Gastroenterology

Gastroenterology

Volume 129, Issue 1, July 2005, Pages 234-245
Gastroenterology

Basic-liver, pancreas, and biliary tract
Mapping of the Hepatitis B Virus Attachment Site by Use of Infection-Inhibiting preS1 Lipopeptides and Tupaia Hepatocytes

https://doi.org/10.1053/j.gastro.2005.03.090Get rights and content

Background & aims: Studies on the early steps in the life cycle of hepatitis B virus have been hampered by the lack of readily available target cells. In this study, we mapped a defined virus attachment site to primary hepatocytes that is essential for infection. Methods: We used purified virus particles from human carrier plasma as an inoculum and primary cultures of tupaia hepatocytes as susceptible target cells and studied the inhibitory effect of amino-terminally acylated preS1-derived lipopeptides on infection interference. Results: Infectivity of virus could be blocked efficiently in this system by amino-terminally acylated peptides containing amino acids 2–18 from the preS1 domain. The addition of amino acids 28–48 enhanced the inhibitory capacity, whereas amino acids 49–78 did not contribute to inhibition. Myristoylated preS1 peptides 2–48 bound strongly to tupaia hepatocytes but not to nonhepatic cells or rodent hepatocytes and thereby inhibited infection even at concentrations of 1 nmol/L completely. Particles consisting only of the small hepatitis B surface protein—the active component of current hepatitis B vaccines—did not bind at all to tupaia hepatocytes, but the addition of the preS1 domain to the particles allowed binding. Conclusions: The preS1 sequence 2–48 mediates attachment of the virus to its target cells, whereas the small surface protein seems to be involved in other steps. These findings indicate that the current subunit hepatitis B vaccines may be improved by the addition of distinct preS1 epitopes. Moreover, preS1 lipopeptides are promising candidates for specific antiviral therapy against hepatitis B infections.

Section snippets

Isolation and Purification of Hepatitis B Virus Virions and Subviral Particles From Plasma of Hepatitis B Virus–Infected Patients

HBV and natural HBs antigen (HBsAg; genotype D; HBsAg subtype ayw2) were isolated from hepatitis B early antigen (HBeAg)-positive plasma of 2 asymptomatic chronic HBV carriers with normal transaminase levels in serum. One carrier (ID304) had 1.6 × 109 HBV genomes per milliliter and 55 μg/mL HBsAg, and the second (ID259) had 2 × 109 HBV genomes per milliliter and 10 μg/mL HBsAg.

The purification was performed as previously described,19 with some modifications: HBV and subviral particles from 350

Inhibition of Hepatitis B Virus Infection of Primary Tupaia Hepatocytes by Myristoylated Peptides

For HBV infection of tupaia hepatocytes, we used highly purified plasma-derived HBV particles that contained less than 10% subviral HBsAg particles. We determined HBeAg secretion of infected cells as a marker of an established infection, because HBeAg is a nonstructural viral protein and therefore not present in the purified viral input. The amount of secreted HBeAg is approximately proportional to the amount of infectious HBV in the inoculum.10

We have shown previously that infection of primary

Discussion

Our previous study10 suggested that both the preS1 and S-domains of the HBs protein could be involved in the attachment and uptake of HBV by primary tupaia hepatocyte cultures because certain MAbs against both the preS1 and the S-domain inhibited the infection process very efficiently. The data of this study fully support the importance of the preS1 domain for the infection process at the level of attachment. Surprisingly, a role of the SHBs or the HBsAg epitopes in the S-domain for the

References (34)

  • A.R. Neurath et al.

    Identification and chemical synthesis of a host cell receptor binding site on hepatitis B virus

    Cell

    (1986)
  • P. Pontisso et al.

    Identification of an attachment site for human liver plasma membranes on hepatitis B virus particles

    Virology

    (1989)
  • M.D. Young et al.

    A multi-center controlled study of rapid hepatitis B vaccination using a novel triple antigen recombinant vaccine

    Vaccine

    (2001)
  • M.Y. Shapira et al.

    Rapid seroprotection against hepatitis B following the first dose of a Pre-S1/Pre-S2/S vaccine

    J Hepatol

    (2001)
  • S. Maurer-Stroh et al.

    Myristoylation of viral and bacterial proteins

    Trends Microbiol

    (2004)
  • M.H.V. van Regenmortel et al.

    Virus taxonomythe classification and nomenclature of viruses. The seventh report of the International Committee on Taxonomy of Viruses

    (2000)
  • D. Shouval

    Hepatitis B vaccines

    J Hepatol

    (2003)
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    Supported by Grants SFB 535/A2 (to W.H.G.) and UR72/4-1/4-2 (to S.U.) from the Deutsche Forschungsgemeinschaft and European Union Grant QLK2-2000-01476 (to K.S. and W.H.G.).

    1

    D.G. and S.U. contributed equally to this work.

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