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A live, impaired-fidelity coronavirus vaccine protects in an aged, immunocompromised mouse model of lethal disease

Nature Medicine volume 18, pages 1820–1826 (2012)Cite this article

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Abstract

Live, attenuated RNA virus vaccines are efficacious but subject to reversion to virulence. Among RNA viruses, replication fidelity is recognized as a key determinant of virulence and escape from antiviral therapy; increased fidelity is attenuating for some viruses. Coronavirus (CoV) replication fidelity is approximately 20-fold greater than that of other RNA viruses and is mediated by a 3′→5′ exonuclease (ExoN) activity that probably functions in RNA proofreading. In this study we demonstrate that engineered inactivation of severe acute respiratory syndrome (SARS)-CoV ExoN activity results in a stable mutator phenotype with profoundly decreased fidelity in vivo and attenuation of pathogenesis in young, aged and immunocompromised mice. The ExoN inactivation genotype and mutator phenotype are stable and do not revert to virulence, even after serial passage or long-term persistent infection in vivo. ExoN inactivation has potential for broad applications in the stable attenuation of CoVs and, perhaps, other RNA viruses.

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Figure 1: The nsp14 ExoN mutator virus in a virulent mouse-adapted SARS-CoV isogenic background.
Figure 2: Weight loss and lung titer in BALB/c mice.
Figure 3: Weight loss and lung titer in young immunocompromised mice.
Figure 4: Mutation accumulation in infected SCID mice at 30 d.p.i.
Figure 5: Virulence of passaged MA-ExoN and MAwt viruses.
Figure 6: MA-ExoN vaccination protects from lethal challenge.

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Acknowledgements

The authors thank R. Halpin, C. Town (US National Institutes of Health Microbial Genome Sequencing Contract HHSN272200900007C) and X. Lu for their assistance in sequencing in vitro isolates. This work was funded by US National Institutes of Health grants U54-AI057157 (SERCEB; R.S.B. and M.R.D.), AI075297 (R.S.B.) and 5F32AI080148 (R.L.G.).

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Authors and Affiliations

  1. Department of Epidemiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA

    Rachel L Graham & Ralph S Baric

  2. Department of Pediatrics, Vanderbilt University, Nashville, Tennessee, USA

    Michelle M Becker, Lance D Eckerle & Mark R Denison

  3. Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA

    Meagan Bolles & Ralph S Baric

  4. Department of Pathology, Microbiology and Immunology, Vanderbilt University, Nashville, Tennessee, USA

    Mark R Denison

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  1. Rachel L Graham
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  3. Lance D Eckerle
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  4. Meagan Bolles
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Contributions

R.L.G. designed and performed experiments, analyzed data, and wrote and edited the paper. M.M.B., L.D.E. and M.B. performed experiments, analyzed data and read the paper. M.R.D. and R.S.B. designed experiments, analyzed data, and wrote and edited the paper.

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Correspondence to Ralph S Baric.

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Graham, R., Becker, M., Eckerle, L. et al. A live, impaired-fidelity coronavirus vaccine protects in an aged, immunocompromised mouse model of lethal disease. Nat Med 18, 1820–1826 (2012). https://doi.org/10.1038/nm.2972

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