Duck Hepatitis B Virus Replication in Primary Bile Duct Epithelial Cells

Lee, Jia-Yee; Culvenor, Janetta G.; Angus, Peter W; Smallwood, Richard A.; Nicoll, Amanda; Locarnini, Stephen

doi:10.1128/jvi.75.16.7651-7661.2001

Cited by 16 publications

(14 citation statements)

References 46 publications

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“…On day 12, the cells were fixed with absolute methanol for 5 min at room temperature (RT). The fixed cells were processed for single-labeled or dual-labeled immunofluorescence assay (IFA) as described previously (13). For dual-labeled IFA, the cells were incubated for 30 min at 37°C with or without mouse monoclonal antibody to HBV pre-S2 antigen (Abcam Ltd., Cambridge, United Kingdom) diluted 1/100 in PBS.…”

Section: Methodsmentioning

confidence: 99%

“…Coverslip preparations were mounted in fluorescent mounting medium (Dako Corp.) and viewed with a Bio-Rad (Hemel Hempstead, United Kingdom) MRC 1024 laser confocal system attached to a Zeiss microscope. Image collection parameters were adjusted to minimize cross-channel leak over and tested using appropriate single-and dual-labeled preparations (13).…”

Section: Methodsmentioning

confidence: 99%

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Endocytosis of Hepatitis B Immune Globulin into Hepatocytes Inhibits the Secretion of Hepatitis B Virus Surface Antigen and Virions

Schilling¹,

Ijaz

Davidoff

et al. 2003

J Virol

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Hepatitis B immunoglobulin is used for prophylaxis against hepatitis B virus (HBV) and is thought to act by neutralization of virions and hepatitis B virus surface antigen (HBsAgHepatitis B immunoglobulin (HBIG) is used clinically as passive immunoprophylaxis for accidental exposure to hepatitis B virus (HBV) and long term to prevent HBV recurrence in the graft after liver transplantation (24). It contains high-titer antibodies against HBV surface antigen (HBsAg), which is the major component of the outer envelope of the 42-nm-diameter hepatitis B virion, as well as the 22-nm-diameter subviral particles. The therapeutic effect of HBIG is believed to be due to high-affinity binding with HBs-containing particles and neutralization of HBV in the circulation.Despite HBIG prophylaxis, HBV infection recurs in 30% of patients who receive transplants for HBsAg-positive cirrhosis (24). The failure of immunoprophylaxis is due either to a high HBV load and inadequate neutralization by HBIG or to the emergence of antibody-induced escape HBV mutants (3, 6, 25). These mutant HBV strains contain amino acid substitutions within the conserved a-determinant (a group-reactive region between amino acids 124 and 149 of HBsAg), which abrogate the binding affinity of anti-HBs (4, 5, 22, 27). The most frequent mutation occurs at codon 145 of the surface open reading frame, leading to a glycine (G)-to-arginine (R) substitution-G145R-which has been shown to emerge both in liver transplant patients receiving HBIG prophylaxis and in HBV vaccine recipients (2,6,22). The mechanism for the emergence of HBsAg mutations that escape antibody recognition has not been defined.Earlier studies have demonstrated the presence of membrane-bound and/or nuclear localization of immunoglobulin G (IgG) in hepatocytes of patients with chronic HBV infection, who express HBV core antigen or hepatitis delta virus antigen in the liver (17,19,23). Recently, a novel Fc receptor for IgG (FcRn) which mediates the transcytosis of IgG from serum to bile and protects the internalized IgG from catabolism has been identified on the plasma membranes of adult rat hepatocytes (1, 9). FcRn is a heterodimer of ␤ 2 -microglobulin light chain and a major histocompatibility complex class I-like heavy chain that binds IgG via Fc residues in a pH-dependent manner. IgG binding to FcRn is followed by endocytosis of the complex in the acidic endosome environment, trafficking through cellular conduits to bypass lysosomal activities and finally releasing IgG in the extracellular fluids (7). Whether hepatitis B immunoglobulin enters HBV-infected hepatocytes and whether an interaction with HBsAg occurs within cells, in addition to the interaction in serum, have not been investigated.In the present study, we investigated the hypothesis that HBIG is able to bind to hepatocytes and affect the secretion of HBsAg and HBV virions from the cells. For this purpose, we used a panel of human hepatocyte-derived cell lines cultured together with monoclonal and polyclonal HBs-specific antibodies. The...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Endocytosis of Hepatitis B Immune Globulin into Hepatocytes Inhibits the Secretion of Hepatitis B Virus Surface Antigen and Virions

