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

Authors: Slabik, Constanze; Kalbarczyk, Maja; Danisch, Simon; Zeidler, Reinhard; Klawonn, Frank; Volk, Valery; Krönke, Nicole; Feuerhake, Friedrich; Ferreira de Figueiredo, Constanca; Blasczyk, Rainer; Olbrich, Henning; Theobald, Sebastian J.; Schneider, Andreas; Ganser, Arnold; von Kaisenberg, Constantin; Lienenklaus, Stefan; Bleich, Andre; Hammerschmidt, Wolfgang; Stripecke, Renata

Online: https://linkinghub.elsevier.com/retrieve/pii/S2372770520301224

Issue: Mol Ther Oncolytics. 2020 Aug 8;18:504-524.


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/CD3ζ 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, CD8+gp350CAR-T cells showed proof-of-concept preclinical efficacy against impending EBV+ lymphoproliferation and lymphomagenesis.