Lymphocytes transiently expressing virus-specific T cell receptors reduce hepatitis B virus infection
Janine Kah, Sarene Koh, Tassilo Volz, Erica Ceccarello, Lena Allweiss, Marc Lütgehetmann, Antonio Bertoletti, Maura Dandri
Janine Kah, Sarene Koh, Tassilo Volz, Erica Ceccarello, Lena Allweiss, Marc Lütgehetmann, Antonio Bertoletti, Maura Dandri
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Research Article Immunology Infectious disease

Lymphocytes transiently expressing virus-specific T cell receptors reduce hepatitis B virus infection

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.

Authors

Janine Kah, Sarene Koh, Tassilo Volz, Erica Ceccarello, Lena Allweiss, Marc Lütgehetmann, Antonio Bertoletti, Maura Dandri

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Figure 2

mRNA HBV–specific TCR–electroporated T cells show antiviral efficacy in vivo.

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mRNA HBV–specific TCR–electroporated T cells show antiviral efficacy in ...
(A) Schematic representation of the experiment performed to assess the effect of 1 single injection of electroporated effector T cells in high viremic mice reconstituted with haplotype-matched hepatocytes. (B) Viremia changes relative to baseline levels determined after 4 and 6 days in individual mice upon 1 injection of mRNA HBV s183–TCR T cells (n = 5) and in untreated controls (n = 2). (C) ALT levels determined in HBV-infected and uninfected mice receiving a single injection of effector T cells. (D) Schematic representation of the experiment performed to assess the antiviral effect of multiple injections of electroporated effector T cells in high viremic mice reconstituted either with haplotype-matched (n = 3) or -mismatched (n = 3) human hepatocytes and in comparison with mice that were left untreated (n = 3). (E) Median viremia changes determined within each group depicted in D and relative to baseline levels determined in individual mice upon 3 injections of mRNA HBV s183–TCR T cells. Blood was taken 4, 8, and 12 days after the first T cell injection. (F) Median changes in levels of circulating HBeAg were determined by ELISA in all animal groups. BL, baseline.