Schilling¹,

Ijaz

Davidoff

et al. 2003

J Virol

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“…Since elevated levels of cccDNA (Ͼ50 copies per infected cell) have previously been found only in cells infected with cytopathic strains of DHBV (19) and not in wild-type DHBV infection (13,14), this result suggests either that cccDNA is present in hepatocytes with undetectable levels of DHBsAg, i.e., cccDNA is transcriptionally inactive, or that cccDNA is present at higher copy numbers in nonhepatocyte cell types (e.g., bile duct cells). Bile duct cells have been reported to contain increased levels of DHBsAg and DHBV DNA (14,18,26), but quantitation of cccDNA copy number per infected bile duct cell in vivo has not been performed.…”

Section: Vol 47 2003mentioning

confidence: 99%

Entecavir Therapy Combined with DNA Vaccination for Persistent Duck Hepatitis B Virus Infection

Foster

Miller

Marion

et al. 2003

Antimicrob Agents Chemother

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This study was designed to test the efficacy of antiviral treatment with entecavir (ETV) in combination with DNA vaccines expressing duck hepatitis B virus (DHBV) antigens as a therapy for persistent DHBV infection in ducks. Ducks were inoculated with 109 DHBV genomes at 7 days of age, leading to widespread infection of the liver and viremia within 7 days, and were then treated orally with either ETV (0.1 mg/kg of body weight/day) or distilled water from 21 days posthatch for 244 days. Treatment with ETV caused a 4-log drop in serum DHBV DNA levels within 80 days and a slower 2-to 3-log drop in serum DHBV surface antigen (DHBsAg) levels within 120 days. Following withdrawal of ETV, levels of serum DHBV DNA and DHBsAg rebounded to match those in the water-treated animals within 40 days. Sequential liver biopsy samples collected throughout the study showed that ETV treatment reduced DHBV DNA replicative intermediates 70-fold in the liver, while the level of the stable, template form, covalently closed circular DNA decreased only 4-fold. ETV treatment reduced both the intensity of antigen staining and the percentage of antigen-positive hepatocytes in the liver, but the intensity of antigen staining in bile duct cells appeared not to be effected. Intramuscular administration of five doses of a DNA vaccine expressing the DHBV presurface, surface, precore, and core antigens, both alone and concurrently with ETV treatment, on days 50, 64, 78, 127, and 141 did not result in any significant effect on viral markers.Hepatitis B virus (HBV) is a noncytopathic virus that replicates primarily in hepatocytes. HBV infection of humans can result in either a transient infection with development of neutralizing antibodies and immunity to reinfection or a persistence of viral infection for many years. Patients with either acute or persistent HBV infection often have extensive infection of the liver with high titers of infectious virus and noninfectious surface antigen particles circulating in the bloodstream. Immune responses to HBV are responsible for clearance of virus-infected cells from the liver and for the liver damage seen in both acute and persistent infection. Models of hepadnavirus pathogenesis usually propose that effective humoral and cell-mediated immune responses lead to recovery from infection with elimination of virally infected cells or replicative intermediates, while ineffective immune responses allow extensive viral replication with or without hepatocyte damage (reviewed in references 2 and 3). Thus, in individuals destined to become persistently infected, the high viral load may overwhelm the capacity of the immune system or favor the induction of ineffective antibody responses.Persistent HBV infection occurs in 5 to 10% of adults with acute infections and in Ͼ90% of neonates. Current approaches to treatment for persistent HBV infection include long-term alpha interferon administration and antiviral drug therapy aimed at inhibiting viral replication. Partial suppression of levels of circulating virus can be ach...

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“…7). From that, we can conclude that speciesspecific factors necessary for certain steps of DHBV infection following penetration are present not only in hepatocytes but also in kidney cells and, as described by others, in primary bile duct epithelial cells (42), in the yolk sacs of embryos (61), or in the pancreases of duck embryos and ducklings (27,42,61).…”

Section: Discussionmentioning

confidence: 96%

Entry of Duck Hepatitis B Virus into Primary Duck Liver and Kidney Cells after Discovery of a Fusogenic Region within the Large Surface Protein

Maenz

Chang

Iwanski

et al. 2007

J Virol

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Hepatitis B viruses exhibit a narrow host range specificity that is believed to be mediated by a domain of the large surface protein, designated L. For duck hepatitis B virus, it has been shown that the pre-S domain of L binds to carboxypeptidase D, a cellular receptor present in many species on a wide variety of cell types. Nonetheless, only hepatocytes become infected. It has remained vague which viral features determine host range specificity and organotropicity. By using chymotrypsin to treat duck hepatitis B virus, we addressed the question of whether a putative fusogenic region within the amino-terminal end of the small surface protein may participate in viral entry and possibly constitute one of the determinants of the host range of the virus. Addition of the enzyme to virions resulted in increased infectivity. Remarkably, even remnants of enzyme-treated subviral particles proved to be inhibitory to infection. A noninfectious deletion mutant devoid of the binding region for carboxypeptidase D could be rendered infectious for primary duck hepatocytes by treatment with chymotrypsin. Although because of the protease treatment mutant and wild-type viruses may have become infectious in an unspecific and receptor-independent manner, their host range specificity was not affected, as shown by the inability of the virus to replicate in different hepatoma cell lines, as well as primary chicken hepatocytes. Instead, the organotropicity of the virus could be reduced, which was demonstrated by infection of primary duck kidney cells.The family Hepadnaviridae, with the human hepatitis B virus (HBV) as the prototype member, consists of small, DNAcontaining, enveloped viruses which cause liver diseases in humans, rodents, and birds (22). A characteristic feature of HBV and other hepadnaviruses is a strong species specificity where, e.g., HBV infects humans and chimpanzees (1) and duck HBV (DHBV) infects only certain duck and goose species (49). Moreover, all of the hepadnaviruses show relatively strict organotropicity, infecting mainly hepatocytes.Facilitated by the identification of neutralizing epitopes (12) and the allocation of host range-specific determinants to the pre-S region of L (29), it was possible to demonstrate the binding of DHBV to a putative hepatocellular receptor molecule via a certain domain on pre-S (7,8,30,39,63). This receptor molecule has been identified as gp180, a transmembrane protein of the carboxypeptidase D family that, however, has also been found in nonliver duck tissues that are not susceptible to DHBV infection (16,38,62). Furthermore, it has been shown that LMH cells, a chicken hepatoma cell line which resists infection but is capable of virus production after transfection of full-length DHBV DNA, also express gp180 (38). In addition, it was demonstrated that this molecule is concentrated in the Golgi apparatus, to cycle to and from the plasma membrane, where it colocalizes with pre-S (6, 7). Expression of duck gp180 in HuH7 cells, a human hepatoma cell line, led merely to internalizatio...

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Duck Hepatitis B Virus Replication in Primary Bile Duct Epithelial Cells

Cited by 16 publications

References 46 publications

Endocytosis of Hepatitis B Immune Globulin into Hepatocytes Inhibits the Secretion of Hepatitis B Virus Surface Antigen and Virions

Endocytosis of Hepatitis B Immune Globulin into Hepatocytes Inhibits the Secretion of Hepatitis B Virus Surface Antigen and Virions

Entecavir Therapy Combined with DNA Vaccination for Persistent Duck Hepatitis B Virus Infection

Entry of Duck Hepatitis B Virus into Primary Duck Liver and Kidney Cells after Discovery of a Fusogenic Region within the Large Surface Protein

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