T cell receptor grafting allows virological control of hepatitis B virus infection
Karin Wisskirchen, … , Maura Dandri, Ulrike Protzer
Karin Wisskirchen, … , Maura Dandri, Ulrike Protzer
Published April 30, 2019
Citation Information: J Clin Invest. 2019;129(7):2932-2945. https://doi.org/10.1172/JCI120228.
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Research Article Immunology Virology

T cell receptor grafting allows virological control of hepatitis B virus infection

Abstract

T cell therapy is a promising means to treat chronic hepatitis B virus (HBV) infection and HBV-associated hepatocellular carcinoma. T cells engineered to express an HBV-specific T cell receptor (TCR) may cure an HBV infection upon adoptive transfer. We investigated the therapeutic potential and safety of T cells stably expressing high-affinity HBV envelope– or core–specific TCRs recognizing European and Asian HLA-A2 subtypes. Both CD8+ and CD4+ T cells from healthy donors and patients with chronic hepatitis B became polyfunctional effector cells when grafted with HBV-specific TCRs and eliminated HBV from infected HepG2-NTCP cell cultures. A single transfer of TCR-grafted T cells into HBV-infected, humanized mice controlled HBV infection, and virological markers declined by 4 to 5 log or below the detection limit. Engineered T cells specifically cleared infected hepatocytes without damaging noninfected cells when, as in a typical clinical setting, only a minority of hepatocytes were infected. Cell death was compensated by hepatocyte proliferation, and alanine amino transferase levels peaking between days 5 and 7 normalized again thereafter. Cotreatment with the entry inhibitor myrcludex B ensured long-term control of HBV infection. Thus, T cells stably transduced with highly functional TCRs have the potential to mediate clearance of HBV-infected cells, causing limited liver injury.

Authors

Karin Wisskirchen, Janine Kah, Antje Malo, Theresa Asen, Tassilo Volz, Lena Allweiss, Jochen M. Wettengel, Marc Lütgehetmann, Stephan Urban, Tanja Bauer, Maura Dandri, Ulrike Protzer

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Figure 5

Antiviral activity of TCR-grafted T cells in HBV-infected humanized mice.

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Antiviral activity of TCR-grafted T cells in HBV-infected humanized mice...
(A) USG mice were repopulated with HLA-A*02–matched PHHs, infected with 1 × 107 HBV virions, followed until a stable viremia was established (weeks 12–14), and injected with 2 × 106 TCR-grafted T cells (1 × 106 with 6KC18 plus 1 × 106 with 4GS20; colored symbols; n = 7) or with equal numbers of mock-treated human T cells (gray circles; n = 4). Four mice were sacrificed within three weeks (short-term follow-up, pink hexagons), and three mice were sacrificed eight weeks (long-term follow-up, purple diamonds) after T cell transfer, respectively. Two of eleven mice received a second dosage of either effector cells or mock cells and were sacrificed on day fifteen (indicated by dashed lines in A–F and crossed dots in G–I). (B) ALT activity and progression of (C) HSA or (D) HBV DNA in sera. (E and F) HBeAg and HBsAg levels were determined by immunoassay. (G–I) Intrahepatic HBV RNA and DNA transcripts were quantified by qPCR. (G) Levels of HBV pgRNA were normalized to human GAPDH (hGAPDH) RNA. (H and I) rcDNA and cccDNA were quantified relative to an HBV plasmid standard curve and normalized to human β–globin (hβ-globin). Each data point or longitudinal line represents 1 mouse. Dotted lines represent the technical cutoff of the respective test. For DNA and RNA analyses, dotted lines indicate the lower limit of detection (LLoD), defined as 35 cycles of RT-PCR for pgRNA and 10 HBV rcDNA or cccDNA copies per 1000 or more human β–globin copies.