title: Replication of a chronic hepatitis B virus genotype F1b construct.
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.
authors.affiliation: Laboratorio de Virus Hepatitis, Facultad de Ciencias Biológicas, Universidad Andrés Bello, Avda República 217, 2do piso, 8370146, Santiago, Chile.
authors.forename: Sergio
authors.initials: S
authors.lastname: Hernández
authors.affiliation: Laboratorio de Virus Hepatitis, Facultad de Ciencias Biológicas, Universidad Andrés Bello, Avda República 217, 2do piso, 8370146, Santiago, Chile.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Jiménez
authors.affiliation: Laboratorio de Epigenética y Cromatina, Fundación Ciencia and Vida, 7780272, Santiago, Chile.
authors.forename: Valentina
authors.initials: V
authors.lastname: Alarcón
authors.affiliation: Laboratorio de Virus Hepatitis, Facultad de Ciencias Biológicas, Universidad Andrés Bello, Avda República 217, 2do piso, 8370146, Santiago, Chile.
authors.forename: Cristian
authors.initials: C
authors.lastname: Prieto
authors.affiliation: Laboratorio de Epigenética y Cromatina, Fundación Ciencia and Vida, 7780272, Santiago, Chile.
authors.forename: Francisca
authors.initials: F
authors.lastname: Muñoz
authors.affiliation: Laboratorio de Virus Hepatitis, Facultad de Ciencias Biológicas, Universidad Andrés Bello, Avda República 217, 2do piso, 8370146, Santiago, Chile.
authors.forename: Constanza
authors.initials: C
authors.lastname: Riquelme
authors.affiliation: Sección de Gastroenterología, Hospital Clínico Universidad de Chile, 8380456, Santiago, Chile.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Venegas
authors.affiliation: Sección de Gastroenterología, Hospital Clínico Universidad de Chile, 8380456, Santiago, Chile.
authors.forename: Javier
authors.initials: J
authors.lastname: Brahm
authors.affiliation: Laboratorio de Epigenética y Cromatina, Fundación Ciencia and Vida, 7780272, Santiago, Chile.|Universidad San Sebastián, 7510157, Santiago, Chile.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Loyola
authors.affiliation: Laboratorio de Virus Hepatitis, Facultad de Ciencias Biológicas, Universidad Andrés Bello, Avda República 217, 2do piso, 8370146, Santiago, Chile. rodrigo.villanueva@unab.cl.
authors.forename: Rodrigo A
authors.identifier: http://orcid.org/0000-0003-4782-6865
authors.initials: RA
authors.lastname: Villanueva
mesh_terms: D002458:Cell Fractionation; D045744:Cell Line, Tumor; D003470:Culture Media / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis; D005838:Genotype; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006513:Hepatitis B e Antigens / Q000032:analysis; D006515:Hepatitis B virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D019694:Hepatitis B, Chronic / Q000821:virology; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D014779:Virus Replication
chemical_list: D003470:Culture Media; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens