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.
View: Text | PDF
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

×

Figure 2

Antiviral effect of TCR-grafted T cells on HBV-infected cells.

Options: View larger image (or click on image) Download as PowerPoint
Antiviral effect of TCR-grafted T cells on HBV-infected cells. HepG2-NTC...
HepG2-NTCP cells were infected with HBV at a MOI of 100. After 2 weeks, T cells grafted with HBV S–specific TCR 4GS20 (blue squares) or HBV core–specific TCR 6KC18 (red triangles) or nontransduced T cells (mock, gray circles) were added for 10 days at an E/T ratio of 1:2. Medium was changed every other day and used to determine (A) HBeAg and (B) HBsAg by diagnostic ELISA. (C) HBV rcDNA contained in virions that had been secreted was extracted from cell culture supernatant every other day, and DNA extracted from cell lysates on day 10 was used to determine (D) intracellular HBV rcDNA and (E) nuclear cccDNA by qPCR. (F–J) Cells were infected at a MOI of 500. One week after infection, cells were treated with 0.1 μM ETV twice a week for 3 weeks. (F) Killing of target cells was measured using a real-time cell analyzer and is reported as the normalized cell index relative to the starting point of the coculture. E/T of 1:1. (G–J) Medium was changed every 3 to 4 days, and values were normalized for cocultures treated with mock T cells. (G) HBeAg in supernatant of cocultures without or with ETV pretreatment. (H and I) HBV rcDNA contained in virions secreted into the cell culture medium or extracted from cell lysates on day 10, and (J) nuclear cccDNA was determined using qPCR. Data are presented as mean values from triplicate cocultures (n = 3).