Isolation and functional characterization of hepatitis B virus-specific T-cell receptors as new tools for experimental and clinical use

PLoS One. 2017 Aug 8;12(8):e0182936. doi: 10.1371/journal.pone.0182936. eCollection 2017.

Abstract

T-cell therapy of chronic hepatitis B is a novel approach to restore antiviral T-cell immunity and cure the infection. We aimed at identifying T-cell receptors (TCR) with high functional avidity that have the potential to be used for adoptive T-cell therapy. To this end, we cloned HLA-A*02-restricted, hepatitis B virus (HBV)-specific T cells from patients with acute or resolved HBV infection. We isolated 11 envelope- or core-specific TCRs and evaluated them in comprehensive functional analyses. T cells were genetically modified by retroviral transduction to express HBV-specific TCRs. CD8+ as well as CD4+ T cells became effector T cells recognizing even picomolar concentrations of cognate peptide. TCR-transduced T cells were polyfunctional, secreting the cytokines interferon gamma, tumor necrosis factor alpha and interleukin-2, and effectively killed hepatoma cells replicating HBV. Notably, our collection of HBV-specific TCRs recognized peptides derived from HBV genotypes A, B, C and D presented on different HLA-A*02 subtypes common in areas with high HBV prevalence. When co-cultured with HBV-infected cells, TCR-transduced T cells rapidly reduced viral markers within two days. Our unique set of HBV-specific TCRs with different affinities represents an interesting tool for elucidating mechanisms of TCR-MHC interaction and dissecting specific anti-HBV mechanisms exerted by T cells. TCRs with high functional avidity might be suited to redirect T cells for adoptive T-cell therapy of chronic hepatitis B and HBV-induced hepatocellular carcinoma.

MeSH terms

  • CD4-Positive T-Lymphocytes / immunology*
  • CD8-Positive T-Lymphocytes / immunology*
  • Coculture Techniques
  • Female
  • HLA-A2 Antigen / immunology
  • Hepatitis B / immunology
  • Hepatitis B Antigens / immunology
  • Hepatitis B virus / genetics
  • Hepatitis B virus / immunology*
  • Humans
  • Male
  • Middle Aged
  • Receptors, Antigen, T-Cell / immunology*
  • Receptors, Antigen, T-Cell / metabolism
  • Viral Proteins / metabolism

Substances

  • HLA-A*02 antigen
  • HLA-A2 Antigen
  • Hepatitis B Antigens
  • Receptors, Antigen, T-Cell
  • Viral Proteins

Grants and funding

This work was supported by the German research Foundation (DFG, www.dfg.de) via TRR36 to U.P., D.B. and W.U., and the Helmholtz Alliance on Immunotherapy of Cancer and the German Center for Infection Research (DZIF, TTU Hepatitis 05.806, www.dzif.de) to U.P.. K.W. received a Maternity Leave and M.F.S. a Clinical Leave stipend from DZIF.