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Clearance of persistent hepatitis C virus infection in humanized mice using a claudin-1-targeting monoclonal antibody

Nature Biotechnology volume 33, pages 549–554 (2015)Cite this article

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Abstract

Hepatitis C virus (HCV) infection is a leading cause of liver cirrhosis and cancer1. Cell entry of HCV2 and other pathogens3,4,5 is mediated by tight junction (TJ) proteins, but successful therapeutic targeting of TJ proteins has not been reported yet. Using a human liver–chimeric mouse model6, we show that a monoclonal antibody specific for the TJ protein claudin-1 (ref. 7) eliminates chronic HCV infection without detectable toxicity. This antibody inhibits HCV entry, cell-cell transmission and virus-induced signaling events. Antibody treatment reduces the number of HCV-infected hepatocytes in vivo, highlighting the need for de novo infection by means of host entry factors to maintain chronic infection. In summary, we demonstrate that an antibody targeting a virus receptor can cure chronic viral infection and uncover TJ proteins as targets for antiviral therapy.

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Figure 1: Human CLDN1 expression and tight junction ultrastructure in the livers of human chimeric mice.
Figure 2: Prevention and clearance of chronic HCV infection using a CLDN1-specific mAb in vivo.
Figure 3: CLDN1-specific mAb impairs HCV-induced host cell signaling.
Figure 4: CLDN1-specific mAb leads to elimination of HCV-infected cells from the livers of human chimeric mice in a dose- and time-dependent manner.

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Acknowledgements

This work was supported by the European Union (ERC-2008-AdG-233130-HEPCENT, ERC-2010-StG-260767-ncRNAVIR, INTERREG-IV-Rhin Supérieur-FEDER-Hepato-Regio-Net 2009 and 2012), ANRS (ANRS 2009/183, 2009/136, 2011/132, 2012/239, 2013/108), ANR (Laboratoires d'excellence ANR-10-LABX-0028_HEPSYS and ANR-10-LABX-36 netRNA), Fondation ARC pour la recherche (NanoISI and TheraHCC IHUARC IHU201301187), Institut Hospitalo-Universitaire (IHU) Strasbourg, the Wilhelm Sander Foundation, Région Alsace, Institut National du Cancer, the Institut National de la Santé et de la Recherche Médicale, Centre National de la Recherche Scientifique, Université de Strasbourg, the Ghent University (GOA 01G01712) and the Research Foundation—Flanders (projects 1500910N and G052112N). We are grateful to S. Ito (Harvard Medical School) for electron microscopy studies, F.-L. Cosset (Inserm U1111, ENS Lyon, France) and J. Ball (University of Nottingham, Nottingham, UK) for retroviral vectors for HCVpp production, F. Chisari (The Scripps Research Institute, La Jolla, CA, USA) for the gift of Huh7.5.1 cells, A. Patel (MRC Virology Unit, Glasgow, UK) for E2-specific mAb AP33 and Huh7.5-GFP cells, S. Foung (Stanford Blood Center, Palo Alto, CA, USA) for E2-specific mAb CHB-23 and C.M. Rice and M. Evans (Rockefeller University and Mount Sinai School of Medicine, New York) for providing human and mouse CLDN1 expression constructs as well as Huh7.5 cells. We acknowledge S. Durand, L. Heydmann, E. Soulier, J. Barths, N. Brignon, S. Pernot (Inserm U1110, Strasbourg), O. Wendling and N. Messadeq (Institut Clinique de la Souris - ICS, Illkirch), C. Valencia (PCBIS, Illkirch), S. Kallis (University of Heidelberg, Germany) for technical work, F. Grunert and J. Thompson (Aldevron, Freiburg) for helpful discussions, H. Jacob and M.F. Champy (ICS, Illkirch) for histopathological, hematological and biochemical analyzes, P. Bachellier (Strasbourg University Hospitals) for providing liver resections for isolation of primary human hepatocytes, the Laboratoire Schuh—groupement Bio67, Strasbourg and the Plateau Technique de Microbiologie, Laboratoire de Virologie (S. Fafi-Kremer and F. Stoll-Keller), University Hospital Strasbourg for performing viral load analyses, and the IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire) microarray and sequencing platform, member of the France Génomique program, for the sequencing of our libraries. Part of the animal experiments was carried out within the small animal exploration facility Cardiex (Nantes), which is supported by the GIS-IBiSA (Groupement d'Intérêt Scientifique – Infrastructure en Biologie Santé et Agronomie) program.

