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. 1987 Jul;84(13):4641–4644. doi: 10.1073/pnas.84.13.4641

Hepatitis B virus produced by transfected Hep G2 cells causes hepatitis in chimpanzees.

G Acs, M A Sells, R H Purcell, P Price, R Engle, M Shapiro, H Popper
PMCID: PMC305146  PMID: 2885842

Abstract

We have reported that clonal cells derived from Hep G2 cells transfected with a plasmid containing hepatitis B virus (HBV) DNA secrete spherical and filamentous forms of hepatitis B surface antigen (HBsAg), core particles, and virions into the culture medium. Here we describe the development of typical hepatitis in two chimpanzees following intravenous inoculation with the medium in which the transfected cells had grown. The liver biopsies from these animals showed characteristic lesions in parenchyma and portal tracts, more conspicuous at an earlier time in the chimpanzee that had received a greater number of virions. The amount of HBsAg in the serum of one infected chimpanzee increased with time after the initial inoculation and then decreased concomitantly with the appearance of antibodies against HBsAg and core antigens. HBsAg remained detectable in the other animal throughout the course of the experiment. The levels of hepatitis B "e" antigen in both animals peaked at week 5, signifying the acute phase of the infection. The activities of serum enzymes that are markers for necroinflammation also increased. The hepatitis HBsAg subtype of the virions isolated from the patient whose DNA was cloned and then used for transfection of the Hep G2 cells was the same as that found in the chimpanzees. Furthermore, the restriction enzyme analysis of the viral DNA isolated from the chimpanzees was identical to the cloned DNA. Thus, HBV DNA-transfected Hep G2 cells can support the replication of virions that, in turn, produce hepatitis in chimpanzees.

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Selected References

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

  1. Barker L. F., Chisari F. V., McGrath P. P., Dalgard D. W., Kirschstein R. L., Almeida J. D., Edington T. S., Sharp D. G., Peterson M. R. Transmission of type B viral hepatitis to chimpanzees. J Infect Dis. 1973 Jun;127(6):648–662. doi: 10.1093/infdis/127.6.648. [DOI] [PubMed] [Google Scholar]
  2. Barker L. F., Maynard J. E., Purcell R. H., Hoofnagle J. H., Berquist K. R., London W. T., Gerety R. J., Krushak D. H. Hepatitis B virus infection in chimpanzees: titration of subtypes. J Infect Dis. 1975 Oct;132(4):451–458. doi: 10.1093/infdis/132.4.451. [DOI] [PubMed] [Google Scholar]
  3. Berninger M., Hammer M., Hoyer B., Gerin J. L. An assay for the detection of the DNA genome of hepatitis B virus in serum. J Med Virol. 1982;9(1):57–68. doi: 10.1002/jmv.1890090109. [DOI] [PubMed] [Google Scholar]
  4. Dienstag J. L., Popper H., Purcell R. H. The pathology of viral hepatitis types A and B in chimpanzees. A comparison. Am J Pathol. 1976 Oct;85(1):131–148. [PMC free article] [PubMed] [Google Scholar]
  5. Galibert F., Mandart E., Fitoussi F., Tiollais P., Charnay P. Nucleotide sequence of the hepatitis B virus genome (subtype ayw) cloned in E. coli. Nature. 1979 Oct 25;281(5733):646–650. doi: 10.1038/281646a0. [DOI] [PubMed] [Google Scholar]
  6. Hoofnagle J. H., Gerety R. J., Smallwood L. A., Barker L. F. Subtyping of hepatitis B surface antigen and antibody by radioimmunoassay. Gastroenterology. 1977 Feb;72(2):290–296. [PubMed] [Google Scholar]
  7. Price P., Hirschman S. Z., Garfinkel E. DNA cloned from the ayw subtype of hepatitis B virus. J Med Virol. 1980;6(2):139–145. doi: 10.1002/jmv.1890060206. [DOI] [PubMed] [Google Scholar]
  8. Reed K. C., Mann D. A. Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 1985 Oct 25;13(20):7207–7221. doi: 10.1093/nar/13.20.7207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Sells M. A., Chen M. L., Acs G. Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc Natl Acad Sci U S A. 1987 Feb;84(4):1005–1009. doi: 10.1073/pnas.84.4.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Tsurimoto T., Fujiyama A., Matsubara K. Stable expression and replication of hepatitis B virus genome in an integrated state in a human hepatoma cell line transfected with the cloned viral DNA. Proc Natl Acad Sci U S A. 1987 Jan;84(2):444–448. doi: 10.1073/pnas.84.2.444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Will H., Cattaneo R., Koch H. G., Darai G., Schaller H., Schellekens H., van Eerd P. M., Deinhardt F. Cloned HBV DNA causes hepatitis in chimpanzees. Nature. 1982 Oct 21;299(5885):740–742. doi: 10.1038/299740a0. [DOI] [PubMed] [Google Scholar]

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