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Coagulation factor X shields adenovirus type 5 from attack by natural antibodies and complement

Nature Medicine volume 19, pages 452–457 (2013)Cite this article

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Abstract

Adenovirus type 5 (Ad5) specifically binds coagulation factor X (FX), and FX is normally essential for intravenously injected Ad5 vectors to transduce the liver. We demonstrate that the ability of FX to enhance liver transduction by Ad5 vectors is due to an unexpected ability of FX to protect Ad5 from attack by the classical complement pathway. In vitro, naive mouse serum neutralized Ad5 when FX was blocked from binding Ad5. This neutralization was mediated by natural IgM and the classical complement pathway. In vivo, FX was essential for Ad5 vectors to transduce the livers of wild-type mice, but FX was not required for liver transduction in mice that lack antibodies, C1q or C4. We conclude that Ad5 recruits FX as a defense against complement and that the sensitivity of Ad5 to inactivation by complement must be taken into account when designing vectors for systemic gene therapy.

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Figure 1: Vitamin K–dependent coagulation factors are required for liver transduction in WT C57BL/6 mice but not in Rag1−/− (B and T cell–deficient) mice.
Figure 2: B cells and natural IgM determine whether coagulation factors are required for liver transduction.
Figure 3: The classical complement pathway restricts liver transduction when coagulation factors are absent.
Figure 4: FX in mouse serum shields Ad5 from neutralization by complement.
Figure 5: FX blocks the ability of Ad5 vectors to activate C3.
Figure 6: Natural antibodies and complement inhibit transduction of nonhepatic organs when Ad5 vectors are unprotected by coagulation factors.

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Acknowledgements

Funding was provided by the US Food and Drug Administration (FDA), including the FDA's Critical Path program. This project was supported in part by fellowships administered by the Oak Ridge Institute for Science and Education. We thank M. Barry (Mayo Clinic) for providing vectors, H. Mizuguchi (Osaka University) for providing plasmid pAdHM4-CMVL1 and M. Diamond (Washington University) for providing C1qa−/− mice. We thank M. Shlomchik (Yale University), S. Epstein (FDA) and J. Misplon (FDA) for providing JHD and mIg Tg mice. We thank the Center for Biologics Evaluation and Research animal facility staff for outstanding support. We thank S. Epstein, C. Kimchi-Sarfaty, G. Price and C. Wiethoff for helpful discussions or comments on the manuscript.

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Authors and Affiliations

  1. Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA

    Zhili Xu, Qi Qiu, Jie Tian, Jeffrey S Smith, Gina M Conenello & Andrew P Byrnes

  2. Department of Biochemistry, Meiji Pharmaceutical University, Tokyo, Japan

    Takashi Morita

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Contributions

Z.X. discovered that coagulation factors are not required for liver transduction. A.P.B., Z.X., Q.Q., J.T. and J.S.S. designed experiments. Z.X. performed all in vivo experiments, collected serum and plasma, purified IgM, constructed AdHVR7, grew vectors, and performed quantitative PCR, western blotting and Octet RED assays. Q.Q. performed quantitative PCR and conducted in vitro transduction and neutralization studies. J.T. conducted complement activity assays and grew vectors. J.S.S. conducted cell-binding assays. G.M.C. conducted transduction assays. T.M. purified X-bp. A.P.B. bred mice. A.P.B. wrote the majority of the manuscript, and all authors participated in the preparation of the manuscript.

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Correspondence to Andrew P Byrnes.

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Xu, Z., Qiu, Q., Tian, J. et al. Coagulation factor X shields adenovirus type 5 from attack by natural antibodies and complement. Nat Med 19, 452–457 (2013). https://doi.org/10.1038/nm.3107

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