Author information

Author notes

  1. Fei Xiao and Joachim Lupberger: These authors contributed equally to this work.

  2. Eric Robinet and Thomas F Baumert: These authors jointly supervised this work.

Authors and Affiliations

  1. Institut National de la Santé et de la Recherche Médicale, Unité 1110, Institut de Recherche sur les Maladies Virales et Hépatiques, Strasbourg, France

    Laurent Mailly, Fei Xiao, Joachim Lupberger, Céline Leboeuf, Isabel Fofana, Christine Thumann, Simonetta Bandiera, Patrick Pessaux, Mirjam B Zeisel, Eric Robinet & Thomas F Baumert

  2. Université de Strasbourg, Strasbourg, France

    Laurent Mailly, Fei Xiao, Joachim Lupberger, Céline Leboeuf, Isabel Fofana, Christine Thumann, Simonetta Bandiera, Erika Girardi, Béatrice Chane-Woon-Ming, Patrick Pessaux, Pascal Villa, Sébastien Pfeffer, Mirjam B Zeisel, Eric Robinet & Thomas F Baumert

  3. Hepatitis C Research Group, Institute for Biomedical Research, University of Birmingham, Birmingham, UK

    Garrick K Wilson, Christopher Davis, Helen J Harris, Christopher J Mee, Nicola Fletcher & Jane A McKeating

  4. Institut National de la Santé et de la Recherche Médicale, Unité 913, Nantes, France

    Philippe Aubert & Michel Neunlist

  5. Université de Nantes, Nantes, France

    Philippe Aubert & Michel Neunlist

  6. Institut des Maladies de l'Appareil Digestif, CHU Nantes, Hôpital Hôtel-Dieu, Nantes, France

    Philippe Aubert & Michel Neunlist

  7. Department of Biomedicine, Hepatology Laboratory, University of Basel, Basel, Switzerland

    François H T Duong, Diego Calabrese & Markus H Heim

  8. Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

    Marc Lütgehetmann, Tassilo Volz & Maura Dandri

  9. Architecture et Réactivité de l'ARN, Institut de Biologie Moléculaire et Cellulaire du CNRS, Strasbourg, France

    Erika Girardi, Béatrice Chane-Woon-Ming & Sébastien Pfeffer

  10. Electron Microscopy Facility, Harvard Medical School, Boston, Massachusetts, USA

    Maria Ericsson

  11. Department of Infectious Diseases, Molecular Virology, Heidelberg University, Heidelberg, Germany

    Ralf Bartenschlager

  12. German Centre for Infection Research, Heidelberg University, Heidelberg, Germany

    Ralf Bartenschlager

  13. Pôle Hépato-Digestif, Institut Hospitalo-Universitaire, Hôpitaux Universitaires de Strasbourg, Strasbourg, France

    Patrick Pessaux & Thomas F Baumert

  14. Center for Vaccinology, Ghent University, Ghent, Belgium

    Koen Vercauteren & Philip Meuleman

  15. Plateforme de Chimie Biologique Intégrative de Strasbourg, Medalis, UMS 3286 CNRS-UdS & FMTS, Illkirch, France

    Pascal Villa

  16. Institute for Medical Informatics and Biometry, Medical Faculty, Technische Universität Dresden, Dresden, Germany

    Lars Kaderali

Authors
  1. Laurent Mailly
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  2. Fei Xiao
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Contributions

T.F.B. initiated and supervised the study. T.F.B., E.R., J.A.M., M.N., M.B.Z., M.H.H., R.B., S.P., P.M., P.V. and J.L. designed experiments and analyzed data. L.M., P.A., K.V. and E.R. performed in vivo experiments and analyzed data. L.M., F.X., J.L., S.B., G.K.W., P.A., F.H.T.D., D.C., C.L., M.E., I.F., C.D., H.J.H., C.J.M., C.T., E.G., B.C.-W.-M., N.F., M.B.Z. and L.K. performed ex vivo and in vitro experiments and analyzed data. R.B., P.P. and P.M. provided key reagents. M.D., M.L. and T.V. produced chimeric uPA-SCID mice. L.M., J.L., S.B., M.B.Z., J.A.M., E.R. and T.F.B. wrote the manuscript.

Corresponding author

Correspondence to Thomas F Baumert.

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Competing interests

Inserm, the University of Strasbourg and Genovac/Aldevron Freiburg have filed a patent application on monoclonal anti-claudin 1 antibodies for the inhibition of hepatitis C virus infection (US Patent # 8,518,408; WO2010034812; PCT/EP2009/062449). T.F.B. has served as a scientific advisor for Gilead, Biotest and Vironexx.

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Mailly, L., Xiao, F., Lupberger, J. et al. Clearance of persistent hepatitis C virus infection in humanized mice using a claudin-1-targeting monoclonal antibody. Nat Biotechnol 33, 549–554 (2015). https://doi.org/10.1038/nbt.3179

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