Virus-liver cell interactions in duck hepatitis B virus infection

Jilbert, Allison R.; Freiman, John S.; Burrell, Christopher J.; Holmes, Mikaela; Gowans, Eric J.; Rowland, R.; Hall, Pauline; Cossart, Yvonne E.

doi:10.1016/0016-5085(88)90375-7

Cited by 45 publications

(38 citation statements)

References 11 publications

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“…10 11 hepatocytes, the mode of viral spread must therefore be very efficient. In vivo, clusters of virus-replicating cells are frequently observed during the early phases of infection (5). Moreover, similar clusters are also seen in primary hepatocyte cultures (unpublished data).…”

supporting

confidence: 54%

Spread of Hepatitis B Viruses In Vitro Requires Extracellular Progeny and May Be Codetermined by Polarized Egress

Funk

Hohenberg

Mhamdi

et al. 2004

J Virol

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Viruses can spread by different mechanisms: via intracellular particles through cell junctions to neighboring cells or via secreted virions to adjacent or remote cells. The observation of clusters of hepadnavirus-infected cells both in vivo and in primary hepatocytes neither proves the first mechanism nor excludes the second. In order to test which mechanism, if not both, is used by hepatitis B viruses in order to spread, we used primary duck hepatocytes and duck hepatitis B virus (DHBV) as an infection model. If extracellular progeny virus alone determines spreading, neutralizing antisera or drugs blocking virus binding to hepatocytes should abolish secondary infection. In order to test this, we used DHBV envelope-specific neutralizing antisera, as well as suramin, a known inhibitor of infection. Both reagents strongly reduced hepatocellular attachment of viral particles and almost completely abolished primary infection, whereas an ongoing intracellular infection was not affected as long as no progeny virus was released. In contrast, incubation of infected primary hepatocytes with these reagents during release of progeny virus completely prevented secondary infection. Moreover, the combination of electron and immunofluorescence microscopy analyses revealed the residence of viral particles in cytoplasmic vesicles preferentially located near the basolateral membrane of infected hepatocytes. Taken together, these data strongly suggest that hepatitis B viruses mainly spread by secreted, extracellular progeny and point to polarized egress of viral particles into intercellular compartments, which restricts their diffusion and favors transmission of virus to adjacent cells.

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supporting

confidence: 54%

Spread of Hepatitis B Viruses In Vitro Requires Extracellular Progeny and May Be Codetermined by Polarized Egress

Funk

Hohenberg

Mhamdi

et al. 2004

J Virol

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“…13 The titer of virus was determined by Southern blot hybridization following selective DNA extraction of enveloped virus particles, as previously described. 13,16,17 Animals. Day-old ducklings obtained from Metzer Farms (Redland, CA) were used for two independent experiments, experiment 1 and experiment 2.…”

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confidence: 99%

Acute Liver Injury Following Infection With A Cytopathic Strain of Duck Hepatitis B Virus

1999

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A variant avian hepadnavirus that has been shown to destroy hepatocytes in vitro was found to be cytopathic in vivo. A single amino acid change of glycine to glutamic acid at position 133 (G133E) in the preS protein of duck hepatitis B virus (DHBV) caused an increase in the intranuclear pool of viral covalently closed circular DNA (cccDNA), resulting in a transient elevation of viral replication and eventual hepatocyte destruction. In vivo viral infection with the G133E virus was compared with infection with wild-type virus over a 72-day period. Birds were inoculated with virus at day 2 post-hatch to ensure a high percentage of infected hepatocytes and potential persistence of virus. Birds infected with the G133E virus had increased periportal cellular proliferation and numerous lysed apoptotic hepatocytes following 100% infection of hepatocytes. The liver damage within G133E virus-infected birds subsided over time, resulting in mild chronic hepatitis that was similar to that observed within wild-type virusinfected birds. The subsidence of liver damage in G133E virus-infected birds coincided with a reduction of viral cccDNA to wild-type virus levels in the liver. Our study indicates that maintenance of wild-type levels of viral cccDNA promotes persistence of virus infection by establishing a noncytopathic infection. (HEPATOLOGY 1999;29:563-571.)Hepadnaviruses cause persistent productive infections of each infected hepatocyte. During the initiation of infection, the relaxed circular DNA (rcDNA) genome in the virion is converted to covalently closed circular DNA (cccDNA) in the nucleus. Viral cccDNA is used as the template for viral RNA transcription, resulting in the production of a viral RNA molecule (pregenome) that is greater than one genome in length. The RNA pregenome is encapsidated within a viral core particle, where it is reverse-transcribed by the viralencoded DNA polymerase to produce viral minus-strand DNA. 1 Plus-strand DNA synthesis is then carried out by the same viral DNA polymerase using the viral DNA minusstrand as a template. The final DNA product contained with the capsid is a double-stranded molecule that is held in a circular conformation by a short region of base-pairing between the 5Ј end of the two strands. This form of viral DNA is referred to as relaxed circular DNA, or rcDNA. The viral capsids that contain rcDNA associate with the viral envelope proteins at the endoplasmic reticulum and are secreted as virions. Alternatively, rcDNA molecules may enter into the nucleus of the infected hepatocyte, where they are converted to additional molecules of cccDNA and cause increased levels of viral replication. The production of new viral cccDNA is regulated by the viral large envelope protein (preS) by a mechanism that has not been biochemically defined. 2 Presumably, an interaction between the preS protein and rcDNAcontaining capsids is necessary for enveloped virus formation and promotes secretion of viral rcDNA from the hepatocyte as virions. Because defects in preS that specifically inhibit ...

