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. 1995 Oct;69(10):6158–6169. doi: 10.1128/jvi.69.10.6158-6169.1995

High-level hepatitis B virus replication in transgenic mice.

L G Guidotti 1, B Matzke 1, H Schaller 1, F V Chisari 1
PMCID: PMC189513  PMID: 7666518

Abstract

Hepatitis B virus (HBV) transgenic mice whose hepatocytes replicate the virus at levels comparable to that in the infected livers of patients with chronic hepatitis have been produced, without any evidence of cytopathology. High-level viral gene expression was obtained in the liver and kidney tissues in three independent lineages. These animals were produced with a terminally redundant viral DNA construct (HBV 1.3) that starts just upstream of HBV enhancer I, extends completely around the circular viral genome, and ends just downstream of the unique polyadenylation site in HBV. In these animals, the viral mRNA is more abundant in centrilobular hepatocytes than elsewhere in the hepatic lobule. High-level viral DNA replication occurs inside viral nucleocapsid particles that preferentially form in the cytoplasm of these centrilobular hepatocytes, suggesting that an expression threshold must be reached for nucleocapsid assembly and viral replication to occur. Despite the restricted distribution of the viral replication machinery in centrilobular cytoplasmic nucleocapsids, nucleocapsid particles are detectable in the vast majority of hepatocyte nuclei throughout the hepatic lobule. The intranuclear nucleocapsid particles are empty, however, suggesting that viral nucleocapsid particle assembly occurs independently in the nucleus and the cytoplasm of the hepatocyte and implying that cytoplasmic nucleocapsid particles do not transport the viral genome across the nuclear membrane into the nucleus during the viral life cycle. This model creates the opportunity to examine the influence of viral and host factors on HBV pathogenesis and replication and to assess the antiviral potential of pharmacological agents and physiological processes, including the immune response.

