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Intravenous administration of an E1/E3-deleted adenoviral vector induces tolerance to factor IX in C57BL/6 mice

Gene Therapy volume 8, pages 354–361 (2001)Cite this article

Abstract

Inbred immunocompetent C57BL/6 mice have been a favored strain to study transgene expression of human blood coagulation factor IX (hF.IX) from viral vectors because systemic expression of the secreted protein is not limited by antibody responses following intravenous (i.v.) injection of vector. For example, i.v. injection of an adenoviral (Ad) vector results in sustained expression of hF.IX in normal or hemophilic C57BL/6 mice, while anti-hF.IX antibodies rapidly emerge in other strains (Gene Therapy 4: 473; Blood 91: 784). To investigate these observations further, we injected naive C57BL/6 mice and C57BL/6 mice with pre-existing anti-hF.IX with Ad-hF.IX vector via peripheral vein. All mice expressed hF.IX antigen without detectable anti-hF.IX, even when challenged with hF.IX in different immunogenic settings at later time points. Moreover, in mice with pre-existing immunity, anti-hF.IX titers diminished to undetectable levels after i.v. administration of Ad-hF.IX. Lymphocytes from mice that had received Ad-hF.IX i.v. failed to proliferate when stimulated with hF.IX in vitro after the animals had been repeatedly challenged with hF.IX protein formulated in complete Freund's adjuvant. Thus, absence of anti-hF.IX in C57BL/6 mice after i.v. injection of Ad vector is not due to ignorance to the foreign transgene product. Similar experiments in other strains showed that immune tolerance to hF.IX does not correlate with the strain haplotype or expression of IL-10 cytokine. Given the well-documented immunogenicity of the first-generation adenoviral vector, data from C57BL/6 mice may therefore grossly underestimate immunological consequences in certain gene therapy protocols.

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Acknowledgements

This work was supported by NIH grant R01 HL61921 to KAH and NIH grant AI-27828 to JPF. PAF was supported by grants from the Medical Research Council (MRC, UK) and the Katherine Dormandy Trust for Hemophilia. RWH is supported by a Career Development Award from the National Hemophilia Foundation. The authors thank Dr Robert Coffman for providing the BALB/c IL-10 knock-out mice and Avigen, Inc., a company in which KAH holds equity, for providing AAV vector.

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Authors and Affiliations

  1. Departments of Pediatrics and Pathology, University of Pennsylvania Medical Center and The Children's Hospital of Philadelphia, Philadelphia, PA, USA

    PA Fields, E Armstrong, JN Hagstrom, VR Arruda, KA High & RW Herzog

  2. School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA

    ML Murphy & JP Farrell

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  1. PA Fields
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  2. E Armstrong
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Fields, P., Armstrong, E., Hagstrom, J. et al. Intravenous administration of an E1/E3-deleted adenoviral vector induces tolerance to factor IX in C57BL/6 mice. Gene Ther 8, 354–361 (2001). https://doi.org/10.1038/sj.gt.3301409

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