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Journal of Virology, October 2005, p. 12242-12252, Vol. 79, No. 19
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.19.12242-12252.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Covalently Closed Circular DNA Is the Predominant Form of Duck Hepatitis B Virus DNA That Persists following Transient Infection

Marc F. Le Mire,1,{dagger} Darren S. Miller,1 Wendy K. Foster,1 Christopher J. Burrell,1,2 and Allison R. Jilbert1,2*

School of Molecular and Biomedical Science, University of Adelaide, North Terrace, Adelaide, South Australia 5005, Australia,1 Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Frome Road, Adelaide, South Australia 5000, Australia2

Received 17 March 2005/ Accepted 5 July 2005

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.

* Corresponding author. Mailing address: Hepatitis Virus Research Laboratory, Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Frome Rd. (P.O. Box 14 Rundle Mall), Adelaide, South Australia 5000, Australia. Phone: 61 8 8303 5399. Fax: 61 8 8303 7532. E-mail: allison.jilbert{at}adelaide.edu.au.

{dagger} Present address: Department of Gastroenterology, Hepatology and General Medicine, Royal Adelaide Hospital, North Terrace, Adelaide, South Australia 5005, Australia.

Journal of Virology, October 2005, p. 12242-12252, Vol. 79, No. 19
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.19.12242-12252.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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