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. Villanueva

Resultado de la investigación: Article

1 Cita (Scopus)

Resumen

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.

Idioma originalEnglish
Páginas (desde-hasta)583-594
Número de páginas12
PublicaciónArchives of Virology
Volumen161
N.º3
DOI
EstadoPublished - 1 mar 2016

Huella dactilar

Chronic Hepatitis B
Hepatitis B virus
Genotype
Viral Antigens
Histone Deacetylase Inhibitors
Hepatitis B e Antigens
Immunoelectron Microscopy
Chile
Post Translational Protein Processing
Indirect Fluorescent Antibody Technique
Hepatitis B Surface Antigens
Cell Extracts
Fluorescence Microscopy
Virion
Histones
Antiviral Agents
Transfection
Hepatocellular Carcinoma
Clone Cells
Genome

ASJC Scopus subject areas

  • Virology

Citar esto

Hernández, S., Jiménez, G., Alarcón, V., Prieto, C., Muñoz, F., Riquelme, C., ... Villanueva, R. A. (2016). Replication of a chronic hepatitis B virus genotype F1b construct. Archives of Virology, 161(3), 583-594. https://doi.org/10.1007/s00705-015-2702-x
Hernández, Sergio ; Jiménez, Gustavo ; Alarcón, Valentina ; Prieto, Cristian ; Muñoz, Francisca ; Riquelme, Constanza ; Venegas, Mauricio ; Brahm, Javier ; Loyola, Alejandra ; Villanueva, Rodrigo A. / Replication of a chronic hepatitis B virus genotype F1b construct. En: Archives of Virology. 2016 ; Vol. 161, N.º 3. pp. 583-594.
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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.",
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Hernández, S, Jiménez, G, Alarcón, V, Prieto, C, Muñoz, F, Riquelme, C, Venegas, M, Brahm, J, Loyola, A & Villanueva, RA 2016, 'Replication of a chronic hepatitis B virus genotype F1b construct', Archives of Virology, vol. 161, n.º 3, pp. 583-594. https://doi.org/10.1007/s00705-015-2702-x

Replication of a chronic hepatitis B virus genotype F1b construct. / Hernández, Sergio; Jiménez, Gustavo; Alarcón, Valentina; Prieto, Cristian; Muñoz, Francisca; Riquelme, Constanza; Venegas, Mauricio; Brahm, Javier; Loyola, Alejandra; Villanueva, Rodrigo A.

En: Archives of Virology, Vol. 161, N.º 3, 01.03.2016, p. 583-594.

Resultado de la investigación: Article

TY - JOUR

T1 - Replication of a chronic hepatitis B virus genotype F1b construct

AU - Hernández, Sergio

AU - Jiménez, Gustavo

AU - Alarcón, Valentina

AU - Prieto, Cristian

AU - Muñoz, Francisca

AU - Riquelme, Constanza

AU - Venegas, Mauricio

AU - Brahm, Javier

AU - Loyola, Alejandra

AU - Villanueva, Rodrigo A.

PY - 2016/3/1

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N2 - 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.

AB - 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.

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Hernández S, Jiménez G, Alarcón V, Prieto C, Muñoz F, Riquelme C y otros. Replication of a chronic hepatitis B virus genotype F1b construct. Archives of Virology. 2016 mar 1;161(3):583-594. https://doi.org/10.1007/s00705-015-2702-x