin was highest 2 weeks post administration in the treated monkey, which declined but remained detectable thereafter; controls demonstrated no expression. Long-term AAV genome copies were present in myocytes. At 4 weeks, neutralizing antibodies to rh10 were present in the experimental animal only. With AAV9 administration at 4 months, controls showed transient ovalbumin expression that disappeared with the development of strong anti-ovalbumin and anti-GFP antibodies. In contrast, increased and maintained ovalbumin expression was noted in the monkey administered AAV at birth, without antibody development. After vaccination, the experimental monkey maintained levels of ovalbumin without antibodies, whereas controls demonstrated high levels of antibodies. These preliminary studies suggest that newborn AAV administration expressing secreted and intracellular xenogenic proteins may result in persistent expression in muscle, and subsequent vector administration can result in augmented expression without humoral immune responses.
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This study was supported by the National Heart, Lung, and Blood Institute (NHLBI), National Institutes of Health (NIH) Center for Fetal Monkey Gene Transfer for Heart, Lung, and Blood Diseases (#HL85794; AFT); the Primate Center base operating grant (#OD011107) (AFT); NIH grants #NS071076, #NS071076-04S1, and #HD057555 (GSL); NIH grant AI108826-01 (GSL and AFT); and NIGMS Medical Genetics NIH T32 GM008243 and the Society of University Surgeons (DST). We thank Agustin Vega-Crespo and Sergio Duarte for their assistance with the fluorescence microscopy, Julie Johnston for helpful discussions and Roberto Calcedo for helpful discussions and for performing the neutralizing antibody assay.
The authors declare no conflict of interest.
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Tai, D., Hu, C., Lee, C. et al. Development of operational immunologic tolerance with neonatal gene transfer in nonhuman primates: preliminary studies. Gene Ther 22, 923–930 (2015). https://doi.org/10.1038/gt.2015.65
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