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Multimodal investigation of rat hepatitis E virus antigenicity: Implications for infection, diagnostics, and vaccine efficacy

  • Siddharth Sridhar
    Correspondence
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Queen Mary Hospital, Pokfulam, Hong Kong. Tel.: (852) 22552396.
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China

    State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong, Hong Kong

    Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong
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  • Author Footnotes
    † Contributed equally.
    Jianwen Situ
    Footnotes
    † Contributed equally.
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
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  • Author Footnotes
    † Contributed equally.
    Jian-Piao Cai
    Footnotes
    † Contributed equally.
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
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  • Author Footnotes
    † Contributed equally.
    Cyril Chik-Yan Yip
    Footnotes
    † Contributed equally.
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
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  • Shusheng Wu
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
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  • Anna Jin-Xia Zhang
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
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  • Lei Wen
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
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  • Nicholas Foo-Siong Chew
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
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  • Wan-Mui Chan
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
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  • Rosana Wing-Shan Poon
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China
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  • Jasper Fuk-Woo Chan
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China

    State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong, Hong Kong

    Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong
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  • Dominic Ngai-Chong Tsang
    Affiliations
    Public Health Laboratory Services Branch, Department of Health, Hong Kong
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  • Honglin Chen
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China

    State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong, Hong Kong

    Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong
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  • Ning-Shao Xia
    Affiliations
    School of Life Sciences, Xiamen University, Xiamen, China
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  • Kwok-Yung Yuen
    Correspondence
    Corresponding authors. Addresses: Carol Yu Centre for Infection, State Key Laboratory of Emerging Infectious Diseases, Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Queen Mary Hospital, Pokfulam, Hong Kong. Tel.: (852) 22554892. Fax: (852) 28551241.
    Affiliations
    Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China

    State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong, Hong Kong

    Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong

    The Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The University of Hong Kong, Hong Kong
    Search for articles by this author
  • Author Footnotes
    † Contributed equally.
Published:April 09, 2021DOI:https://doi.org/10.1016/j.jhep.2020.12.028

      Highlights

      • Rat HEV (HEV-C1) is antigenically distinct from human HEV genotypes.
      • Human HEV-based antigen and antibody assays may not diagnose HEV-C1 infection.
      • Prior exposure to human HEV genotypes is not protective against HEV-C1 infection.
      • An HEV-C1 peptide can be used for specific HEV-C1 serodiagnosis and vaccination.

      Background & Aims

      Rat hepatitis E virus (Orthohepevirus species C; HEV-C1) is an emerging cause of viral hepatitis in humans. HEV-C1 is divergent from other HEV variants infecting humans that belong to Orthohepevirus species A (HEV-A). This study assessed HEV-C1 antigenic divergence from HEV-A and investigated the impact of this divergence on infection susceptibility, serological test sensitivity, and vaccine efficacy.

      Methods

      Immunodominant E2s peptide sequences of HEV-A and HEV-C1 were aligned. Interactions of HEV-C1 E2s and anti-HEV-A monoclonal antibodies (mAbs) were modeled. Recombinant peptides incorporating E2s of HEV-A (HEV-A4 p239) and HEV-C1 (HEV-C1 p241) were expressed. HEV-A and HEV-C1 patient sera were tested using antibody enzymatic immunoassays (EIA), antigen EIAs, and HEV-A4 p239/HEV-C1 p241 immunoblots. Rats immunized with HEV-A1 p239 vaccine (Hecolin), HEV-A4 p239 or HEV-C1 p241 peptides were challenged with a HEV-C1 strain.

      Results

      E2s sequence identity between HEV-A and HEV-C1 was only 48%. There was low conservation at E2s residues (23/53; 43.4%) involved in mAb binding. Anti-HEV-A mAbs bound HEV-C1 poorly in homology modeling and antigen EIAs. Divergence resulted in low sensitivity of commercial antigen (0%) and antibody EIAs (10–70%) for HEV-C1 diagnosis. Species-specific HEV-A4 p239/HEV-C1 p241 immunoblots accurately differentiated HEV-A and HEV-C1 serological profiles in immunized rats (18/18; 100%) and infected-patient sera (32/36; 88.9%). Immunization with Hecolin and HEV-A4 p239 was partially protective while HEV-C1 p241 was fully protective against HEV-C1 infection in rats.

