Antigen-specific blastogenesis assays for duck hepatitis B virus using duck peripheral blood and splenic mononuclear cells

ABSTRACT Residual hepatitis B virus (HBV) DNA can be detected in serum and liver after apparent recovery from transient infection. However, it is not known if this residual HBV DNA represents ongoing viral replication and antigen expression. In the current study, ducks inoculated with duck hepatitis B virus (DHBV) were monitored for residual DHBV DNA following recovery from transient infection until 9 months postinoculation (p.i.). Resolution of DHBV infection occurred in 13 out of 15 ducks by 1-month p.i., defined as clearance of DHBV surface antigen-positive hepatocytes from the liver and development of anti-DHBV surface antibodies. At 9 months p.i., residual DHBV DNA was detected using nested PCR in 10/11 liver, 7/11 spleen, 2/11 kidney, 1/11 heart, and 1/11 adrenal samples. Residual DHBV DNA was not detected in serum or peripheral blood mononuclear cells. Within the liver, levels of residual DHBV DNA were 0.0024 to 0.016 copies per cell, 40 to 80% of which were identified as covalently closed circular viral DNA by quantitative PCR assay. This result, which was confirmed by Southern blot hybridization, is consistent with suppressed viral replication or inactive infection. Samples of liver and spleen cells from recovered animals did not transmit DHBV infection when inoculated into 1- to 2-day-old ducklings, and immunosuppressive treatment of ducks with cyclosporine and dexamethasone for 4 weeks did not alter levels of residual DHBV DNA in the liver. These findings further characterize a second form of hepadnavirus persistence in a suppressed or inactive state, quite distinct from the classical chronic carrier state.

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Faculty Opinions recommendation of Translation of duck hepatitis B virus reverse transcriptase by ribosomal shunting.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1021978.247821 ◽

2004 ◽

Author(s):

Don Coen

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Reverse Transcriptase ◽

Duck Hepatitis B Virus ◽

Duck Hepatitis ◽

B Virus

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Integrated structures of duck hepatitis B virus DNA in hepatocellular carcinoma.

Journal of Virology ◽

10.1128/jvi.62.3.861-865.1988 ◽

1988 ◽

Vol 62 (3) ◽

pp. 861-865 ◽

Cited By ~ 13

Author(s):

F Imazeki ◽

K Yaginuma ◽

M Omata ◽

K Okuda ◽

M Kobayashi ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Hepatitis B Virus ◽

Hepatitis B ◽

Duck Hepatitis B Virus ◽

Hepatitis B Virus Dna ◽

Duck Hepatitis ◽

B Virus ◽

Integrated Structures

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Different forms of hepatitis B virus DNA and expression of HBV antigens in peripheral blood mononuclear cells in chronic hepatitis B

Journal of Medical Virology ◽

10.1002/jmv.1890310413 ◽

1990 ◽

Vol 31 (4) ◽

pp. 312-317 ◽

Cited By ~ 25

Author(s):

P. Bouffard ◽

J.-P. Lamelin ◽

F. Zoulim ◽

C. Pichoud ◽

C. Trepo

Keyword(s):

Chronic Hepatitis ◽

Hepatitis B Virus ◽

Hepatitis B ◽

Chronic Hepatitis B ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

B Virus ◽

Blood Mononuclear Cells

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Fine Mapping of Neutralization Epitopes on Duck Hepatitis B Virus (DHBV) pre-S Protein Using Monoclonal Antibodies and Overlapping Peptides

Virology ◽

10.1006/viro.1993.1024 ◽

1993 ◽

Vol 192 (1) ◽

pp. 217-223 ◽

Cited By ~ 23

Author(s):

Sylvie Chassot ◽

Véronique Lambert ◽

Alan Kay ◽

Catherine Godinot ◽

Bernard Roux ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Hepatitis B Virus ◽

Hepatitis B ◽

Fine Mapping ◽

Duck Hepatitis B Virus ◽

S Protein ◽

Duck Hepatitis ◽

B Virus

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Duck hepatitis B virus: Cloning and subcloning of the viral genome

Zentralblatt für Bakteriologie Mikrobiologie und Hygiene Series A Medical Microbiology Infectious Diseases Virology Parasitology ◽