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Selected References

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  1. Araki K., Miyazaki J., Hino O., Tomita N., Chisaka O., Matsubara K., Yamamura K. Expression and replication of hepatitis B virus genome in transgenic mice. Proc Natl Acad Sci U S A. 1989 Jan;86(1):207–211. doi: 10.1073/pnas.86.1.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Babinet C., Farza H., Morello D., Hadchouel M., Pourcel C. Specific expression of hepatitis B surface antigen (HBsAg) in transgenic mice. Science. 1985 Dec 6;230(4730):1160–1163. doi: 10.1126/science.3865370. [DOI] [PubMed] [Google Scholar]
  3. Burk R. D., DeLoia J. A., elAwady M. K., Gearhart J. D. Tissue preferential expression of the hepatitis B virus (HBV) surface antigen gene in two lines of HBV transgenic mice. J Virol. 1988 Feb;62(2):649–654. doi: 10.1128/jvi.62.2.649-654.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Calvert J., Summers J. Two regions of an avian hepadnavirus RNA pregenome are required in cis for encapsidation. J Virol. 1994 Apr;68(4):2084–2090. doi: 10.1128/jvi.68.4.2084-2090.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chisari F. V., Ferrari C. Hepatitis B virus immunopathogenesis. Annu Rev Immunol. 1995;13:29–60. doi: 10.1146/annurev.iy.13.040195.000333. [DOI] [PubMed] [Google Scholar]
  6. Chisari F. V., Filippi P., McLachlan A., Milich D. R., Riggs M., Lee S., Palmiter R. D., Pinkert C. A., Brinster R. L. Expression of hepatitis B virus large envelope polypeptide inhibits hepatitis B surface antigen secretion in transgenic mice. J Virol. 1986 Dec;60(3):880–887. doi: 10.1128/jvi.60.3.880-887.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chisari F. V., Pinkert C. A., Milich D. R., Filippi P., McLachlan A., Palmiter R. D., Brinster R. L. A transgenic mouse model of the chronic hepatitis B surface antigen carrier state. Science. 1985 Dec 6;230(4730):1157–1160. doi: 10.1126/science.3865369. [DOI] [PubMed] [Google Scholar]
  8. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  9. Farza H., Hadchouel M., Scotto J., Tiollais P., Babinet C., Pourcel C. Replication and gene expression of hepatitis B virus in a transgenic mouse that contains the complete viral genome. J Virol. 1988 Nov;62(11):4144–4152. doi: 10.1128/jvi.62.11.4144-4152.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Gilles P. N., Fey G., Chisari F. V. Tumor necrosis factor alpha negatively regulates hepatitis B virus gene expression in transgenic mice. J Virol. 1992 Jun;66(6):3955–3960. doi: 10.1128/jvi.66.6.3955-3960.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guidotti L. G., Ando K., Hobbs M. V., Ishikawa T., Runkel L., Schreiber R. D., Chisari F. V. Cytotoxic T lymphocytes inhibit hepatitis B virus gene expression by a noncytolytic mechanism in transgenic mice. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3764–3768. doi: 10.1073/pnas.91.9.3764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Guidotti L. G., Martinez V., Loh Y. T., Rogler C. E., Chisari F. V. Hepatitis B virus nucleocapsid particles do not cross the hepatocyte nuclear membrane in transgenic mice. J Virol. 1994 Sep;68(9):5469–5475. doi: 10.1128/jvi.68.9.5469-5475.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Guilhot S., Fowler P., Portillo G., Margolskee R. F., Ferrari C., Bertoletti A., Chisari F. V. Hepatitis B virus (HBV)-specific cytotoxic T-cell response in humans: production of target cells by stable expression of HBV-encoded proteins in immortalized human B-cell lines. J Virol. 1992 May;66(5):2670–2678. doi: 10.1128/jvi.66.5.2670-2678.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Guilhot S., Guidotti L. G., Chisari F. V. Interleukin-2 downregulates hepatitis B virus gene expression in transgenic mice by a posttranscriptional mechanism. J Virol. 1993 Dec;67(12):7444–7449. doi: 10.1128/jvi.67.12.7444-7449.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hoofnagle J. H., Shafritz D. A., Popper H. Chronic type B hepatitis and the "healthy" HBsAg carrier state. Hepatology. 1987 Jul-Aug;7(4):758–763. doi: 10.1002/hep.1840070424. [DOI] [PubMed] [Google Scholar]
  17. Junker-Niepmann M., Bartenschlager R., Schaller H. A short cis-acting sequence is required for hepatitis B virus pregenome encapsidation and sufficient for packaging of foreign RNA. EMBO J. 1990 Oct;9(10):3389–3396. doi: 10.1002/j.1460-2075.1990.tb07540.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kajino K., Jilbert A. R., Saputelli J., Aldrich C. E., Cullen J., Mason W. S. Woodchuck hepatitis virus infections: very rapid recovery after a prolonged viremia and infection of virtually every hepatocyte. J Virol. 1994 Sep;68(9):5792–5803. doi: 10.1128/jvi.68.9.5792-5803.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Katayama K., Hayashi N., Sasaki Y., Kasahara A., Ueda K., Fusamoto H., Sato N., Chisaka O., Matsubara K., Kamada T. Detection of hepatitis B virus X gene protein and antibody in type B chronic liver disease. Gastroenterology. 1989 Oct;97(4):990–998. doi: 10.1016/0016-5085(89)91508-4. [DOI] [PubMed] [Google Scholar]
