Lymphocytes transiently expressing virus-specific T cell receptors reduce hepatitis B virus infection
Janine Kah, … , Antonio Bertoletti, Maura Dandri
Janine Kah, … , Antonio Bertoletti, Maura Dandri
Published July 24, 2017
Citation Information: J Clin Invest. 2017;127(8):3177-3188. https://doi.org/10.1172/JCI93024.
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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 4

Antiviral and inflammatory events after in vivo multiple injections of different mRNA HBV–specific TCR–electroporated T cells.

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Antiviral and inflammatory events after in vivo multiple injections of d...
(A) Median viremia changes relative to baseline levels were determined as indicated after 4, 8, and 12 days upon multiple injections of mRNA HBV s183–TCR T cells (n = 3), mRNA c18-TCR T cells (n = 3), and mRNA mock TCR T cells (n = 3) as well as in untreated controls (n = 6). (B) Individual reduction (shown as percentages) of viremia relative to baseline levels determined on days 4, 8, and 12 upon transfer of mRNA s183–191 T cells and mRNA c18 T cells. Transcriptional changes of human T cell response–related genes (C, GZMB; E, IFNG) were measured by qRT-PCR and normalized against human housekeeping transcripts. (D) ALT levels were determined in uninfected (n = 3) and HBV-infected mice receiving multiple injections of effector T cells presenting s183 (n = 3), c18 (n = 3), or HCV NS3 as mock control (n = 3) in comparison with HBV-infected control mice (n = 6). (F) Median changes in human IFN-γ serum protein levels were determined by multiplex measurement in HBV-infected (s183 median = 1280 ng/ml or c18 median = 160 ng/ml) as well as uninfected (s183 median = 3.6 ng/ml) mice that received 3 injections of HBV-specific effector T cells relative to HBV-infected control mice (n = 4; median = 0.53 ng/ml).