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
Graphical Abstract
Keywords
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Article info
Publication history
Published online: June 13, 2019
Accepted:
June 5,
2019
Received in revised form:
May 24,
2019
Received:
December 10,
2018
See Editorial, pages 648–650Identification
Copyright
© 2019 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
