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Detection of Neutralizing Antibodies with HCV Pseudoparticles (HCVpp)

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Hepatitis C

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 510))

Abstract

Infectious HCV pseudoparticles (HCVpp) can be assembled by display of unmodified and functional HCV glycoproteins on retroviral and lentiviral core particles. HCVpp have been shown to mimic the early infection steps of parental HCV. The presence of a marker gene packaged within these HCV pseudoparticles allows reliable and fast determination of infectivity mediated by the HCV glycoproteins. With this highly flexible system, E1E2 from a broad range of HCV strains can be investigated, including autologous HCV strains from patients’ virus, and it has allowed careful investigation of the humoral response to HCV.

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References

  1. Shimizu, Y. K., Igarashi, H., Kiyohara, T., Cabezon, T., Farci, P., Purcell, R. H., et al. (1996) A hyperimmune serum against a synthetic peptide corresponding to the hypervariable region 1 of hepatitis C virus can prevent viral infection in cell cultures. Virology 223, 409–412.

    Article  CAS  PubMed  Google Scholar 

  2. Shimizu, Y. K., Hijikata, M., Iwamoto, A., Alter, H. J., Purcell, R. H., and Yoshikura, H. (1994) Neutralizing antibodies against hepatitis-C virus and the emergence of neutralization escape mutant viruses. J. Virol. 68, 1494–1500.

    CAS  PubMed  Google Scholar 

  3. Farci, P., Alter, H. J., Wong, D. C., Miller, R. H., Govindarajan, S., Engle, R., et al. (1994) Prevention of hepatitis-C virus infection in chimpanzees after antibody-mediated in vitro neutralization. Proc. Natl. Acad. Sci. USA 91, 7792–7796.

    Article  CAS  PubMed  Google Scholar 

  4. Farci, P., Shimoda, A., Wong, D., Cabezon, T., Gioannis, D. D., Strazzera, A., et al. (1996) Prevention of hepatitis C virus infection in chimpanzees by hyperimmune serum against the hypervariable region 1 of the envelope 2 protein. Proc. Natl. Acad. Sci. USA 93, 15394–15399.

    Article  CAS  PubMed  Google Scholar 

  5. Bartosch, B., Dubuisson, J., and Cosset, F. L. (2003) Infectious hepatitis C virus pseudo-particles containing functional E1-E2 envelope protein complexes. J. Exp. Med. 197, 633–642.

    Article  CAS  PubMed  Google Scholar 

  6. Hsu, M., Zhang, J., Flint, M., Logvinoff, C., Cheng-Mayer, C., Rice, C. M., et al. (2003) Hepatitis C virus glycoproteins mediate pH-dependent cell entry of pseudotyped retroviral particles. Proc. Natl. Acad. Sci. USA 100, 7271–7276.

    Article  CAS  PubMed  Google Scholar 

  7. Bartosch, B., Bukh, J., Meunier, J. C., Granier, C., Engle, R. E., Blackwelder, W. C., et al. (2003) In vitro assay for neutralizing antibody to hepatitis C virus: Evidence for broadly conserved neutralization epitopes. Proc. Natl. Acad. Sci. USA 100, 14199–14204.

    Article  CAS  PubMed  Google Scholar 

  8. Logvinoff, C., Major, M. E., Oldach, D., {Heyward,} S., Talal, A., Balfe, P., et al. (2004) Neutralizing antibody response during acute and chronic hepatitis C virus infection. Proc. Natl. Acad. Sci. USA 101, 10149–10154.

    Article  CAS  PubMed  Google Scholar 

  9. Lavillette, D., Morice, Y., Germanidis, G., Donot, P., Soulier, A., Pagkalos, E., et al. (2005) Human serum facilitates hepatitis C virus infection, and neutralizing responses inversely correlate with viral replication kinetics at the acute phase of hepatitis C virus infection. J. Virol. 79, 6023–6034.

    Article  CAS  PubMed  Google Scholar 

  10. Pestka, J. M., Zeisel, M. B., Blaser, E., {Schurmann,} P., Bartosch, B., Cosset, F. L., et al. (2007) Rapid induction of virus-neutralizing antibodies and viral clearance in a single-source outbreak of hepatitis C. Proc. Natl. Acad. Sci. USA 104, 6025–6030.

    Article  CAS  PubMed  Google Scholar 

  11. Fukumoto, T., Berg, T., Ku, Y., Bechstein, W. O., Knoop, M., Lemmens, H. P., et al. (1996) Viral dynamics of hepatitis C early after orthotopic liver transplantation: evidence for rapid turnover of serum virions. Hepatology 24, 1351–1354.

