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. 1992 Feb;66(2):1241–1245. doi: 10.1128/jvi.66.2.1241-1245.1992

Production of infectious hepatitis delta virus in vitro and neutralization with antibodies directed against hepatitis B virus pre-S antigens.

C Sureau 1, A M Moriarty 1, G B Thornton 1, R E Lanford 1
PMCID: PMC240836  PMID: 1309901

Abstract

Hepatitis delta virus (HDV) particles were produced in Huh7 human hepatoma cells by transfection with cloned hepatitis B virus (HBV) DNA and HDV cDNA. The particles were characterized by their buoyant density, the presence of encapsidated viral RNA, and their ability to infect primary cultures of chimpanzee hepatocytes. Successful infection was evidenced by the appearance of increasing amounts of intracellular HDV RNA after exposure to particles. Infection was prevented when particles were incubated with antibodies directed against synthetic peptides specific for epitopes of the pre-S1 or pre-S2 domains of the HBV envelope proteins before exposure to hepatocytes. These data demonstrate that HDV particles produced in vitro are infectious and indicate (i) that infectious particles are coated with HBV envelope proteins that contain the pre-S1 and pre-S2 regions, (ii) that epitopes of the pre-S1 and pre-S2 domains of HBV envelope proteins are exposed at the surface of HDV particles, and (iii) that antibodies directed against those epitopes have neutralizing activity against HDV.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bergmann K. F., Gerin J. L. Antigens of hepatitis delta virus in the liver and serum of humans and animals. J Infect Dis. 1986 Oct;154(4):702–706. doi: 10.1093/infdis/154.4.702. [DOI] [PubMed] [Google Scholar]
  2. Bonino F., Heermann K. H., Rizzetto M., Gerlich W. H. Hepatitis delta virus: protein composition of delta antigen and its hepatitis B virus-derived envelope. J Virol. 1986 Jun;58(3):945–950. doi: 10.1128/jvi.58.3.945-950.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonino F., Hoyer B., Shih J. W., Rizzetto M., Purcell R. H., Gerin J. L. Delta hepatitis agent: structural and antigenic properties of the delta-associated particle. Infect Immun. 1984 Mar;43(3):1000–1005. doi: 10.1128/iai.43.3.1000-1005.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Branch A. D., Robertson H. D. A replication cycle for viroids and other small infectious RNA's. Science. 1984 Feb 3;223(4635):450–455. doi: 10.1126/science.6197756. [DOI] [PubMed] [Google Scholar]
  5. Brown S. E., Howard C. R., Zuckerman A. J., Steward M. W. Affinity of antibody responses in man to hepatitis B vaccine determined with synthetic peptides. Lancet. 1984 Jul 28;2(8396):184–187. doi: 10.1016/s0140-6736(84)90479-3. [DOI] [PubMed] [Google Scholar]
  6. Bruss V., Ganem D. Mutational analysis of hepatitis B surface antigen particle assembly and secretion. J Virol. 1991 Jul;65(7):3813–3820. doi: 10.1128/jvi.65.7.3813-3820.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bruss V., Ganem D. The role of envelope proteins in hepatitis B virus assembly. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):1059–1063. doi: 10.1073/pnas.88.3.1059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chen P. J., Chen D. S., Chen C. R., Chen Y. Y., Chen H. M., Lai M. Y., Sung J. L. Delta infection in asymptomatic carriers of hepatitis B surface antigen: low prevalence of delta activity and effective suppression of hepatitis B virus replication. Hepatology. 1988 Sep-Oct;8(5):1121–1124. doi: 10.1002/hep.1840080525. [DOI] [PubMed] [Google Scholar]
  10. Chen P. J., Kalpana G., Goldberg J., Mason W., Werner B., Gerin J., Taylor J. Structure and replication of the genome of the hepatitis delta virus. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8774–8778. doi: 10.1073/pnas.83.22.8774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Diener T. O. Viroid processing: a model involving the central conserved region and hairpin I. Proc Natl Acad Sci U S A. 1986 Jan;83(1):58–62. doi: 10.1073/pnas.83.1.58. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eble B. E., Lingappa V. R., Ganem D. Hepatitis B surface antigen: an unusual secreted protein initially synthesized as a transmembrane polypeptide. Mol Cell Biol. 1986 May;6(5):1454–1463. doi: 10.1128/mcb.6.5.1454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eble B. E., MacRae D. R., Lingappa V. R., Ganem D. Multiple topogenic sequences determine the transmembrane orientation of the hepatitis B surface antigen. Mol Cell Biol. 1987 Oct;7(10):3591–3601. doi: 10.1128/mcb.7.10.3591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Emini E. A., Larson V., Eichberg J., Conard P., Garsky V. M., Lee D. R., Ellis R. W., Miller W. J., Anderson C. A., Gerety R. J. Protective effect of a synthetic peptide comprising the complete preS2 region of the hepatitis B virus surface protein. J Med Virol. 1989 May;28(1):7–12. doi: 10.1002/jmv.1890280103. [DOI] [PubMed] [Google Scholar]