  20. Kim C. M., Koike K., Saito I., Miyamura T., Jay G. HBx gene of hepatitis B virus induces liver cancer in transgenic mice. Nature. 1991 May 23;351(6324):317–320. doi: 10.1038/351317a0. [DOI] [PubMed] [Google Scholar]
  21. Köck J., Schlicht H. J. Analysis of the earliest steps of hepadnavirus replication: genome repair after infectious entry into hepatocytes does not depend on viral polymerase activity. J Virol. 1993 Aug;67(8):4867–4874. doi: 10.1128/jvi.67.8.4867-4874.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lee T. H., Finegold M. J., Shen R. F., DeMayo J. L., Woo S. L., Butel J. S. Hepatitis B virus transactivator X protein is not tumorigenic in transgenic mice. J Virol. 1990 Dec;64(12):5939–5947. doi: 10.1128/jvi.64.12.5939-5947.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McLachlan A., Milich D. R., Raney A. K., Riggs M. G., Hughes J. L., Sorge J., Chisari F. V. Expression of hepatitis B virus surface and core antigens: influences of pre-S and precore sequences. J Virol. 1987 Mar;61(3):683–692. doi: 10.1128/jvi.61.3.683-692.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Milich D. R., Jones J. E., Hughes J. L., Maruyama T., Price J., Melhado I., Jirik F. Extrathymic expression of the intracellular hepatitis B core antigen results in T cell tolerance in transgenic mice. J Immunol. 1994 Jan 15;152(2):455–466. [PubMed] [Google Scholar]
  25. Milich D. R., Jones J. E., Hughes J. L., Price J., Raney A. K., McLachlan A. Is a function of the secreted hepatitis B e antigen to induce immunologic tolerance in utero? Proc Natl Acad Sci U S A. 1990 Sep;87(17):6599–6603. doi: 10.1073/pnas.87.17.6599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Radziwill G., Tucker W., Schaller H. Mutational analysis of the hepatitis B virus P gene product: domain structure and RNase H activity. J Virol. 1990 Feb;64(2):613–620. doi: 10.1128/jvi.64.2.613-620.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ranki M., Schätzl H. M., Zachoval R., Uusi-Oukari M., Lehtovaara P. Quantification of hepatitis B virus DNA over a wide range from serum for studying viral replicative activity in response to treatment and in recurrent infection. Hepatology. 1995 Jun;21(6):1492–1499. [PubMed] [Google Scholar]
  28. Sladek F. M., Darnell J. E. Mechanisms of liver-specific gene expression. Curr Opin Genet Dev. 1992 Apr;2(2):256–259. doi: 10.1016/s0959-437x(05)80282-5. [DOI] [PubMed] [Google Scholar]
  29. Summers J., Mason W. S. Replication of the genome of a hepatitis B--like virus by reverse transcription of an RNA intermediate. Cell. 1982 Jun;29(2):403–415. doi: 10.1016/0092-8674(82)90157-x. [DOI] [PubMed] [Google Scholar]
  30. Summers J., Smith P. M., Horwich A. L. Hepadnavirus envelope proteins regulate covalently closed circular DNA amplification. J Virol. 1990 Jun;64(6):2819–2824. doi: 10.1128/jvi.64.6.2819-2824.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Treinin M., Laub O. Identification of a promoter element located upstream from the hepatitis B virus X gene. Mol Cell Biol. 1987 Jan;7(1):545–548. doi: 10.1128/mcb.7.1.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tuttleman J. S., Pourcel C., Summers J. Formation of the pool of covalently closed circular viral DNA in hepadnavirus-infected cells. Cell. 1986 Nov 7;47(3):451–460. doi: 10.1016/0092-8674(86)90602-1. [DOI] [PubMed] [Google Scholar]
  33. Tuttleman J. S., Pugh J. C., Summers J. W. In vitro experimental infection of primary duck hepatocyte cultures with duck hepatitis B virus. J Virol. 1986 Apr;58(1):17–25. doi: 10.1128/jvi.58.1.17-25.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Will H., Cattaneo R., Darai G., Deinhardt F., Schellekens H., Schaller H. Infectious hepatitis B virus from cloned DNA of known nucleotide sequence. Proc Natl Acad Sci U S A. 1985 Feb;82(3):891–895. doi: 10.1073/pnas.82.3.891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zoulim F., Saputelli J., Seeger C. Woodchuck hepatitis virus X protein is required for viral infection in vivo. J Virol. 1994 Mar;68(3):2026–2030. doi: 10.1128/jvi.68.3.2026-2030.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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