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CAR-T cells and TRUCKs that recognize an EBNA-3C-derived epitope presented on HLA-B∗35 control Epstein-Barr virus-associated lymphoproliferation

Dragon AC, Zimmermann K, Nerreter T, Sandfort D, Lahrberg J, Kloss S, Kloth C, Mangare C, Bonifacius A, Tischer-Zimmermann S, Blasczyk R, Maecker-Kolhoff B, Uchanska-Ziegler B, Abken H, Schambach A, Hudecek M, Eiz-Vesper B (2020)

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Publication Type: Journal article

Publication year: 2020

Journal

Book Volume: 8

Article Number: e000736

Journal Issue: 2

DOI: 10.1136/jitc-2020-000736

Abstract

Background Immunosuppressive therapy or T-cell depletion in transplant patients can cause uncontrolled growth of Epstein-Barr virus (EBV)-infected B cells resulting in post-transplant lymphoproliferative disease (PTLD). Current treatment options do not distinguish between healthy and malignant B cells and are thereby often limited by severe side effects in the already immunocompromised patients. To specifically target EBV-infected B cells, we developed a novel peptide-selective chimeric antigen receptor (CAR) based on the monoclonal antibody TÜ165 which recognizes an Epstein-Barr nuclear antigen (EBNA)-3C-derived peptide in HLA-B∗35 context in a T-cell receptor (TCR)-like manner. In order to attract additional immune cells to proximity of PTLD cells, based on the TÜ165 CAR, we moreover generated T cells redirected for universal cytokine-mediated killing (TRUCKs), which induce interleukin (IL)-12 release on target contact. Methods TÜ165-based CAR-T cells (CAR-Ts) and TRUCKs with inducible IL-12 expression in an all-in-one construct were generated. Functionality of the engineered cells was assessed in co-cultures with EBNA-3C-peptide-loaded, HLA-B∗35-expressing K562 cells and EBV-infected B cells as PTLD model. IL-12, secreted by TRUCKs on target contact, was further tested for its chemoattractive and activating potential towards monocytes and natural killer (NK) cells. Results After co-cultivation with EBV target cells, TÜ165 CAR-Ts and TRUCKs showed an increased activation marker expression (CD137, CD25) and release of proinflammatory cytokines (interferon-γand tumor necrosis factor-α). Moreover, TÜ165 CAR-Ts and TRUCKs released apoptosis-inducing mediators (granzyme B and perforin) and were capable to specifically lyse EBV-positive target cells. Live cell imaging revealed a specific attraction of TÜ165 CAR-Ts around EBNA-3C-peptide-loaded target cells. Of note, TÜ165 TRUCKs with inducible IL-12 showed highly improved effector functions and additionally led to recruitment of monocyte and NK cell lines. Conclusions Our results demonstrate that TÜ165 CAR-Ts recognize EBV peptide/HLA complexes in a TCR-like manner and thereby allow for recognizing an intracellular EBV target. TÜ165 TRUCKs equipped with inducible IL-12 expression responded even more effectively and released IL-12 recruited additional immune cells which are generally missing in proximity of lymphoproliferation in immunocompromised PTLD patients. This suggests a new and promising strategy to specifically target EBV-infected cells while sparing and mobilizing healthy immune cells and thereby enable control of EBV-associated lymphoproliferation.

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How to cite

APA:

Dragon, A.C., Zimmermann, K., Nerreter, T., Sandfort, D., Lahrberg, J., Kloss, S.,... Eiz-Vesper, B. (2020). CAR-T cells and TRUCKs that recognize an EBNA-3C-derived epitope presented on HLA-B∗35 control Epstein-Barr virus-associated lymphoproliferation. Journal for ImmunoTherapy of Cancer, 8(2). https://doi.org/10.1136/jitc-2020-000736

MLA:

Dragon, Anna Christina, et al. "CAR-T cells and TRUCKs that recognize an EBNA-3C-derived epitope presented on HLA-B∗35 control Epstein-Barr virus-associated lymphoproliferation." Journal for ImmunoTherapy of Cancer 8.2 (2020).

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