CAR-T Cells Targeting Epstein-Barr Virus gp350 Validated in a Humanized Mouse Model of EBV Infection and Lymphoproliferative Disease

Slabik, Constanze; Kalbarczyk, Maja; Danisch, Simon; Zeidler, Reinhard; Klawonn, Frank; Volk, Valery; Kroenke, Nicole; Feuerhake, Friedrich; de Figueiredo, Constanca Ferreira; Blasczyk, Rainer; Olbrich, Henning; Theobald, Sebastian J.; Schneider, Andreas; Ganser, Arnold; Kaisenberg, Constantin von; Lienenklaus, Stefan; Bleich, Andre; Hammerschmidt, Wolfgang und Stripecke, Renata (2020): CAR-T Cells Targeting Epstein-Barr Virus gp350 Validated in a Humanized Mouse Model of EBV Infection and Lymphoproliferative Disease. In: Molecular Therapy-Oncolytics, Bd. 18: S. 504-524

Abstract

Epstein-Barr virus (EBV) is a latent and oncogenic human herpesvirus. Lytic viral protein expression plays an important role in EBV-associated malignancies. The EBV envelope glycoprotein 350 (gp350) is expressed abundantly during EBV lytic reactivation and sporadically on the surface of latently infected cells. Here we tested T cells expressing gp350-specific chimeric antigen receptors (CARs) containing scFvs derived from two novel gp350-binding, highly neutralizing monoclonal antibodies. The scFvs were fused to CD28/CD zeta signaling domains in a retroviral vector. The produced gp350CAR-T cells specifically recognized and killed gp350(+) 293T cells in vitro. The best-performing 7A1-gp350CAR-T cells were cytotoxic against the EBV+ B95-8 cell line, showing selectivity against gp350(+) cells. Fully humanized Nod.Rag.Gamma mice transplanted with cord blood CD34(+) cells and infected with the EBV/M81/fLuc lytic strain were monitored dynamically for viral spread. Infected mice recapitulated EBV-induced lymphoproliferation, tumor development, and systemic inflammation. We tested adoptive transfer of autologous CD8(+)gp350CAR-T cells administered protectively or therapeutically. After gp350CAR-T cell therapy, 75% of mice controlled or reduced EBV spread and showed lower frequencies of EBER+ B cell malignant lymphoproliferation, lack of tumor development, and reduced inflammation. In summary, CDegp350CAR-T cells showed proof-of-concept preclinical efficacy against impending EBV+ lymphoproliferation and lymphomagenesis.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Medizin
• Themengebiete: 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
• ISSN: 2372-7705
• Sprache: Englisch