  15. Ganem D., Varmus H. E. The molecular biology of the hepatitis B viruses. Annu Rev Biochem. 1987;56:651–693. doi: 10.1146/annurev.bi.56.070187.003251. [DOI] [PubMed] [Google Scholar]
  16. Gripon P., Diot C., Thézé N., Fourel I., Loreal O., Brechot C., Guguen-Guillouzo C. Hepatitis B virus infection of adult human hepatocytes cultured in the presence of dimethyl sulfoxide. J Virol. 1988 Nov;62(11):4136–4143. doi: 10.1128/jvi.62.11.4136-4143.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Itoh Y., Takai E., Ohnuma H., Kitajima K., Tsuda F., Machida A., Mishiro S., Nakamura T., Miyakawa Y., Mayumi M. A synthetic peptide vaccine involving the product of the pre-S(2) region of hepatitis B virus DNA: protective efficacy in chimpanzees. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9174–9178. doi: 10.1073/pnas.83.23.9174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jacob J. R., Eichberg J. W., Lanford R. E. In vitro replication and expression of hepatitis B virus from chronically infected primary chimpanzee hepatocytes. Hepatology. 1989 Dec;10(6):921–927. doi: 10.1002/hep.1840100605. [DOI] [PubMed] [Google Scholar]
  19. Krogsgaard K., Kryger P., Aldershvile J., Andersson P., Sørensen T. I., Nielsen J. O. Delta-infection and suppression of hepatitis B virus replication in chronic HBsAg carriers. Hepatology. 1987 Jan-Feb;7(1):42–45. doi: 10.1002/hep.1840070110. [DOI] [PubMed] [Google Scholar]
  20. Kuo M. Y., Chao M., Taylor J. Initiation of replication of the human hepatitis delta virus genome from cloned DNA: role of delta antigen. J Virol. 1989 May;63(5):1945–1950. doi: 10.1128/jvi.63.5.1945-1950.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kuo M. Y., Goldberg J., Coates L., Mason W., Gerin J., Taylor J. Molecular cloning of hepatitis delta virus RNA from an infected woodchuck liver: sequence, structure, and applications. J Virol. 1988 Jun;62(6):1855–1861. doi: 10.1128/jvi.62.6.1855-1861.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kuroki K., Floreani M., Mimms L. T., Ganem D. Epitope mapping of the PreS1 domain of the hepatitis B virus large surface protein. Virology. 1990 Jun;176(2):620–624. doi: 10.1016/0042-6822(90)90032-m. [DOI] [PubMed] [Google Scholar]
  23. Kuroki K., Russnak R., Ganem D. Novel N-terminal amino acid sequence required for retention of a hepatitis B virus glycoprotein in the endoplasmic reticulum. Mol Cell Biol. 1989 Oct;9(10):4459–4466. doi: 10.1128/mcb.9.10.4459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lanford R. E., Carey K. D., Estlack L. E., Smith G. C., Hay R. V. Analysis of plasma protein and lipoprotein synthesis in long-term primary cultures of baboon hepatocytes maintained in serum-free medium. In Vitro Cell Dev Biol. 1989 Feb;25(2):174–182. doi: 10.1007/BF02626175. [DOI] [PubMed] [Google Scholar]
  25. Negro F., Korba B. E., Forzani B., Baroudy B. M., Brown T. L., Gerin J. L., Ponzetto A. Hepatitis delta virus (HDV) and woodchuck hepatitis virus (WHV) nucleic acids in tissues of HDV-infected chronic WHV carrier woodchucks. J Virol. 1989 Apr;63(4):1612–1618. doi: 10.1128/jvi.63.4.1612-1618.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Neurath A. R., Kent S. B., Parker K., Prince A. M., Strick N., Brotman B., Sproul P. Antibodies to a synthetic peptide from the preS 120-145 region of the hepatitis B virus envelope are virus neutralizing. Vaccine. 1986 Mar;4(1):35–37. doi: 10.1016/s0264-410x(86)80001-9. [DOI] [PubMed] [Google Scholar]
  27. Neurath A. R., Kent S. B., Strick N. Location and chemical synthesis of a pre-S gene coded immunodominant epitope of hepatitis B virus. Science. 1984 Apr 27;224(4647):392–395. doi: 10.1126/science.6200931. [DOI] [PubMed] [Google Scholar]
