1887

Abstract

The aim of this study was to investigate the infection and replication of swine-derived hepatitis E virus (HEV) in primary cultured human hepatocytes (PHCs). Hepatocytes were cultured from the resected normal livers of patients with metastatic tumours. These cultured hepatocytes were infected with swine-derived genotype 3 or 4 HEV. Viral replication was monitored using reverse transcriptase-quantitative PCR. The amount of HEV RNA increased in the culture media and cells following infection. Immunofluorescence staining implied that the spread of HEV infection in hepatocytes was attributed mainly to cell-to-cell transmission via the cell membrane. The sequences of the inoculated and propagated HEV were determined to examine whether sequence variation occurred during infection. Sequence analysis showed that there were no differences between inoculated and propagated HEV, demonstrating that infection and replication of swine HEV in PHCs occurred without sequence variation.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.068452-0
2014-12-01
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/jgv/95/12/2677.html?itemId=/content/journal/jgv/10.1099/vir.0.068452-0&mimeType=html&fmt=ahah

References

  1. Datta R., Panda S. K., Tandon B. N., Madnagopalan N., Bose S. L., Acharya S. K., Nayak N. C. 1987; Acute sporadic non-A, non-B viral hepatitis of adults in India – epidemiological and immunological studies. J Gastroenterol Hepatol 2:333–345 [View Article]
    [Google Scholar]
  2. Emerson S. U., Nguyen H., Graff J., Stephany D. A., Brockington A., Purcell R. H. 2004a; In vitro replication of hepatitis E virus (HEV) genomes and of an HEV replicon expressing green fluorescent protein. J Virol 78:4838–4846 [View Article][PubMed]
    [Google Scholar]
  3. Emerson S. U., Anderson D., Arankalle A., Meng X. J., Purdy M., Schlauder G. G., Tsarev S. A. 2004b; Hepevirus. In Virus Taxonomy: VIIIth Report of the International Committee on Taxonomy of Viruses pp. 851–855 Edited by Fauquet C. M., Mayo M. A., Maniloff J., Desselberger U., Ball L. A. London: Elsevier;
    [Google Scholar]
  4. Guguen-Guillouzo C. 1992; Isolation and culture of animal and human hepatocytes. In Culture of Epithelial Cells pp. 197–223 Edited by Freshney R. I. New York: Wiley-Liss;
    [Google Scholar]
  5. Huang R., Li D., Wei S., Li Q., Yuan X., Geng L., Li X., Liu M. 1999; Cell culture of sporadic hepatitis E virus in China. Clin Diagn Lab Immunol 6:729–733[PubMed]
    [Google Scholar]
  6. Huang Y. W., Haqshenas G., Kasorndorkbua C., Halbur P. G., Emerson S. U., Meng X. J. 2005; Capped RNA transcripts of full-length cDNA clones of swine hepatitis E virus are replication competent when transfected into Huh7 cells and infectious when intrahepatically inoculated into pigs. J Virol 79:1552–1558 [View Article][PubMed]
    [Google Scholar]
  7. Jothikumar N., Cromeans T. L., Robertson B. H., Meng X. J., Hill V. R. 2006; A broadly reactive one-step real-time RT-PCR assay for rapid and sensitive detection of hepatitis E virus. J Virol Methods 131:65–71 [View Article][PubMed]
    [Google Scholar]
  8. Kamar N., Selves J., Mansuy J. M., Ouezzani L., Péron J. M., Guitard J., Cointault O., Esposito L., Abravanel F. other authors 2008; Hepatitis E virus and chronic hepatitis in organ-transplant recipients. N Engl J Med 358:811–817 [View Article][PubMed]
    [Google Scholar]
  9. Kamar N., Bendall R., Legrand-Abravanel F., Xia N. S., Ijaz S., Izopet J., Dalton H. R. 2012; Hepatitis E. Lancet 379:2477–2488 [View Article][PubMed]
    [Google Scholar]
  10. Li T. C., Yamakawa Y., Suzuki K., Tatsumi M., Razak M. A., Uchida T., Takeda N., Miyamura T. 1997; Expression and self-assembly of empty virus-like particles of hepatitis E virus. J Virol 71:7207–7213[PubMed]
    [Google Scholar]
  11. Li W., Sun Q., She R., Wang D., Duan X., Yin J., Ding Y. 2009; Experimental infection of Mongolian gerbils by a genotype 4 strain of swine hepatitis E virus. J Med Virol 81:1591–1596 [View Article][PubMed]
    [Google Scholar]
  12. Lorenzo F. R., Tanaka T., Takahashi H., Ichiyama K., Hoshino Y., Yamada K., Inoue J., Takahashi M., Okamoto H. 2008; Mutational events during the primary propagation and consecutive passages of hepatitis E virus strain JE03-1760F in cell culture. Virus Res 137:86–96 [View Article][PubMed]
