Lymphocytes Transiently Expressing Virus-Specific T Cell Receptors Reduce Hepatitis B Virus Infection

J Clin Invest. 2017 Aug 1;127(8):3177-3188. doi: 10.1172/JCI93024. Epub 2017 Jul 24.

Abstract

Adoptive transfer of T cells engineered to express a hepatitis B virus-specific (HBV-specific) T cell receptor (TCR) may supplement HBV-specific immune responses in chronic HBV patients and facilitate HBV control. However, the risk of triggering unrestrained proliferation of permanently engineered T cells raises safety concerns that have hampered testing of this approach in patients. The aim of the present study was to generate T cells that transiently express HBV-specific TCRs using mRNA electroporation and to assess their antiviral and pathogenetic activity in vitro and in HBV-infected human liver chimeric mice. We assessed virological and gene-expression changes using quantitative reverse-transcriptase PCR (qRT-PCR), immunofluorescence, and Luminex technology. HBV-specific T cells lysed HBV-producing hepatoma cells in vitro. In vivo, 3 injections of HBV-specific T cells caused progressive viremia reduction within 12 days of treatment in animals reconstituted with haplotype-matched hepatocytes, whereas viremia remained stable in mice receiving irrelevant T cells redirected toward hepatitis C virus-specific TCRs. Notably, increases in alanine aminotransferase levels, apoptotic markers, and human inflammatory cytokines returned to pretreatment levels within 9 days after the last injection. T cell transfer did not trigger inflammation in uninfected mice. These data support the feasibility of using mRNA electroporation to engineer HBV TCR-redirected T cells in patients with chronic HBV infection.

MeSH terms

  • Adoptive Transfer*
  • Alanine Transaminase / metabolism
  • Animals
  • Carcinoma, Hepatocellular / immunology
  • Carcinoma, Hepatocellular / virology
  • Coculture Techniques
  • Electroporation
  • Female
  • Gene Expression Profiling
  • Granzymes / metabolism
  • Haplotypes
  • Hep G2 Cells
  • Hepatitis B / immunology*
  • Hepatitis B / therapy
  • Hepatitis B Surface Antigens / immunology
  • Hepatitis B virus
  • Hepatitis B, Chronic / immunology
  • Hepatitis B, Chronic / therapy
  • Humans
  • Inflammation
  • Interferon-gamma / metabolism
  • Liver / metabolism
  • Liver Neoplasms / immunology
  • Liver Neoplasms / virology
  • Lymphocytes / cytology*
  • Male
  • Mice
  • RNA, Messenger / metabolism
  • Receptors, Antigen, T-Cell / metabolism*
  • T-Lymphocytes / virology

Substances

  • Hepatitis B Surface Antigens
  • RNA, Messenger
  • Receptors, Antigen, T-Cell
  • Interferon-gamma
  • Alanine Transaminase
  • GZMB protein, human
  • Granzymes