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


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.


  1. Thomas, D.L. Global control of hepatitis C: where challenge meets opportunity. Nat. Med. 19, 850–858 (2013).

    Article  CAS  Google Scholar 

  2. Evans, M.J. et al. Claudin-1 is a hepatitis C virus co-receptor required for a late step in entry. Nature 446, 801–805 (2007).

    Article  CAS  Google Scholar 

  3. Cohen, C.J. et al. The coxsackievirus and adenovirus receptor is a transmembrane component of the tight junction. Proc. Natl. Acad. Sci. USA 98, 15191–15196 (2001).

    Article  CAS  Google Scholar 

  4. Fukumatsu, M. et al. Shigella targets epithelial tricellular junctions and uses a noncanonical clathrin-dependent endocytic pathway to spread between cells. Cell Host Microbe 11, 325–336 (2012).

    Article  CAS  Google Scholar 

  5. Che, P., Tang, H. & Li, Q. The interaction between claudin-1 and dengue viral prM/M protein for its entry. Virology 446, 303–313 (2013).

    Article  CAS  Google Scholar 

  6. Mercer, D.F. et al. Hepatitis C virus replication in mice with chimeric human livers. Nat. Med. 7, 927–933 (2001).

    Article  CAS  Google Scholar 

  7. Fofana, I. et al. Monoclonal anti-claudin 1 antibodies prevent hepatitis C virus infection of primary human hepatocytes. Gastroenterology 139, 953–964 (2010).

    Article  CAS  Google Scholar 

  8. Law, M. et al. Broadly neutralizing antibodies protect against hepatitis C virus quasispecies challenge. Nat. Med. 14, 25–27 (2008).

    Article  CAS  Google Scholar 

  9. Lupberger, J. et al. EGFR and EphA2 are host factors for hepatitis C virus entry and possible targets for antiviral therapy. Nat. Med. 17, 589–595 (2011).

    Article  CAS  Google Scholar 

  10. Meuleman, P. et al. A human monoclonal antibody targeting scavenger receptor class B type I precludes hepatitis C virus infection and viral spread in vitro and in vivo. Hepatology 55, 364–372 (2012).

    Article  CAS  Google Scholar 

  11. Shi, N. et al. Combination therapies with NS5A, NS3 and NS5B inhibitors on different genotypes of hepatitis C virus in human hepatocyte chimeric mice. Gut 62, 1055–1061 (2013).

    Article  CAS  Google Scholar 

  12. Harris, H.J., Wilson, G.K., Hubscher, S.G. & McKeating, J.A. Heterogeneous claudin-1 expression in human liver. Hepatology 57, 854–855 (2013).

    Article  CAS  Google Scholar 

  13. Fletcher, N.F. et al. Activated macrophages promote hepatitis C virus entry in a tumor necrosis factor-dependent manner. Hepatology 59, 1320–1330 (2014).

    Article  CAS  Google Scholar 

  14. Bukh, J. et al. Challenge pools of hepatitis C virus genotypes 1–6 prototype strains: replication fitness and pathogenicity in chimpanzees and human liver-chimeric mouse models. J. Infect. Dis. 201, 1381–1389 (2010).

    Article  CAS  Google Scholar 

  15. Pietschmann, T. et al. Construction and characterization of infectious intragenotypic and intergenotypic hepatitis C virus chimeras. Proc. Natl. Acad. Sci. USA 103, 7408–7413 (2006).

    Article  CAS  Google Scholar 

  16. Wakita, T. et al. Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nat. Med. 11, 791–796 (2005).

    Article  CAS  Google Scholar 

  17. Fofana, I. et al. Mutations that alter use of hepatitis C virus cell entry factors mediate escape from neutralizing antibodies. Gastroenterology 143, 223–233 (2012).

    Article  CAS  Google Scholar 

  18. Krieger, S.E. et al. Inhibition of hepatitis C virus infection by anti-claudin-1 antibodies is mediated by neutralization of E2–CD81-claudin-1 associations. Hepatology 51, 1144–1157 (2010).

    Article  CAS  Google Scholar 

  19. Dübel, S. & Reichert, J.M. (eds.). Handbook of Therapeutic Antibodies (Wiley-Blackwell, 2014).

  20. Harris, H.J. et al. Claudin association with CD81 defines hepatitis C virus entry. J. Biol. Chem. 285, 21092–21102 (2010).

