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Hepatitis D virus replication is sensed by MDA5 and induces IFN-β/λ responses in hepatocytes

  • Zhenfeng Zhang
    Affiliations
    Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
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  • Christina Filzmayer
    Affiliations
    Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
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  • Yi Ni
    Affiliations
    Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
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  • Holger Sültmann
    Affiliations
    Cancer Genome Research Group, German Cancer Research Center (DKFZ) and National Center for Tumor Diseases (NCT), Im Neuenheimer Feld 460, Heidelberg, Germany

    Translational Lung Research Center (TLRC), German Center for Lung Research (DZL), Heidelberg, Germany

    German Cancer Consortium (DKTK), Heidelberg, Germany
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  • Pascal Mutz
    Affiliations
    Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany

    Division of Virus-Associated Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg, Germany
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  • Marie-Sophie Hiet
    Affiliations
    Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
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  • Florian W.R. Vondran
    Affiliations
    Regenerative Medicine and Experimental Surgery (ReMediES), Department of General, Visceral and Transplant Surgery, Hannover Medical School, Hannover, Germany

    German Center for Infection Research (DZIF), Partner Site Hannover-Braunschweig, Hannover, Germany
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  • Ralf Bartenschlager
    Affiliations
    Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany

    German Center for Infection Research (DZIF) - Heidelberg Partner Site, Heidelberg, Germany

    Division of Virus-Associated Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg, Germany
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  • Stephan Urban
    Correspondence
    Corresponding author. Address: Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Im Neuenheimer Feld 344, D-69120 Heidelberg, Germany. Tel.: +49 6221 564902; fax: +49 6221 561946.
    Affiliations
    Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany

    German Center for Infection Research (DZIF) - Heidelberg Partner Site, Heidelberg, Germany
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Published:March 07, 2018DOI:https://doi.org/10.1016/j.jhep.2018.02.021

      Highlights

      • In contrast to HBV, HDV activates the IFN response in hepatocytes.
      • MDA5 is the key pattern recognition receptor sensing HDV replication.
      • HDV replication is insensitive to the MDA5-mediated self-induced IFN response.
      • IFN treatment doesn’t abolish intracellular HDV replication in vitro.

      Background & Aims

      Hepatitis B virus (HBV) and D virus (HDV) co-infections cause the most severe form of viral hepatitis. HDV induces an innate immune response, but it is unknown how the host cell senses HDV and if this defense affects HDV replication. We aim to characterize interferon (IFN) activation by HDV, identify the responsible sensor and evaluate the effect of IFN on HDV replication.

      Methods

      HDV and HBV susceptible hepatoma cell lines and primary human hepatocytes (PHH) were used for infection studies. Viral markers and cellular gene expression were analyzed at different time points after infection. Pattern recognition receptors (PRRs) required for HDV-mediated IFN activation and the impact on HDV replication were studied using stable knock-down or overexpression of the PRRs.

      Results

      Microarray analysis revealed that HDV but not HBV infection activated a broad range of interferon stimulated genes (ISGs) in HepG2NTCP cells. HDV strongly activated IFN-β and IFN-λ in cell lines and PHH. HDV induced IFN levels remained unaltered upon RIG-I (DDX58) or TLR3 knock-down, but were almost completely abolished upon MDA5 (IFIH1) depletion. Conversely, overexpression of MDA5 but not RIG-I and TLR3 in HuH7.5NTCP cells partially restored ISG induction. During long-term infection, IFN levels gradually diminished in both HepG2NTCP and HepaRGNTCP cell lines. MDA5 depletion had little effect on HDV replication despite dampening HDV-induced IFN response. Moreover, treatment with type I or type III IFNs did not abolish HDV replication.

      Conclusion

      Active replication of HDV induces an IFN-β/λ response, which is predominantly mediated by MDA5. This IFN response and exogenous IFN treatment have only a moderate effect on HDV replication in vitro indicating the adaption of HDV replication to an IFN-activated state.

      Lay summary

      In contrast to hepatitis B virus, infection with hepatitis D virus induces a strong IFN-β/λ response in innate immune competent cell lines. MDA5 is the key sensor for the recognition of hepatitis D virus replicative intermediates. An IFN-activated state did not prevent hepatitis D virus replication in vitro, indicating that hepatitis D virus is resistant to self-induced innate immune responses and therapeutic IFN treatment.

      Graphical abstract

      Keywords

      Linked Article

      • New insights into HDV-induced innate immunity: MDA5 senses HDV replication
        Journal of HepatologyVol. 69Issue 1
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          Coinfection with hepatitis D virus (HDV) is observed in about 5–10% of hepatitis B virus (HBV)-infected individuals. It is known to be associated with more severe and faster progressing liver disease, as well as poor prognosis.1 Under certain conditions a strong innate immune response to a viral infection can lead to severe immunopathogenesis.2 For HDV, neither the factors that drive the pathogenesis in HDV/HBV coinfection, nor the details of the interaction between HDV and the host innate immune system have been well characterized.
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