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Defining virus-specific CD8+ TCR repertoires for therapeutic regeneration of T cells against chronic hepatitis E

  • Chai Fen Soon
    Affiliations
    Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany
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  • Patrick Behrendt
    Affiliations
    Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany

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

    Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, Germany
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  • Daniel Todt
    Affiliations
    Department for Molecular and Medical Virology, Ruhr-Universität Bochum, Bochum, Germany
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  • Michael Peter Manns
    Affiliations
    Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany
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  • Heiner Wedemeyer
    Affiliations
    Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany

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

    Department of Gastroenterology and Hepatology, University Clinic Essen, Essen, Germany
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  • Author Footnotes
    † Both authors contributed equally.
    Margaret Sällberg Chen
    Footnotes
    † Both authors contributed equally.
    Affiliations
    Department of Dental Medicine and Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden

    Shanghai Tenth People's Hospital, Tongji University, Shanghai, China
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  • Author Footnotes
    † Both authors contributed equally.
    Markus Cornberg
    Correspondence
    Corresponding author. Address: Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Tel: +49 5115326821.
    Footnotes
    † Both authors contributed equally.
    Affiliations
    Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany

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

    Centre for Individualised Infection Medicine (CIIM), Hannover, Germany

    Helmholtz Centre for Infection Research (HZI), Braunschweig, Germany
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  • Author Footnotes
    † Both authors contributed equally.

      Highlights

      • Identified 2 HEV-specific CD8+ T cell epitopes located at the RNA helicase and RNA-dependent RNA polymerase.
      • Identified and sequenced T cell receptor (TCR) repertoires against HEV.
      • Transfer of engineered TCRs to T cells of chronically infected patients confers HEV-specific immunogenicity and cytotoxicity.

      Background & Aims

      Immunosuppressed patients with chronic hepatitis E virus infection (cHEV), who are ineligible or have failed current treatment with off-label ribavirin, are a potential target population for T cell-based therapy. T cell responses are important for viral control. Herein, we aimed to identify human leukocyte antigen (HLA)-A2 restricted HEV-specific CD8+ T cell epitopes and T cell receptors (TCR) targeting these epitopes, as the basis for a redirected TCR treatment approach for patients with cHEV.

      Methods

      HEV genotype 3 overlapping peptide pools were used to screen HEV-specific CD8+ T cell immune responses in HLA-A2+ patients with acute HEV infection and healthy donors, by intracellular cytokine staining. CD8+ T cells targeting the identified epitopes were sorted for sequencing of the TCR repertoires by next generation sequencing. Messenger RNA encoding these TCRs were introduced into lymphocytes of healthy donors and patients with cHEV through TCR redirection. TCR-engineered lymphocytes were evaluated for Dextramer®-binding capacity, target sensitivity and cytotoxicity against peptide-loaded T2 cells.

      Results

      HEV-specific responses were observed across open reading frame (ORF)1 and ORF2 of the HEV genome in patients with acute resolving HEV infection. HLA-A2-restricted HEV-specific CD8+ T cell epitopes targeting the HEV RNA helicase and RNA-dependent RNA polymerase were selected for functional studies. Introduction of HEV-specific TCRs into lymphocytes of immunocompetent donors and patients with chronic hepatitis E enabled the lymphocytes to bind HEV Dextramers, secrete multiple cytokines and exert cytotoxicity in a target-specific manner.

      Conclusion

      We identified TCRs that target HEV-specific CD8+ T cell epitopes, and characterized their immune properties, which may have clinical potential in future T cell-based therapy.

      Lay summary

      Patients who are immunosuppressed are vulnerable to developing chronic liver disease following infection with hepatitis E virus (HEV). To-date, there is no approved therapy for chronic hepatitis E. Interferon-α and ribavirin are off-label treatment options, but their applications are limited by side effects. Thus, immunotherapy, more specifically T cell-based therapy, may be an alternative approach. We designed T cell receptor-engineered T cells that effectively conferred immune cells, taken from patients with chronic hepatitis E, with the ability to recognize virus-specific epitopes and mediate killing of target cells in vitro.

      Graphical abstract

      Keywords

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