    Article  CAS  Google Scholar 

  21. Suh, Y. et al. Claudin-1 induces epithelial-mesenchymal transition through activation of the c-Abl-ERK signaling pathway in human liver cells. Oncogene 32, 4873–4882 (2013).

    Article  CAS  Google Scholar 

  22. Brazzoli, M. et al. CD81 is a central regulator of cellular events required for hepatitis C virus infection of human hepatocytes. J. Virol. 82, 8316–8329 (2008).

    Article  CAS  Google Scholar 

  23. Diao, J. et al. Hepatitis C virus induces epidermal growth factor receptor activation via CD81 binding for viral internalization and entry. J. Virol. 86, 10935–10949 (2012).

    Article  CAS  Google Scholar 

  24. Zona, L. et al. HRas signal transduction promotes hepatitis C virus cell entry by triggering assembly of the host tetraspanin receptor complex. Cell Host Microbe 13, 302–313 (2013).

    Article  CAS  Google Scholar 

  25. Menzel, N. et al. MAP-kinase regulated cytosolic phospholipase A2 activity is essential for production of infectious hepatitis C virus particles. PLoS Pathog. 8, e1002829 (2012).

    Article  CAS  Google Scholar 

  26. Wieland, S. et al. Simultaneous detection of hepatitis C virus and interferon stimulated gene expression in infected human liver. Hepatology 59, 2121–2130 (2014).

    Article  CAS  Google Scholar 

  27. Neumann, A.U. et al. Hepatitis C viral dynamics in vivo and the antiviral efficacy of interferon-alpha therapy. Science 282, 103–107 (1998).

    Article  CAS  Google Scholar 

  28. Lan, L. et al. Hepatitis C virus infection sensitizes human hepatocytes to TRAIL-induced apoptosis in a caspase 9-dependent manner. J. Immunol. 181, 4926–4935 (2008).

    Article  CAS  Google Scholar 

  29. Haid, S. et al. Isolate-dependent use of Claudins for cell entry by hepatitis C virus. Hepatology 59, 24–34 (2014).

    Article  CAS  Google Scholar 

  30. Fofana, I. et al. Functional analysis of claudin-6 and claudin-9 as entry factors for hepatitis C virus infection of human hepatocytes by using monoclonal antibodies. J. Virol. 87, 10405–10410 (2013).

    Article  CAS  Google Scholar 

  31. Reynolds, G.M. et al. Hepatitis C virus receptor expression in normal and diseased liver tissue. Hepatology 47, 418–427 (2008).

    Article  Google Scholar 

  32. Koutsoudakis, G., Herrmann, E., Kallis, S., Bartenschlager, R. & Pietschmann, T. The level of CD81 cell surface expression is a key determinant for productive entry of hepatitis C virus into host cells. J. Virol. 81, 588–598 (2007).

    Article  CAS  Google Scholar 

  33. Harris, H.J. et al. CD81 and claudin 1 coreceptor association: role in hepatitis C virus entry. J. Virol. 82, 5007–5020 (2008).

    Article  CAS  Google Scholar 

  34. Piche, T. et al. Impaired intestinal barrier integrity in the colon of patients with irritable bowel syndrome: involvement of soluble mediators. Gut 58, 196–201 (2009).

    Article  CAS  Google Scholar 

  35. Zahid, M.N. et al. The postbinding activity of scavenger receptor class B type I mediates initiation of hepatitis C virus infection and viral dissemination. Hepatology 57, 492–504 (2013).

    Article  CAS  Google Scholar 

  36. Petersen, J. et al. Prevention of hepatitis B virus infection in vivo by entry inhibitors derived from the large envelope protein. Nat. Biotechnol. 26, 335–341 (2008).

    Article  CAS  Google Scholar 

  37. Dill, M.T. et al. Interferon-gamma-stimulated genes, but not USP18, are expressed in livers of patients with acute hepatitis C. Gastroenterology 143, 777–786 (2012).

    Article  CAS  Google Scholar 

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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.

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Authors and Affiliations



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.

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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).

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