T cell receptor grafting allows virological control of hepatitis B virus infection
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
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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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 7

Long-term follow-up of mice partially infected with HBV and treated with HBV-specific T cells and an entry inhibitor.

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Long-term follow-up of mice partially infected with HBV and treated with...
(A) USG mice were repopulated with HLA-A*02–matched PHHs and infected with 1 × 107 HBV virions. Viral spreading was stopped at week 5 after infection by administration of MyrB (see Methods) (gray blocks). After 1 week of MyrB application, 2 × 106 TCR-grafted T cells (1 × 106 with 6KC18 plus 1 × 106 with 4GS20, n = 5) or mock T cells (gray triangles, n = 3) were transferred. To address the question of whether mice could be reinfected after treatment with effector T cells, MyrB application was stopped in 2 of 5 mice after 3 weeks (MyrB short-term, pink diamonds, n = 2) and followed up until week 13. MyrB was administered continuously in 3 of 5 mice (MyrB long-term, purple circles, n = 3). Mice were sacrificed at week 2 or 3 during short-term follow-up or at week 13 during long-term follow-up. Time course of ALT activity (B), HSA (C), HBV viremia (D), HBeAg (E), and HBsAg (F) in sera followed until week 3 or 13. (G) pgRNA levels were normalized to human GAPDH RNA. (H and I) rcDNA and cccDNA levels were quantified relative to an HBV plasmid standard curve and normalized to human β–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 LLoD, defined as 35 cycles of RT-PCR for pgRNA and 10 HBV DNA or cccDNA copies per 1000 or more human β–globin copies. Dots below the LLoD symbolize undetectable measurements. (J) Representative staining of liver tissue slides from mice treated with either mock or 4G plus 6K effector T cells. Scale bars: 50 μm.