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 1

Genetic engineering and analysis of HBV-specific T cells.

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Genetic engineering and analysis of HBV-specific T cells. (A) Codon-opti...
(A) Codon-optimized TCR α (TRAV) and β (TRBV) chains of high-affinity TCRs were cloned into the retroviral vector MP71. Murine constant domains (mTRBC and mTRAC) and insertion of additional cysteines were used to increase pairing. After retroviral transduction, T cells were stained for mTRBC, and TCR expression was quantified by flow cytometry. (B) A functional comparison scored 11 TCRs directed against the HBV peptides core18–27, S20–28, and S172–180 (21). The TCRs 6KC18 and 4GS20 were identified as having the highest functional avidity and were therefore chosen for further analyses: Parental HepG2 or HBV+ HepG2.2.15 target cells were cocultured with increasing numbers of TCR-grafted T cells. Killing of target cells was determined by detachment from the plate using a real-time cell analyzer (XCelligence) and is given as the normalized cell index relative to the starting point of the coculture. HBV-negative target cells cocultured with TCR-grafted T cells are shown as black lines. HBV-positive target cells cocultured with 6KC18-transduced T cells are shown in red and 4GS20-transduced T cells in blue. Data are presented as mean values of quadruplicate cocultures (n = 4).