    Article  CAS  PubMed  Google Scholar 

  12. Neumann, A. U., Lam, N. P., Dahari, H., Gretch, D. R., Wiley, T. E., Layden, T. J., et al. (1998) Hepatitis C viral dynamics in vivo and the antiviral efficacy of interferon-alpha therapy. Science 282, 103–107.

    Article  CAS  PubMed  Google Scholar 

  13. Gomez, J., Martell, M., Quer, J., Cabot, B., and Esteban, J. I. (1999) Hepatitis C viral quasispecies. J. Viral. Hepat. 6, 3–16.

    Article  CAS  PubMed  Google Scholar 

  14. Pawlotsky, J. M. (2003) Hepatitis C virus genetic variability: pathogenic and clinical implications. Clin. Liver Dis. 7, 45–66.

    Article  PubMed  Google Scholar 

  15. Lavillette, D., Tarr, A. W., Voisset, C., Donot, P., Bartosch, B., Bain, C., et al. (2005) Characterization of host-range and cell entry properties of the major genotypes and subtypes of hepatitis C virus. Hepatology 41, 265–274.

    Article  PubMed  Google Scholar 

  16. Meunier, J. C., Engle, R. E., Faulk, K., Zhao, M., Bartosch, B., Alter, H., et al. (2005) Evidence for cross-genotype neutralization of hepatitis C virus pseudo-particles and enhancement of infectivity by apolipoprotein C1. Proc. Natl. Acad. Sci. USA 102, 4560–4565.

    Article  CAS  PubMed  Google Scholar 

  17. Owsianka, A., Tarr, A. W., Juttla, V. S., Lavillette, D., Bartosch, B., Cosset, F. L., et al. (2005) Monoclonal antibody AP33 defines a broadly neutralizing epitope on the hepatitis C virus E2 envelope glycoprotein. J. Virol. 79, 11095–11104.

    Article  CAS  PubMed  Google Scholar 

  18. McKeating, J. A., Zhang, L. Q., Logvinoff, C., Flint, M., Zhang, J., Yu, J., et al. (2004) Diverse hepatitis C virus glycoproteins mediate viral infection in a CD81-dependent manner. J. Virol. 78, 8496–8505.

    Article  CAS  PubMed  Google Scholar 

  19. Keck, Z. Y., Xia, J., Cai, Z., Li, T. K., Owsianka, A. M., Patel, A. H., et al. (2007) Immunogenic and functional organization of hepatitis C virus (HCV) glycoprotein E2 on infectious HCV virions. J. Virol. 81, 1043–1047.

    Article  CAS  PubMed  Google Scholar 

  20. Steinmann, D., Barth, H., Gissler, B., Schurmann, P., Adah, M. I., Gerlach, J. T., et al. (2004) Inhibition of hepatitis C virus-like particle binding to target cells by antiviral antibodies in acute and chronic hepatitis C. J. Virol. 78, 9030–9040.

    Google Scholar 

  21. Bartosch, B., Bukh, J., Meunier, J. C., Granier, C., Engle, R. E., Blackwelder, W. C., et al. (2003) In vitro assay for neutralizing antibody to hepatitis C virus: evidence for broadly conserved neutralization epitopes. Proc. Natl. Acad. Sci. USA 100, 14199–14204.

    Article  CAS  PubMed  Google Scholar 

  22. Netski, D. M., Mosbruger, T., Depla, E., Maertens, G., Ray, S. C., Hamilton, R. G., et al. (2005) Humoral immune response in acute hepatitis C virus infection. Clin. Infect. Dis. 41, 667–675.

    Article  CAS  PubMed  Google Scholar 

  23. von Hahn, T., Yoon, J. C., Alter, H., Rice, C. M., Rehermann, B., Balfe, P., et al. (2007) Hepatitis C virus continuously escapes from neutralizing antibody and T-cell responses during chronic infection in vivo. Gastroenterology 132, 667–678.

    Article  Google Scholar 

  24. Bartosch, B., Verney, G., Dreux, M., Donot, P., Morice, Y., Penin, F., et al. (2005) An interplay between hypervariable region 1 of the hepatitis C virus E2 glycoprotein, the scavenger receptor BI, and high-density lipoprotein promotes both enhancement of infection and protection against neutralizing antibodies. J. Virol. 79, 8217–8229.

    Article  CAS  PubMed  Google Scholar 

  25. Dreux, M., Pietschmann, T., Granier, C., Voisset, C., Ricard-Blum, S., Mangeot, P. E., et al. (2006) High density lipoprotein inhibits hepatitis C virus-neutralizing antibodies by stimulating cell entry via activation of the scavenger receptor BI. J. Biol. Chem. 281, 18285–18295.