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“…It is known that G133E-infected primary duck hepatocytes transiently produce cccDNA and virus at higher levels than do WT-infected cells (15) and therefore k g Ͼ k w . We set the rate constant for WT cccDNA synthesis equal to 1.6 day Ϫ1 (11,12) and varied k g at between 1.6 and 8.0 day…”

Section: ϫ3mentioning

confidence: 99%

Frequency of Spontaneous Mutations in an Avian Hepadnavirus Infection

Pult

Abbott

Zhang

et al. 2001

J Virol

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In this study, we measured the frequency of revertants of a cytopathic strain of the duck hepatitis B virus that bears a single nucleotide substitution in the pre-S envelope protein open reading frame, resulting in the amino acid substitution G133E. Cytopathic virus mixed with known amounts of a genetically marked wild-type virus was injected into ducklings. Virus outgrowth was accompanied by a coselection of wild-type and spontaneous revertants during recovery of the ducklings from the acute liver injury caused by death of the G133E-infected cells. The frequency of individual revertants in the selected noncytopathic virus population was estimated by determining the ratio of each revertant to the wild-type virus. Spontaneous revertants were found to be present at frequencies of 1 ؋ 10 ؊5 to 6 ؋ 10 ؊5 per G133E genome inoculated. A mathematical model was used to estimate that the mutation rate was 0.8 ؋ 10 ؊5 to 4.5 ؋ 10 ؊5 per nucleotide per generation.Duck hepatitis B virus (DHBV) belongs to the family Hepadnaviridae, a small group of enveloped viruses which cause persistent liver infections and replicate their DNA genomes through reverse transcription of an RNA intermediate (26). Because of the involvement of reverse transcription, the replication of the viral genome is assumed to be error prone and to give rise to a high degree of genetic variation. In humans, HBV can cause acute and chronic liver disease, and genetic variation is believed to be related to the clinical outcome of hepadnavirus infection (for a review, see reference 8). In recent years, it has become more fully appreciated how the presence of a complex mixture of variants in the virus population within a single host, the quasispecies, may enable rapid adaptation to changing environments (5), in particular to selection forces such as antiviral drugs (1), vaccines (4), immunomodulatory substances (20), and other changes in the host's immune response, leading to the emergence of various resistance and escape mutants (25).Major mechanisms contributing to the complexity of the viral quasispecies are the generation of errors during viral genome replication, environmentally induced mutations, and recombination. Of these, errors in replication are thought to constitute the greatest source for generating variants. The frequency of replication errors in hepadnaviruses is a function of the fidelities of three polymerization reactions which occur during the life cycle in two different compartments. In the nucleus, covalently closed circular DNA (cccDNA)-templated positive-strand RNA synthesis is carried out by cellular RNA polymerase (RNA pregenome synthesis), and in the viral core particles, RNA-templated negative-strand DNA synthesis and DNA-templated positive-strand DNA synthesis are both carried out by reverse transcriptase. The relative contributions of the various enzymes involved in different polymerization steps to the overall mutation rate of the virus are not known.In spite of the reported widespread occurrence of genetic variation in hepadnaviru...

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Virus-liver cell interactions in duck hepatitis B virus infection

Cited by 45 publications

References 11 publications

Spread of Hepatitis B Viruses In Vitro Requires Extracellular Progeny and May Be Codetermined by Polarized Egress

Spread of Hepatitis B Viruses In Vitro Requires Extracellular Progeny and May Be Codetermined by Polarized Egress

Acute Liver Injury Following Infection With A Cytopathic Strain of Duck Hepatitis B Virus

Frequency of Spontaneous Mutations in an Avian Hepadnavirus Infection

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