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Role of core protein mutations in the development of occult HBV infection

  • Author Footnotes
    † These authors contributed equally to the work.
    Jingna Chen
    Footnotes
    † These authors contributed equally to the work.
    Affiliations
    Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China

    Department of Laboratory Medicine, General Hospital of Southern Theatre Command of PLA, Guangzhou, China

    Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Shantou, China
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  • Author Footnotes
    † These authors contributed equally to the work.
    Bochao Liu
    Footnotes
    † These authors contributed equally to the work.
    Affiliations
    Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China
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  • Xi Tang
    Affiliations
    Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China

    Department of Infectious Diseases, The First Foshan People’s Hospital, Foshan, China
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  • Xin Zheng
    Affiliations
    Shenzhen Blood Center, Shenzhen, China
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  • Jinhui Lu
    Affiliations
    Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China
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  • Ling Zhang
    Affiliations
    Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China
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  • Wenjing Wang
    Affiliations
    Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China
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  • Daniel Candotti
    Affiliations
    Department of Blood Transmitted Agents, National Institute of Blood Transfusion, Paris, France
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  • Yongshui Fu
    Affiliations
    Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China

    Guangzhou Blood Center, Guangzhou, China
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  • Jean-Pierre Allain
    Affiliations
    Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China

    Department of Haematology, University of Cambridge, Cambridge, UK
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  • Chengyao Li
    Correspondence
    Corresponding authors. Addresses: Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou 510515, China; Tel./Fax: +86-20-61648466
    Affiliations
    Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China
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  • Linhai Li
    Correspondence
    Department of Laboratory Medicine, PLA General Hospital of Southern Theatre Command, Guangzhou 510010, China
    Affiliations
    Department of Laboratory Medicine, General Hospital of Southern Theatre Command of PLA, Guangzhou, China
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  • Tingting Li
    Correspondence
    Corresponding authors. Addresses: Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou 510515, China; Tel./Fax: +86-20-61648466
    Affiliations
    Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally to the work.
Published:January 13, 2021DOI:https://doi.org/10.1016/j.jhep.2020.12.023

      Highlights

      • Amino acid (aa) substitutions in the core protein (Cp) of occult HBV infection (OBI) strains were identified.
      • An OBI mutant strain (SZA) carrying 9 aa substitutions in Cp was characterised in vitro (cells) and in vivo (mice).
      • The functional impact of individual aa substitutions in pHBV1.3-SZA Cp replicons was assessed in vitro and in vivo.
      • The Cp W62R mutation significantly reduced HBV protein production.
      • The Cp W62R and its combination mutations might contribute to the occurrence of OBI.

      Background & Aims

      Occult HBV infection (OBI) is associated with transfusion-transmitted HBV infection and hepatocellular carcinoma. Studies on OBI genesis have concentrated on mutations in the S region and the regulatory elements. Herein, we aimed to determine the role of mutations in the core region on OBIs.

      Methods

      An OBI strain (SZA) carrying 9 amino acid (aa) substitutions in the core protein/capsid (Cp) was selected by sequence alignment and Western blot analysis from 26 genotype B OBI samples to extensively explore the impact of Cp mutations on viral antigen production in vitro and in vivo.

      Results

      A large panel of 30 Cp replicons were generated by a replication-competent pHBV1.3 carrying SZA or wild-type (WT) Cp in a 1.3-fold over-length of HBV genome, in which the various Cp mutants were individually introduced by repairing site mutations of SZA-Cp or creating site mutations of WT-Cp by site-directed mutagenesis. The expression of HBcAg, HBeAg, and HBsAg and viral RNA was quantified from individual SZA and WT Cp mutant replicons in transfected Huh7 cells or infected mice, respectively. An analysis of the effect of Cp mutants on intracellular or extracellular viral protein production indicated that the W62R mutation in Cp had a critical impact on the reduction of HBcAg and HBeAg production during HBV replication, whereas P50H and/or S74G mutations played a limited role in influencing viral protein production in vivo.

      Conclusions

      W62R and its combination mutations in HBV Cp might massively affect HBcAg and HBeAg production during viral replication, which, in turn, might contribute to the occurrence of OBI.

      Lay summary

      Occult hepatitis B virus infections (OBIs) have been found to be associated with amino acid mutations in the S region of the HBV, but the role of mutations in the core protein (Cp) remains unclear. In this study, an OBI strain (SZA) carrying 9 amino acid substitutions in Cp has been examined comprehensively in vitro and in vivo. The W62R mutation in Cp majorly reduces HBcAg and HBeAg production during HBV replication, potentially contributing to the occurrence of OBI.

      Graphical abstract

      Keywords

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