  28. Neurath A. R., Seto B., Strick N. Antibodies to synthetic peptides from the preS1 region of the hepatitis B virus (HBV) envelope (env) protein are virus-neutralizing and protective. Vaccine. 1989 Jun;7(3):234–236. doi: 10.1016/0264-410x(89)90235-1. [DOI] [PubMed] [Google Scholar]
  29. Petcu D. J., Aldrich C. E., Coates L., Taylor J. M., Mason W. S. Suramin inhibits in vitro infection by duck hepatitis B virus, Rous sarcoma virus, and hepatitis delta virus. Virology. 1988 Dec;167(2):385–392. [PubMed] [Google Scholar]
  30. Petit M. A., Dubanchet S., Capel F. A monoclonal antibody specific for the hepatocyte receptor binding site on hepatitis B virus. Mol Immunol. 1989 Jun;26(6):531–537. doi: 10.1016/0161-5890(89)90004-7. [DOI] [PubMed] [Google Scholar]
  31. Ponzetto A., Cote P. J., Popper H., Hoyer B. H., London W. T., Ford E. C., Bonino F., Purcell R. H., Gerin J. L. Transmission of the hepatitis B virus-associated delta agent to the eastern woodchuck. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2208–2212. doi: 10.1073/pnas.81.7.2208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Riesner D., Gross H. J. Viroids. Annu Rev Biochem. 1985;54:531–564. doi: 10.1146/annurev.bi.54.070185.002531. [DOI] [PubMed] [Google Scholar]
  33. Rizzetto M., Hoyer B., Canese M. G., Shih J. W., Purcell R. H., Gerin J. L. delta Agent: association of delta antigen with hepatitis B surface antigen and RNA in serum of delta-infected chimpanzees. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6124–6128. doi: 10.1073/pnas.77.10.6124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sureau C., Jacob J. R., Eichberg J. W., Lanford R. E. Tissue culture system for infection with human hepatitis delta virus. J Virol. 1991 Jul;65(7):3443–3450. doi: 10.1128/jvi.65.7.3443-3450.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sureau C., Romet-Lemonne J. L., Mullins J. I., Essex M. Production of hepatitis B virus by a differentiated human hepatoma cell line after transfection with cloned circular HBV DNA. Cell. 1986 Oct 10;47(1):37–47. doi: 10.1016/0092-8674(86)90364-8. [DOI] [PubMed] [Google Scholar]
  36. Sureau C., Taylor J., Chao M., Eichberg J. W., Lanford R. E. Cloned hepatitis delta virus cDNA is infectious in the chimpanzee. J Virol. 1989 Oct;63(10):4292–4297. doi: 10.1128/jvi.63.10.4292-4297.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Taylor J. M. Hepatitis delta virus: cis and trans functions required for replication. Cell. 1990 May 4;61(3):371–373. doi: 10.1016/0092-8674(90)90516-h. [DOI] [PubMed] [Google Scholar]
  38. Taylor J., Mason W., Summers J., Goldberg J., Aldrich C., Coates L., Gerin J., Gowans E. Replication of human hepatitis delta virus in primary cultures of woodchuck hepatocytes. J Virol. 1987 Sep;61(9):2891–2895. doi: 10.1128/jvi.61.9.2891-2895.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tiollais P., Pourcel C., Dejean A. The hepatitis B virus. Nature. 1985 Oct 10;317(6037):489–495. doi: 10.1038/317489a0. [DOI] [PubMed] [Google Scholar]
  40. Ueda K., Tsurimoto T., Matsubara K. Three envelope proteins of hepatitis B virus: large S, middle S, and major S proteins needed for the formation of Dane particles. J Virol. 1991 Jul;65(7):3521–3529. doi: 10.1128/jvi.65.7.3521-3529.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wang K. S., Choo Q. L., Weiner A. J., Ou J. H., Najarian R. C., Thayer R. M., Mullenbach G. T., Denniston K. J., Gerin J. L., Houghton M. Structure, sequence and expression of the hepatitis delta (delta) viral genome. Nature. 1986 Oct 9;323(6088):508–514. doi: 10.1038/323508a0. [DOI] [PubMed] [Google Scholar]
  42. Weiner A. J., Choo Q. L., Wang K. S., Govindarajan S., Redeker A. G., Gerin J. L., Houghton M. A single antigenomic open reading frame of the hepatitis delta virus encodes the epitope(s) of both hepatitis delta antigen polypeptides p24 delta and p27 delta. J Virol. 1988 Feb;62(2):594–599. doi: 10.1128/jvi.62.2.594-599.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wu H. N., Lai M. M. Reversible cleavage and ligation of hepatitis delta virus RNA. Science. 1989 Feb 3;243(4891):652–654. doi: 10.1126/science.2492677. [DOI] [PubMed] [Google Scholar]
  44. Wu J. C., Chen P. J., Kuo M. Y., Lee S. D., Chen D. S., Ting L. P. Production of hepatitis delta virus and suppression of helper hepatitis B virus in a human hepatoma cell line. J Virol. 1991 Mar;65(3):1099–1104. doi: 10.1128/jvi.65.3.1099-1104.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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