    Article  CAS  PubMed  Google Scholar 

  26. Voisset, C., Op de Beeck, A., Horellou, P., Dreux, M., Gustot, T., Duverlie, G., et al. (2006) High-density lipoproteins reduce the neutralizing effect of hepatitis C virus (HCV)-infected patient antibodies by promoting HCV entry. J. Gen. Virol. 87, 2577–2581.

    Article  CAS  PubMed  Google Scholar 

  27. Bartosch, B., Vitelli, A., Granier, C., {Goujon,} C., Dubuisson, J., Pascale, S., et al. (2003) Cell entry of hepatitis C virus requires a set of co-receptors that include the CD81 tetraspanin and the SR-B1 scavenger receptor. J. Biol. Chem. 278, 41624–41630.

    Article  CAS  PubMed  Google Scholar 

  28. Maillard, P., Huby, T., Andreo, U., Moreau, M., Chapman, J., and Budkowska, A. (2006) The interaction of natural hepatitis C virus with human scavenger receptor SR-BI/Cla1 is mediated by ApoB-containing lipoproteins. FASEB J. 20, 735–737.

    CAS  PubMed  Google Scholar 

  29. Scarselli, E., Ansuini, H., Cerino, R., Roccasecca, R., Acali, S., Filocamo, G., et al. (2002) The human scavenger receptor class B type I is a novel candidate receptor for the hepatitis C virus. EMBO J. 21, 5017–5025.

    Article  CAS  PubMed  Google Scholar 

  30. Kapadia, S. B., Barth, H., Baumert, T., McKeating, J. A., and Chisari, F. V. (2006) Initiation of hepatitis C virus infection is dependent on cholesterol and cooperativity between CD81 and scavenger receptor B type I. J. Virol. 81, 374–383.

    Article  PubMed  Google Scholar 

  31. Krieger, M. (2001) Scavenger receptor class B type I is a multiligand HDL receptor that influences diverse physiologic systems. J. Clin. Invest. 108, 793–797.

    CAS  PubMed  Google Scholar 

  32. Sandrin, V., and Cosset, F. L. (2006) Intracellular versus cell surface assembly of retroviral pseudotypes is determined by the cellular localization of the viral glycoprotein, its capacity to interact with Gag, and the expression of the Nef protein. J. Biol. Chem. 281, 528–542.

    Article  CAS  PubMed  Google Scholar 

  33. Nakabayashi, H., Taketa, K., Miyano, K., Yamane, T., and Sato, J. (1982) Growth of human hepatoma-cell lines with differentiated functions in chemically defined medium. Cancer Res. 42, 3858–3863.

    CAS  PubMed  Google Scholar 

  34. Schofield, D. J., Bartosch, B., Shimizu, Y. K., Allander, T., Alter, A. J., Emerson, S. U., et al. (2005) Human monoclonal antibodies that react with the E2 glycoprotein of hepatitis C virus and possess neutralizing activity. Hepatology 42, 1055–1062.

    Article  CAS  PubMed  Google Scholar 

  35. Voisset, C., Callens, N., Blanchard, E., Op De Beeck, A., Dubuisson, J., and Vu-Dac, N. (2005) High density lipoproteins facilitate hepatitis C virus entry through the scavenger receptor class B type I. J. Biol. Chem. 280, 7793–7799.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We are grateful to our coworkers and colleagues for encouragement and advice. Our work is supported by INSERM, ENS Lyon, and CNRS and by grants from the Ligue Nationale Contre le Cancer, the European Union (LSHB-CT-2004-005246), and the Agence Nationale de Recherches sur le SIDA et les Hépatites Virales (ANRS).

Author information

Authors and Affiliations

  1. Universit de Lyon, (UCB-Lyon1), Lyon, IFR128, France

    Marlène Dreux

  2. ENS de Lyon, Lyon, France

    François-Loïc Cosset

Authors
  1. Marlène Dreux
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  2. François-Loïc Cosset
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Editor information

Editors and Affiliations

  1. Florida State University, Tallahassee, FL, USA

    Hengli Tang

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Dreux, M., Cosset, FL. (2009). Detection of Neutralizing Antibodies with HCV Pseudoparticles (HCVpp). In: Tang, H. (eds) Hepatitis C. Methods in Molecular Biology™, vol 510. Humana Press. https://doi.org/10.1007/978-1-59745-394-3_32

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  • DOI: https://doi.org/10.1007/978-1-59745-394-3_32

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-970-3

  • Online ISBN: 978-1-59745-394-3

  • eBook Packages: Springer Protocols

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