    [Google Scholar]
  13. Lu L., Li C., Hagedorn C. H. 2006; Phylogenetic analysis of global hepatitis E virus sequences: genetic diversity, subtypes and zoonosis. Rev Med Virol 16:5–36 [View Article][PubMed]
    [Google Scholar]
  14. Ma H., Zheng L., Liu Y., Zhao C., Harrison T. J., Ma Y., Sun S., Zhang J., Wang Y. 2010; Experimental infection of rabbits with rabbit and genotypes 1 and 4 hepatitis E viruses. PLoS ONE 5:e9160 [View Article][PubMed]
    [Google Scholar]
  15. Meng X. J., Purcell R. H., Halbur P. G., Lehman J. R., Webb D. M., Tsareva T. S., Haynes J. S., Thacker B. J., Emerson S. U. 1997; A novel virus in swine is closely related to the human hepatitis E virus. Proc Natl Acad Sci U S A 94:9860–9865 [View Article][PubMed]
    [Google Scholar]
  16. Okamoto H., Takahashi M., Nishizawa T. 2003; Features of hepatitis E virus infection in Japan. Intern Med 42:1065–1071 [View Article][PubMed]
    [Google Scholar]
  17. Rumin S., Gripon P., Le Seyec J., Corral-Debrinski M., Guguen-Guillouzo C. 1996; Long-term productive episomal hepatitis B virus replication in primary cultures of adult human hepatocytes infected in vitro . J Viral Hepat 3:227–238 [View Article][PubMed]
    [Google Scholar]
  18. Shukla P., Nguyen H. T., Torian U., Engle R. E., Faulk K., Dalton H. R., Bendall R. P., Keane F. E., Purcell R. H., Emerson S. U. 2011; Cross-species infections of cultured cells by hepatitis E virus and discovery of an infectious virus–host recombinant. Proc Natl Acad Sci U S A 108:2438–2443 [View Article][PubMed]
    [Google Scholar]
  19. Takahashi M., Nishizawa T., Miyajima H., Gotanda Y., Iita T., Tsuda F., Okamoto H. 2003; Swine hepatitis E virus strains in Japan form four phylogenetic clusters comparable with those of Japanese isolates of human hepatitis E virus. J Gen Virol 84:851–862 [View Article][PubMed]
    [Google Scholar]
  20. Takahashi H., Tanaka T., Jirintai S., Nagashima S., Takahashi M., Nishizawa T., Mizuo H., Yazaki Y., Okamoto H. 2012; A549 and PLC/PRF/5 cells can support the efficient propagation of swine and wild boar hepatitis E virus (HEV) strains: demonstration of HEV infectivity of porcine liver sold as food. Arch Virol 157:235–246 [View Article][PubMed]
    [Google Scholar]
  21. Tanaka T., Takahashi M., Kusano E., Okamoto H. 2007; Development and evaluation of an efficient cell-culture system for hepatitis E virus. J Gen Virol 88:903–911 [View Article][PubMed]
    [Google Scholar]
  22. Tei S., Kitajima N., Takahashi K., Mishiro S. 2003; Zoonotic transmission of hepatitis E virus from deer to human beings. Lancet 362:371–373 [View Article][PubMed]
    [Google Scholar]
  23. Urayama T., Sapsutthipas S., Tsujikawa M., Yamashita A., Nishigaki H., Ibrahim M. S., Hagiwara K., Yunoki M., Yasunaga T. other authors 2010; Full-length sequences of one genotype 4 and three genotype 3 hepatitis E viruses in fecal samples from domestic swine in Japan. Open Vet Sci J 4:11–19 [View Article]
    [Google Scholar]
  24. Weiss T. S., Jahn B., Cetto M., Jauch K. W., Thasler W. E. 2002; Collagen sandwich culture affects intracellular polyamine levels of human hepatocytes. Cell Prolif 35:257–267 [View Article][PubMed]
    [Google Scholar]
  25. Yamada K., Takahashi M., Hoshino Y., Takahashi H., Ichiyama K., Tanaka T., Okamoto H. 2009; Construction of an infectious cDNA clone of hepatitis E virus strain JE03-1760F that can propagate efficiently in cultured cells. J Gen Virol 90:457–462 [CrossRef]
    [Google Scholar]
  26. Yunoki M., Yamamoto S., Tanaka H., Nishigaki H., Tanaka Y., Nishida A., Adan-Kubo J., Tsujikawa M., Hattori S. other authors 2008; Extent of hepatitis E virus elimination is affected by stabilizers present in plasma products and pore size of nanofilters. Vox Sang 95:94–100 [View Article][PubMed]
    [Google Scholar]
  27. Zhai L., Dai X., Meng J. 2006; Hepatitis E virus genotyping based on full-length genome and partial genomic regions. Virus Res 120:57–69 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.068452-0
Loading
/content/journal/jgv/10.1099/vir.0.068452-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error