      Conclusions

      Antigenic divergence significantly decreases sensitivity of hepatitis E serodiagnostic assays for HEV-C1 infection. Species-specific immunoblots are useful for diagnosing HEV-C1 and for differentiating the serological profiles of HEV-A and HEV-C1. Prior HEV-A exposure is not protective against HEV-C1. HEV-C1 p241 is an immunogenic vaccine candidate against HEV-C1.

      Lay summary

      Rat hepatitis E virus (HEV-C1) is a new cause of hepatitis in humans. Using a combination of methods, we showed that HEV-C1 is highly divergent from the usual cause of human hepatitis (HEV-A). This divergence reduces the capacity of existing tests to diagnose HEV-C1 and also indicates that prior exposure to HEV-A (via infection or vaccination) is not protective against HEV-C1.

      Graphical abstract

      Keywords

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      References

        • Smith D.B.
        • Simmonds P.
        • International Committee on Taxonomy of Viruses Hepeviridae Study G
        • Jameel S.
        • Emerson S.U.
        • Harrison T.J.
        • Meng X.J.
        • et al.
        Consensus proposals for classification of the family Hepeviridae.
        J Gen Virol. 2014; 95: 2223-2232
        • Sridhar S.
        • Teng J.L.L.
        • Chiu T.H.
        • Lau S.K.P.
        • Woo P.C.Y.
        Hepatitis E virus genotypes and evolution: emergence of camel hepatitis E variants.
        Int J Mol Sci. 2017; 18
        • Pallerla S.R.
        • Harms D.
        • Johne R.
        • Todt D.
        • Steinmann E.
        • Schemmerer M.
        • et al.
        Hepatitis E virus infection: circulation, molecular epidemiology, and impact on global Health.
        Pathogens. 2020; 9
        • Kamar N.
        • Selves J.
        • Mansuy J.M.
        • Ouezzani L.
        • Peron J.M.
        • Guitard J.
        • et al.
        Hepatitis E virus and chronic hepatitis in organ-transplant recipients.
        N Engl J Med. 2008; 358: 811-817
        • Sridhar S.
        • Chan J.F.W.
        • Yap D.Y.H.
        • Teng J.L.L.
        • Huang C.
        • Yip C.C.Y.
        • et al.
        Genotype 4 hepatitis E virus is a cause of chronic hepatitis in renal transplant recipients in Hong Kong.
        J Viral Hepat. 2018; 25: 209-213https://doi.org/10.1111/jvh.12799
        • Zhang J.
        • Zhang X.F.
        • Huang S.J.
        • Wu T.
        • Hu Y.M.
        • Wang Z.Z.
        • et al.
        Long-term efficacy of a hepatitis E vaccine.
        N Engl J Med. 2015; 372: 914-922
        • Wang B.
        • Harms D.
        • Yang X.L.
        • Bock C.T.
        Orthohepevirus C: An expanding species of emerging hepatitis E virus variants.
        Pathogens. 2020; 9
        • Purcell R.H.
        • Engle R.E.
        • Rood M.P.
        • Kabrane-Lazizi Y.
        • Nguyen H.T.
        • Govindarajan S.
        • et al.
        Hepatitis E virus in rats, Los Angeles, California, USA.
        Emerg Infect Dis. 2011; 17: 2216-2222
        • Shimizu K.
        • Hamaguchi S.
        • Ngo C.C.
        • Li T.C.
        • Ando S.
        • Yoshimatsu K.
        • et al.
        Serological evidence of infection with rodent-borne hepatitis E virus HEV-C1 or antigenically related virus in humans.
        J Vet Med Sci. 2016; 78: 1677-1681
        • Dremsek P.
        • Wenzel J.J.
        • Johne R.
        • Ziller M.
        • Hofmann J.
        • Groschup M.H.
        • et al.
        Seroprevalence study in forestry workers from eastern Germany using novel genotype 3- and rat hepatitis E virus-specific immunoglobulin G ELISAs.
        Med Microbiol Immunol. 2012; 201: 189-200
        • Sridhar S.
        • Yip C.C.Y.
        • Wu S.
        • Cai J.
        • Zhang A.J.
        • Leung K.H.
