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

• Published in: PloS one 2017-08, Vol.12 (8), p.e0182936-e0182936
• Main Authors: Wisskirchen, Karin, Metzger, Kai, Schreiber, Sophia, Asen, Theresa, Weigand, Luise, Dargel, Christina, Witter, Klaus, Kieback, Elisa, Sprinzl, Martin F, Uckert, Wolfgang, Schiemann, Matthias, Busch, Dirk H, Krackhardt, Angela M, Protzer, Ulrike
• Format: Article
• Language: English
• Subjects: Antigens; Avidity; Biology and Life Sciences; Cancer; CD4 antigen; CD4-Positive T-Lymphocytes - immunology; CD8 antigen; CD8-Positive T-Lymphocytes - immunology; Cloning; Coculture Techniques; Cytokines; Deoxyribonucleic acid; DNA; Effector cells; Female; Focus groups; Gastroenterology; Genetic aspects; Genetic modification; Genotypes; Health aspects; Hepatitis; Hepatitis B; Hepatitis B - immunology; Hepatitis B Antigens - immunology; Hepatitis B virus; Hepatitis B virus - genetics; Hepatitis B virus - immunology; Hepatocellular carcinoma; Hepatoma; Histocompatibility antigen HLA; HLA-A2 Antigen - immunology; Humans; Immunity; Immunology; Infections; Interferon; Interleukin 2; Liver cancer; Lymphocytes; Lymphocytes B; Lymphocytes T; Major histocompatibility complex; Male; Medicine and Health Sciences; Middle Aged; Peptides; Physiological aspects; Physiology; Receptors; Receptors, Antigen, T-Cell - immunology; Receptors, Antigen, T-Cell - metabolism; Replication; Research and Analysis Methods; T cell antigen receptors; T cell receptors; T-cell receptor; Therapy; Tumor necrosis factor-α; Viral infections; Viral Proteins - metabolism; Virology; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0182936

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.