PMID- 31203151 OWN - NLM STAT- MEDLINE DCOM- 20201207 LR - 20201214 IS - 1600-0641 (Electronic) IS - 0168-8278 (Linking) VI - 71 IP - 4 DP - 2019 Oct TI - Defining virus-specific CD8+ TCR repertoires for therapeutic regeneration of T cells against chronic hepatitis E. PG - 673-684 LID - S0168-8278(19)30349-6 [pii] LID - 10.1016/j.jhep.2019.06.005 [doi] AB - 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(R)-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-alpha 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. CI - Copyright (c) 2019 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved. FAU - Soon, Chai Fen AU - Soon CF AD - Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany. FAU - Behrendt, Patrick AU - Behrendt P AD - 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. FAU - Todt, Daniel AU - Todt D AD - Department for Molecular and Medical Virology, Ruhr-Universitat Bochum, Bochum, Germany. FAU - Manns, Michael Peter AU - Manns MP AD - Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany. FAU - Wedemeyer, Heiner AU - Wedemeyer H AD - 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. FAU - Sallberg Chen, Margaret AU - Sallberg Chen M AD - Department of Dental Medicine and Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden; Shanghai Tenth People's Hospital, Tongji University, Shanghai, China. FAU - Cornberg, Markus AU - Cornberg M AD - 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. Electronic address: Cornberg.Markus@mh-hannover.de. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190614 PL - Netherlands TA - J Hepatol JT - Journal of hepatology JID - 8503886 RN - 0 (Epitopes, T-Lymphocyte) RN - 0 (HLA-A2 Antigen) RN - 0 (Receptors, Antigen, T-Cell) SB - IM CIN - J Hepatol. 2019 Oct;71(4):648-650. PMID: 31447222 MH - CD8-Positive T-Lymphocytes/*immunology MH - Cells, Cultured MH - Drug Discovery MH - Epitopes, T-Lymphocyte/immunology MH - Genetic Techniques MH - HLA-A2 Antigen/*immunology MH - *Hepatitis E/blood/immunology MH - *Hepatitis E virus/genetics/immunology MH - *Hepatitis, Chronic/immunology/therapy/virology MH - Humans MH - Immunity, Cellular/*immunology MH - Immunotherapy/methods MH - Lymphocyte Activation/immunology MH - *Receptors, Antigen, T-Cell/genetics/immunology OTO - NOTNLM OT - CD8+ T cell receptor OT - Chronic hepatitis E OT - HEV-specific T cell responses OT - Hepatitis E virus OT - Immunotherapy OT - T cell-based therapy OT - Transient redirection EDAT- 2019/06/17 06:00 MHDA- 2020/12/15 06:00 CRDT- 2019/06/17 06:00 PHST- 2018/12/10 00:00 [received] PHST- 2019/05/24 00:00 [revised] PHST- 2019/06/05 00:00 [accepted] PHST- 2019/06/17 06:00 [pubmed] PHST- 2020/12/15 06:00 [medline] PHST- 2019/06/17 06:00 [entrez] AID - S0168-8278(19)30349-6 [pii] AID - 10.1016/j.jhep.2019.06.005 [doi] PST - ppublish SO - J Hepatol. 2019 Oct;71(4):673-684. doi: 10.1016/j.jhep.2019.06.005. Epub 2019 Jun 14.