Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1992 Nov;66(11):6649–6654. doi: 10.1128/jvi.66.11.6649-6654.1992

cDNA clone of hepatitis A virus encoding a virulent virus: induction of viral hepatitis by direct nucleic acid transfection of marmosets.

S U Emerson 1, M Lewis 1, S Govindarajan 1, M Shapiro 1, T Moskal 1, R H Purcell 1
PMCID: PMC240161  PMID: 1328684

Abstract

Direct inoculation of marmoset livers with an in vitro transcription mixture containing cDNA and full-length genomic RNA transcripts of hepatitis A virus resulted in acute viral hepatitis. Elevations in serum levels of liver enzymes were correlated with appearance of antibody to hepatitis A virus. Genomes of infectious hepatitis A virus isolated from the feces of transfected marmosets contained the same mutation as the cDNA template used for transfection. Liver biopsies confirmed that the virus encoded by the cDNA clone induced histopathological changes equivalent to those caused by virulent wild-type virus.

Full text

PDF
6649

Images in this article

Selected References

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

  1. Calenoff M. A., Faaberg K. S., Lipton H. L. Genomic regions of neurovirulence and attenuation in Theiler murine encephalomyelitis virus. Proc Natl Acad Sci U S A. 1990 Feb;87(3):978–982. doi: 10.1073/pnas.87.3.978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Cohen J. I., Rosenblum B., Feinstone S. M., Ticehurst J., Purcell R. H. Attenuation and cell culture adaptation of hepatitis A virus (HAV): a genetic analysis with HAV cDNA. J Virol. 1989 Dec;63(12):5364–5370. doi: 10.1128/jvi.63.12.5364-5370.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cohen J. I., Rosenblum B., Ticehurst J. R., Daemer R. J., Feinstone S. M., Purcell R. H. Complete nucleotide sequence of an attenuated hepatitis A virus: comparison with wild-type virus. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2497–2501. doi: 10.1073/pnas.84.8.2497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen J. I., Ticehurst J. R., Feinstone S. M., Rosenblum B., Purcell R. H. Hepatitis A virus cDNA and its RNA transcripts are infectious in cell culture. J Virol. 1987 Oct;61(10):3035–3039. doi: 10.1128/jvi.61.10.3035-3039.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Daemer R. J., Feinstone S. M., Gust I. D., Purcell R. H. Propagation of human hepatitis A virus in African green monkey kidney cell culture: primary isolation and serial passage. Infect Immun. 1981 Apr;32(1):388–393. doi: 10.1128/iai.32.1.388-393.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Emerson S. U., Huang Y. K., McRill C., Lewis M., Purcell R. H. Mutations in both the 2B and 2C genes of hepatitis A virus are involved in adaptation to growth in cell culture. J Virol. 1992 Feb;66(2):650–654. doi: 10.1128/jvi.66.2.650-654.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Emerson S. U., McRill C., Rosenblum B., Feinstone S., Purcell R. H. Mutations responsible for adaptation of hepatitis A virus to efficient growth in cell culture. J Virol. 1991 Sep;65(9):4882–4886. doi: 10.1128/jvi.65.9.4882-4886.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fu J. L., Stein S., Rosenstein L., Bodwell T., Routbort M., Semler B. L., Roos R. P. Neurovirulence determinants of genetically engineered Theiler viruses. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4125–4129. doi: 10.1073/pnas.87.11.4125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Karron R. A., Daemer R., Ticehurst J., D'Hondt E., Popper H., Mihalik K., Phillips J., Feinstone S., Purcell R. H. Studies of prototype live hepatitis A virus vaccines in primate models. J Infect Dis. 1988 Feb;157(2):338–345. doi: 10.1093/infdis/157.2.338. [DOI] [PubMed] [Google Scholar]
  11. Lemon S. M., Murphy P. C., Shields P. A., Ping L. H., Feinstone S. M., Cromeans T., Jansen R. W. Antigenic and genetic variation in cytopathic hepatitis A virus variants arising during persistent infection: evidence for genetic recombination. J Virol. 1991 Apr;65(4):2056–2065. doi: 10.1128/jvi.65.4.2056-2065.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ohlinger V. F., Haas B., Meyers G., Weiland F., Thiel H. J. Identification and characterization of the virus causing rabbit hemorrhagic disease. J Virol. 1990 Jul;64(7):3331–3336. doi: 10.1128/jvi.64.7.3331-3336.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Paul A. V., Tada H., von der Helm K., Wissel T., Kiehn R., Wimmer E., Deinhardt F. The entire nucleotide sequence of the genome of human hepatitis A virus (isolate MBB). Virus Res. 1987 Aug;8(2):153–171. doi: 10.1016/0168-1702(87)90026-8. [DOI] [PubMed] [Google Scholar]
  14. Ren R. B., Moss E. G., Racaniello V. R. Identification of two determinants that attenuate vaccine-related type 2 poliovirus. J Virol. 1991 Mar;65(3):1377–1382. doi: 10.1128/jvi.65.3.1377-1382.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ross B. C., Anderson B. N., Edwards P. C., Gust I. D. Nucleotide sequence of high-passage hepatitis A virus strain HM175: comparison with wild-type and cell culture-adapted strains. J Gen Virol. 1989 Oct;70(Pt 10):2805–2810. doi: 10.1099/0022-1317-70-10-2805. [DOI] [PubMed] [Google Scholar]
  16. Seeger C., Ganem D., Varmus H. E. The cloned genome of ground squirrel hepatitis virus is infectious in the animal. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5849–5852. doi: 10.1073/pnas.81.18.5849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Winship P. R. An improved method for directly sequencing PCR amplified material using dimethyl sulphoxide. Nucleic Acids Res. 1989 Feb 11;17(3):1266–1266. doi: 10.1093/nar/17.3.1266. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES