Sarene Koh1, Janine Kah2, Christine Y L Tham3, Ninghan Yang4, Erica Ceccarello5, Adeline Chia6, Margaret Chen7, Atefeh Khakpoor6, Andrea Pavesi8, Anthony T Tan6, Maura Dandri9, Antonio Bertoletti10. 1. Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research (A*STAR), Singapore; Lion TCR Private Limited Singapore, Singapore. Electronic address: sarene.koh@liontcr.com. 2. Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. 3. Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research (A*STAR), Singapore; NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore. 4. Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research (A*STAR), Singapore. 5. Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), Singapore. 6. Emerging Infectious Diseases (EID) Program, Duke-NUS Medical School, Singapore. 7. Department of Dental Medicine, Karolinska Institutet, Sweden. 8. Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), Singapore. 9. Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; German Center for Infection Research, Hamburg-Lübeck-Borstel Partner Site, Germany. 10. Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research (A*STAR), Singapore; Emerging Infectious Diseases (EID) Program, Duke-NUS Medical School, Singapore. Electronic address: antonio@duke-nus.edu.sg.
Abstract
BACKGROUND & AIMS: Strategies to develop virus-specific T cells against hepatic viral infections have been hindered by safety concerns. We engineered nonlytic human T cells to suppress replication of hepatitis B virus (HBV) and hepatitis C virus (HCV) without overt hepatotoxicity and investigated their antiviral activity. METHODS: We electroporated resting T cells or T cells activated by anti-CD3 with mRNAs encoding HBV or HCV-specific T-cell receptors (TCRs) to create 2 populations of TCR-reprogrammed T cells. We tested their ability to suppress HBV or HCV replication without lysis in 2-dimensional and 3-dimensional cultures of HepG2.2.15 cells and HBV-infected HepG2-hNTCP cells. We also injected TCR-reprogrammed resting and activated T cells into HBV-infected urokinase-type plasminogen activator/severe combined immunodeficiency disease/interleukin 2γ mice with humanized livers and measured levels of intrahepatic and serological viral parameters and serum alanine aminotransferase. Livers were collected for analysis of gene expression patterns to determine effects of the TCR-reprogrammed T cells. RESULTS: TCR-reprogrammed resting T cells produced comparable levels of interferon gamma but lower levels of perforin and granzyme than activated T cells and did not lyse HCV- or HBV-infected hepatoma cells. Although T-cell secretion of interferon gamma was required to inhibit HCV replication, the HBV-specific TCR-reprogrammed resting T cells reduced HBV replication also through intracellular activation of apolipoprotein B mRNA editing enzyme, catalytic polypeptide 3 (APOBEC3). The mechanism of APOBEC3 intracellular activation involved temporal expression of lymphotoxin-β receptor ligands on resting T cells after TCR-mediated antigen recognition and activation of lymphotoxin-β receptor in infected cells. CONCLUSIONS: We developed TCR-reprogrammed nonlytic T cells capable of activating APOBEC3 in hepatoma cells and in HBV-infected human hepatocytes in mice, limiting viral infection. These cells with limited hepatotoxicity might be developed for treatment of chronic HBV infection.
BACKGROUND & AIMS: Strategies to develop virus-specific T cells against hepatic viral infections have been hindered by safety concerns. We engineered nonlytic human T cells to suppress replication of hepatitis B virus (HBV) and hepatitis C virus (HCV) without overt hepatotoxicity and investigated their antiviral activity. METHODS: We electroporated resting T cells or T cells activated by anti-CD3 with mRNAs encoding HBV or HCV-specific T-cell receptors (TCRs) to create 2 populations of TCR-reprogrammed T cells. We tested their ability to suppress HBV or HCV replication without lysis in 2-dimensional and 3-dimensional cultures of HepG2.2.15 cells and HBV-infected HepG2-hNTCP cells. We also injected TCR-reprogrammed resting and activated T cells into HBV-infected urokinase-type plasminogen activator/severe combined immunodeficiency disease/interleukin 2γ mice with humanized livers and measured levels of intrahepatic and serological viral parameters and serum alanine aminotransferase. Livers were collected for analysis of gene expression patterns to determine effects of the TCR-reprogrammed T cells. RESULTS: TCR-reprogrammed resting T cells produced comparable levels of interferon gamma but lower levels of perforin and granzyme than activated T cells and did not lyse HCV- or HBV-infected hepatoma cells. Although T-cell secretion of interferon gamma was required to inhibit HCV replication, the HBV-specific TCR-reprogrammed resting T cells reduced HBV replication also through intracellular activation of apolipoprotein B mRNA editing enzyme, catalytic polypeptide 3 (APOBEC3). The mechanism of APOBEC3 intracellular activation involved temporal expression of lymphotoxin-β receptor ligands on resting T cells after TCR-mediated antigen recognition and activation of lymphotoxin-β receptor in infected cells. CONCLUSIONS: We developed TCR-reprogrammed nonlytic T cells capable of activating APOBEC3 in hepatoma cells and in HBV-infectedhuman hepatocytes in mice, limiting viral infection. These cells with limited hepatotoxicity might be developed for treatment of chronic HBV infection.
Authors: Daniela Stadler; Martin Kächele; Alisha N Jones; Julia Hess; Christian Urban; Jessica Schneider; Yuchen Xia; Andreas Oswald; Firat Nebioglu; Romina Bester; Felix Lasitschka; Marc Ringelhan; Chunkyu Ko; Wen-Min Chou; Arie Geerlof; Maarten A van de Klundert; Jochen M Wettengel; Peter Schirmacher; Mathias Heikenwälder; Sabrina Schreiner; Ralf Bartenschlager; Andreas Pichlmair; Michael Sattler; Kristian Unger; Ulrike Protzer Journal: EMBO Rep Date: 2021-05-09 Impact factor: 8.807
Authors: Katie Healy; Anna Pasetto; Michał J Sobkowiak; Chai Fen Soon; Markus Cornberg; Soo Aleman; Margaret Sällberg Chen Journal: Cells Date: 2020-06-16 Impact factor: 6.600