An alpaca nanobody inhibits hepatitis C virus entry and cell-to-cell transmission

Authors

  • Alexander W. Tarr,

    1. School of Molecular Medical Sciences, The University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom
    2. The Nottingham Digestive Diseases Centre Biomedical Research Unit, The University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom
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  • Pierre Lafaye,

    1. Institut Pasteur, Plateforme d'Ingénierie des Anticorps, Proteopole-Departement de Biologie Structurale et Chimie, Paris, France
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  • Luke Meredith,

    1. Hepatitis C Research Group, Centre for Human Virology, University of Birmingham, Birmingham, United Kingdom
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  • Laurence Damier-Piolle,

    1. Institut Pasteur, Unité de Virologie Structurale, Departement Virologie, Paris, France
    2. CNRS URA 3569, Paris, France
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  • Richard A. Urbanowicz,

    1. School of Molecular Medical Sciences, The University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom
    2. The Nottingham Digestive Diseases Centre Biomedical Research Unit, The University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom
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  • Annalisa Meola,

    1. Institut Pasteur, Unité de Virologie Structurale, Departement Virologie, Paris, France
    2. CNRS URA 3569, Paris, France
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  • Jean-Luc Jestin,

    1. Institut Pasteur, Unité de Virologie Structurale, Departement Virologie, Paris, France
    2. CNRS URA 3569, Paris, France
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  • Richard J. P. Brown,

    1. School of Molecular Medical Sciences, The University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom
    2. The Nottingham Digestive Diseases Centre Biomedical Research Unit, The University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom
    Current affiliation:
    1. Twincore, Zentrum für Experimentelle und Klinische Infektionsforschung GmbH, Hannover, Germany
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  • Jane A. McKeating,

    1. Hepatitis C Research Group, Centre for Human Virology, University of Birmingham, Birmingham, United Kingdom
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  • Felix A. Rey,

    1. Institut Pasteur, Unité de Virologie Structurale, Departement Virologie, Paris, France
    2. CNRS URA 3569, Paris, France
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  • Jonathan K. Ball,

    Corresponding author
    1. The Nottingham Digestive Diseases Centre Biomedical Research Unit, The University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom
    • School of Molecular Medical Sciences, The University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom
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  • Thomas Krey

    Corresponding author
    1. Institut Pasteur, Unité de Virologie Structurale, Departement Virologie, Paris, France
    2. CNRS URA 3569, Paris, France
    • School of Molecular Medical Sciences, The University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom
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  • Potential conflict of interest: Nothing to report.

  • This work was funded by grants from the UK Medical Research Council, ANRS, and recurrent funding from Institut Pasteur, CNRS, and Merck-Serono (to F. A. R.).

Address reprint requests to: Thomas Krey, Institut Pasteur, Structural Virology Unit, 25-28 Rue du Docteur Roux, 75015 Paris, France. E-mail: tkrey@pasteur.fr; fax: (33)-145688993; or

Jonathan Ball, Virus Research Group, A Floor, West Block QMC, Queen's Medical Centre, Nottingham NG7 2UH, UK. E-mail: jonathan.ball@nottingham.ac.uk; fax: (44)-1158230759.

Abstract

Severe liver disease caused by chronic hepatitis C virus is the major indication for liver transplantation. Despite recent advances in antiviral therapy, drug toxicity and unwanted side effects render effective treatment in liver-transplanted patients a challenging task. Virus-specific therapeutic antibodies are generally safe and well-tolerated, but their potential in preventing and treating hepatitis C virus (HCV) infection has not yet been realized due to a variety of issues, not least high production costs and virus variability. Heavy-chain antibodies or nanobodies, produced by camelids, represent an exciting antiviral approach; they can target novel highly conserved epitopes that are inaccessible to normal antibodies, and they are also easy to manipulate and produce. We isolated four distinct nanobodies from a phage-display library generated from an alpaca immunized with HCV E2 glycoprotein. One of them, nanobody D03, recognized a novel epitope overlapping with the epitopes of several broadly neutralizing human monoclonal antibodies. Its crystal structure revealed a long complementarity determining region (CD3) folding over part of the framework that, in conventional antibodies, forms the interface between heavy and light chain. D03 neutralized a panel of retroviral particles pseudotyped with HCV glycoproteins from six genotypes and authentic cell culture–derived particles by interfering with the E2-CD81 interaction. In contrast to some of the most broadly neutralizing human anti-E2 monoclonal antibodies, D03 efficiently inhibited HCV cell-to-cell transmission. Conclusion: This is the first description of a potent and broadly neutralizing HCV-specific nanobody representing a significant advance that will lead to future development of novel entry inhibitors for the treatment and prevention of HCV infection and help our understanding of HCV cell-to-cell transmission. (Hepatology 2013;53:932–939)

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