Interferon prevents formation of replication-competent hepatitis B virus RNA-containing nucleocapsids.

We have previously shown that IFN-beta inhibits hepatitis B virus (HBV) replication by noncytolytic mechanisms that either destabilize pregenomic (pg)RNA-containing capsids or prevent their assembly. Using immortalized murine hepatocyte cell lines stably transfected with a doxycycline (dox)-inducible HBV replication system, we now show that replication-competent pgRNA-containing capsids are not produced when the cells are pretreated with IFN-beta before HBV expression is induced with dox. Furthermore, the turnover rate of preformed HBV RNA-containing capsids is not changed in the presence of IFN-beta or IFN-gamma under conditions in which further pgRNA synthesis is inhibited by dox removal. In summary, these results demonstrate that types 1 and 2 IFN activate hepatocellular mechanism(s) that prevent the formation of replication-competent HBV capsids and, thereby, inhibit HBV replication.

[1]  F. Zoulim,et al.  Reverse transcription in hepatitis B viruses is primed by a tyrosine residue of the polymerase , 1994, Journal of virology.

[2]  F. Chisari,et al.  Intracellular inactivation of the hepatitis B virus by cytotoxic T lymphocytes. , 1996, Immunity.

[3]  D. Trono,et al.  Inhibition of Hepatitis B Virus Replication by APOBEC3G , 2004, Science.

[4]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[5]  F. Spagnoli,et al.  Identification of a Bipotential Precursor Cell in Hepatic Cell Lines Derived from Transgenic Mice Expressing Cyto-Met in the Liver , 1998, The Journal of cell biology.

[6]  V. Pasquetto,et al.  Cytokine-Sensitive Replication of Hepatitis B Virus in Immortalized Mouse Hepatocyte Cultures , 2002, Journal of Virology.

[7]  M. Shapiro,et al.  Viral clearance without destruction of infected cells during acute HBV infection. , 1999, Science.

[8]  A. Vitiello,et al.  HLA A2 restricted cytotoxic T lymphocyte responses to multiple hepatitis B surface antigen epitopes during hepatitis B virus infection. , 1993, Journal of immunology.

[9]  F. Chisari,et al.  HLA-A31- and HLA-Aw68-restricted cytotoxic T cell responses to a single hepatitis B virus nucleocapsid epitope during acute viral hepatitis , 1993, The Journal of experimental medicine.

[10]  F. Chisari,et al.  HLA class I-restricted human cytotoxic T cells recognize endogenously synthesized hepatitis B virus nucleocapsid antigen. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[11]  F. Chisari,et al.  Expression of hepatitis B virus surface and core antigens: influences of pre-S and precore sequences , 1987, Journal of virology.

[12]  J. Li,et al.  Roles of the three major phosphorylation sites of hepatitis B virus core protein in viral replication. , 1999, Virology.

[13]  F. Chisari,et al.  Identification of immunodominant T cell epitopes of the hepatitis B virus nucleocapsid antigen. , 1991, The Journal of clinical investigation.

[14]  Response to Comment on "Observation of the Inverse Doppler Effect" , 2004, Science.

[15]  M. Nassal,et al.  Reconstitution of a Functional Duck Hepatitis B Virus Replication Initiation Complex from Separate Reverse Transcriptase Domains Expressed in Escherichia coli , 2001, Journal of Virology.

[16]  J. Wands,et al.  Hepatitis B virus mutants associated with 3TC and famciclovir administration are replication defective , 1998, Hepatology.

[17]  E. Gazina,et al.  Core Protein Phosphorylation Modulates Pregenomic RNA Encapsidation to Different Extents in Human and Duck Hepatitis B Viruses , 2000, Journal of virology.

[18]  C. Seeger,et al.  Hsp90 is required for the activity of a hepatitis B virus reverse transcriptase. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[19]  M. Biermer,et al.  Tumor Necrosis Factor Alpha Inhibition of Hepatitis B Virus Replication Involves Disruption of Capsid Integrity through Activation of NF-κB , 2003, Journal of Virology.

[20]  R. Purcell,et al.  Expansion and contraction of the hepatitis B virus transcriptional template in infected chimpanzees , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[21]  R. Bartenschlager,et al.  Expression of the P-protein of the human hepatitis B virus in a vaccinia virus system and detection of the nucleocapsid-associated P-gene product by radiolabelling at newly introduced phosphorylation sites. , 1992, Nucleic acids research.

[22]  C. Seeger,et al.  Hepadnavirus assembly and reverse transcription require a multi‐component chaperone complex which is incorporated into nucleocapsids , 1997, The EMBO journal.

[23]  F. Chisari,et al.  High-level hepatitis B virus replication in transgenic mice , 1995, Journal of virology.

[24]  F. Chisari,et al.  Hepatitis B virus immunopathogenesis. , 1995, Annual review of immunology.

[25]  M. Malim,et al.  Comment on "Inhibition of Hepatitis B Virus Replication by APOBEC3G" , 2004, Science.

[26]  R. Bartenschlager,et al.  The P gene product of hepatitis B virus is required as a structural component for genomic RNA encapsidation , 1990, Journal of virology.

[27]  F. Chisari,et al.  Intrahepatic Induction of Alpha/Beta Interferon Eliminates Viral RNA-Containing Capsids in Hepatitis B Virus Transgenic Mice , 2000, Journal of Virology.

[28]  L. Wodicka,et al.  Genomic analysis of the host response to hepatitis C virus infection , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[29]  M. Gossen,et al.  Transcriptional activation by tetracyclines in mammalian cells. , 1995, Science.

[30]  H. Blum,et al.  Hepatitis B Virus Nucleocapsids Formed by Carboxy-Terminally Mutated Core Proteins Contain Spliced Viral Genomes but Lack Full-Size DNA , 2004, Journal of Virology.

[31]  R. Purcell,et al.  CD8+ T Cells Mediate Viral Clearance and Disease Pathogenesis during Acute Hepatitis B Virus Infection , 2003, Journal of Virology.

[32]  F. Chisari,et al.  Relative Sensitivity of Hepatitis B Virus and Other Hepatotropic Viruses to the Antiviral Effects of Cytokines , 2000, Journal of Virology.

[33]  R. König,et al.  Species-Specific Exclusion of APOBEC3G from HIV-1 Virions by Vif , 2003, Cell.

[34]  F. Chisari,et al.  Signal transduction pathways that inhibit hepatitis B virus replication. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[35]  M. Imamura,et al.  G to A hypermutation of hepatitis B virus , 2005, Hepatology.

[36]  B. Moss,et al.  The cytotoxic T lymphocyte response to multiple hepatitis B virus polymerase epitopes during and after acute viral hepatitis , 1995, The Journal of experimental medicine.

[37]  J. Pollack,et al.  Site-specific RNA binding by a hepatitis B virus reverse transcriptase initiates two distinct reactions: RNA packaging and DNA synthesis , 1994, Journal of virology.

[38]  J. Pollack,et al.  An RNA stem-loop structure directs hepatitis B virus genomic RNA encapsidation , 1993, Journal of virology.

[39]  J. Tavis,et al.  Hepadnavirus reverse transcription initiates within the stem-loop of the RNA packaging signal and employs a novel strand transfer , 1994, Journal of virology.