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Archives of Virology

, Volume 161, Issue 3, pp 583–594 | Cite as

Replication of a chronic hepatitis B virus genotype F1b construct

  • Sergio Hernández
  • Gustavo Jiménez
  • Valentina Alarcón
  • Cristian Prieto
  • Francisca Muñoz
  • Constanza Riquelme
  • Mauricio Venegas
  • Javier Brahm
  • Alejandra Loyola
  • Rodrigo A VillanuevaEmail author
Original Article

Abstract

Genotype F is one of the less-studied genotypes of human hepatitis B virus, although it is widely distributed in regions of Central and South American. Our previous studies have shown that HBV genotype F is prevalent in Chile, and phylogenetic analysis of its full-length sequence amplified from the sera of chronically infected patients identified it as HBV subgenotype F1b. We have previously reported the full-length sequence of a HBV molecular clone obtained from a patient chronically infected with genotype F1b. In this report, we established a system to study HBV replication based on hepatoma cell lines transfected with full-length monomers of the HBV genome. Culture supernatants were analyzed after transfection and found to contain both HBsAg and HBeAg viral antigens. Consistently, fractionated cell extracts revealed the presence of viral replication, with both cytoplasmic and nuclear DNA intermediates. Analysis of HBV-transfected cells by indirect immunofluorescence or immunoelectron microscopy revealed the expression of viral antigens and cytoplasmic viral particles, respectively. To test the functionality of the ongoing viral replication further at the level of chromatinized cccDNA, transfected cells were treated with a histone deacetylase inhibitor, and this resulted in increased viral replication. This correlated with changes posttranslational modifications of histones at viral promoters. Thus, the development of this viral replication system for HBV genotype F will facilitate studies on the regulation of viral replication and the identification of new antiviral drugs.

Keywords

HepG2 Cell Sodium Butyrate Sodium Butyrate Treatment Viral cccDNA PreS1 Promoter 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This research was supported by grants from PIA-ANILLOS # ACT1119, FONDECYT # 1120170, Basal Project # PFB16, and Universidad San Sebastián 2011-005-R, PCHA/Doctorado Nacional/2014-21140956 (SH), 2012-21120216 (FM).

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Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Sergio Hernández
    • 1
  • Gustavo Jiménez
    • 1
  • Valentina Alarcón
    • 2
  • Cristian Prieto
    • 1
  • Francisca Muñoz
    • 2
  • Constanza Riquelme
    • 1
  • Mauricio Venegas
    • 3
  • Javier Brahm
    • 3
  • Alejandra Loyola
    • 2
    • 4
  • Rodrigo A Villanueva
    • 1
    Email author
  1. 1.Laboratorio de Virus Hepatitis, Facultad de Ciencias BiológicasUniversidad Andrés BelloSantiagoChile
  2. 2.Laboratorio de Epigenética y Cromatina, Fundación Ciencia and VidaSantiagoChile
  3. 3.Sección de GastroenterologíaHospital Clínico Universidad de ChileSantiagoChile
  4. 4.Universidad San SebastiánSantiagoChile

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