        • et al.
        Rat hepatitis E virus as cause of persistent hepatitis after liver transplant.
        Emerging Infect Dis. 2018; 24: 2241-2250
        • Andonov A.
        • Robbins M.
        • Borlang J.
        • Cao J.
        • Hattchete T.
        • Stueck A.
        • et al.
        Rat hepatitis E virus linked to severe acute hepatitis in an immunocompetent patient.
        J Infect Dis. 2019; 220: 951-955https://doi.org/10.1093/infdis/jiz025
        • Sridhar S.
        • Yip C.C.
        • Wu S.
        • Chew N.F.
        • Leung K.H.
        • Chan J.F.
        • et al.
        Transmission of rat hepatitis E virus infection to humans in Hong Kong: a clinical and epidemiological analysis.
        Hepatology (Baltimore, Md). 2021; 73: 10-22
        • Larkin M.A.
        • Blackshields G.
        • Brown N.P.
        • Chenna R.
        • McGettigan P.A.
        • McWilliam H.
        • et al.
        Clustal W and clustal X version 2.0.
        Bioinformatics. 2007; 23: 2947-2948
        • Zhao M.
        • Li X.J.
        • Tang Z.M.
        • Yang F.
        • Wang S.L.
        • Cai W.
        • et al.
        A comprehensive study of neutralizing antigenic sites on the hepatitis E virus (HEV) capsid by constructing, clustering, and characterizing a tool box.
        J Biol Chem. 2015; 290: 19910-19922
        • Gu Y.
        • Tang X.
        • Zhang X.
        • Song C.
        • Zheng M.
        • Wang K.
        • et al.
        Structural basis for the neutralization of hepatitis E virus by a cross-genotype antibody.
        Cell Res. 2015; 25: 604-620
        • Wen G.P.
        • Tang Z.M.
        • Yang F.
        • Zhang K.
        • Ji W.F.
        • Cai W.
        • et al.
        A valuable antigen detection method for diagnosis of acute hepatitis E.
        J Clin Microbiol. 2015; 53: 782-788
        • Zhang J.
        • Li S.W.
        • Wu T.
        • Zhao Q.
        • Ng M.H.
        • Xia N.S.
        Hepatitis E virus: neutralizing sites, diagnosis, and protective immunity.
        Rev Med Virol. 2012; 22: 339-349
        • Li S.W.
        • Zhang J.
        • Li Y.M.
        • Ou S.H.
        • Huang G.Y.
        • He Z.Q.
        • et al.
        A bacterially expressed particulate hepatitis E vaccine: antigenicity, immunogenicity and protectivity on primates.
        Vaccine. 2005; 23: 2893-2901
        • Debing Y.
        • Mishra N.
        • Verbeken E.
        • Ramaekers K.
        • Dallmeier K.
        • Neyts J.
        A rat model for hepatitis E virus.
        Dis Model Mech. 2016; 9: 1203-1210
        • Kobayashi T.
        • Takahashi M.
        • Tanggis
        • Mulyanto
        • Jirintai S.
        • Nagashima S.
        • et al.
        Characterization and epitope mapping of monoclonal antibodies raised against rat hepatitis E virus capsid protein: an evaluation of their neutralizing activity in a cell culture system.
        J Virol Methods. 2016; 233: 78-88
        • Tsoi W.C.
        • Zhu X.
        • To A.P.
        • Holmberg J.
        Hepatitis E virus infection in Hong Kong blood donors.
        Vox Sang. 2020; 115: 11-17
        • Jirintai S.
        • Tanggis
        • Mulyanto
        • Suparyatmo J.B.
        • Takahashi M.
        • Kobayashi T.
        • et al.
        Rat hepatitis E virus derived from wild rats (Rattus rattus) propagates efficiently in human hepatoma cell lines.
        Virus Res. 2014; 185: 92-102
        • Sanford B.J.
        • Opriessnig T.
        • Kenney S.P.
        • Dryman B.A.
        • Cordoba L.
        • Meng X.J.
        Assessment of the cross-protective capability of recombinant capsid proteins derived from pig, rat, and avian hepatitis E viruses (HEV) against challenge with a genotype 3 HEV in pigs.
        Vaccine. 2012; 30: 6249-6255