10.1016/s0176-6724(89)80012-4 ◽

1989 ◽

Vol 270 (3) ◽

pp. 424-433

Author(s):

Konrad Oexle ◽

Hubert E. Blum ◽

Eike Walter ◽

Wolf-Bernhard Offensperger ◽

Silke Offensperger ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Viral Genome ◽

Duck Hepatitis B Virus ◽

Duck Hepatitis ◽

B Virus

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Duck hepatitis B virus: a model to assess efficacy of disinfectants against hepadnavirus infectivity

Epidemiology and Infection ◽

10.1017/s0950268800067480 ◽

1991 ◽

Vol 106 (3) ◽

pp. 435-443 ◽

Cited By ~ 33

Author(s):

S. M. Murray ◽

J. S. Freiman ◽

K. Vickery ◽

D. Lim ◽

Y. E. Cossart ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Room Temperature ◽

Virus Titre ◽

Experimental Protocol ◽

Duck Hepatitis B Virus ◽

Duck Hepatitis ◽

B Virus ◽

Undiluted Serum ◽

Sterile Filter

SUMMARYThe efficacy of three proprietary glutaraldehyde disinfectants and their component bases was assessed using the duck hepatitis B virus (DHBV) model. Inactivation of infectivity of undiluted serum containing 106·8ID50/ml DHBV was assessed after a mixture with an equal volume of disinfectant had stood at room temperature for 10 min. A dried spill of infectious serum was simulated using sterile filter paper disks, saturated with serum containing DHBV, dried and then exposed to test disinfectant for 10 min. Residual infectivity, and hence the reduction in virus titre, was determined by inoculation of dilutions of the treated samples into 1-day-old ducklings. A greater than 3 log10reduction in virus titre could be demonstrated for the disinfectants as well as for some of their component bases. Disinfectant activity varied according to the method of viral presentation but a reduction of exposure time from 10 to 2·5 min did not diminish activity. The experimental protocol permits a comparative and quantitative assessment of the efficacy of both established and new disinfectants.

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Rise in gamma interferon expression during resolution of duck hepatitis B virus infection

Journal of General Virology ◽

10.1099/vir.0.82170-0 ◽

2006 ◽

Vol 87 (11) ◽

pp. 3225-3232 ◽

Cited By ~ 14

Author(s):

Ramamurthy Narayan ◽

Thierry Buronfosse ◽

Ursula Schultz ◽

Philippe Chevallier-Gueyron ◽

Sylviane Guerret ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Mononuclear Cells ◽

Reference Model ◽

Gamma Interferon ◽

Strong Increase ◽

Moderate Increase ◽

Histological Changes ◽

Duck Hepatitis ◽

B Virus

Gamma interferon (IFN-γ) expression plays a crucial role in the control of mammalian hepatitis B virus (HBV) infection. However, the role of duck INF-γ (DuIFN-γ) in the outcome of duck HBV (DHBV) infection, a reference model for hepadnavirus replication studies, has not yet been investigated. This work explored the dynamics of DuIFN-γ expression in liver and peripheral blood mononuclear cells (PBMCs) during resolution of DHBV infection in adolescent ducks in relation to serum and liver markers of virus replication, histological changes and humoral response induction. DHBV infection of 3-week-old ducks resulted in transient expression of intrahepatic preS protein (days 3–14) and mild histological changes. Low-level viraemia was detected only during the first 10 days of infection and was accompanied by early anti-preS antibody response induction. Importantly, a strong increase in intrahepatic DuIFN-γ RNA was detected by real-time RT-PCR at days 6–14, which coincided with a sharp decrease in both viral DNA and preS protein in the liver. Interestingly, liver DuIFN-γ expression remained augmented to the end of the follow-up period (day 66) and correlated with portal lymphocyte infiltration and persistence of trace quantities of intrahepatic DHBV DNA in animals that had apparently completely resolved the infection. Moreover, in infected ducks, a moderate increase was detected in the levels of DuIFN-γ in PBMCs (days 12–14), which coincided with the peak in liver DuIFN-γ RNA levels. These data reveal that increased DuIFN-γ expression in liver and PBMCs is concomitant with viral clearance, characterizing the resolution of infection, and provide new insights into the host–virus interactions that control DHBV infection.

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