Human CD83-targeted chimeric antigen receptor T cells prevent and treat graft-versus-host disease
Bishwas Shrestha, … , Brian C. Betts, Marco L. Davila
Bishwas Shrestha, … , Brian C. Betts, Marco L. Davila
Published May 21, 2020
Citation Information: J Clin Invest. 2020;130(9):4652-4662. https://doi.org/10.1172/JCI135754.
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Research Article Oncology

Human CD83-targeted chimeric antigen receptor T cells prevent and treat graft-versus-host disease

Abstract

Graft-versus-host disease (GVHD) remains an important cause of morbidity and mortality after allogeneic hematopoietic cell transplantation (allo-HCT). For decades, GVHD prophylaxis has included calcineurin inhibitors, despite their incomplete efficacy and impairment of graft-versus-leukemia (GVL). Distinct from pharmacologic immune suppression, we have developed what we believe is a novel, human CD83-targeted chimeric antigen receptor (CAR) T cell for GVHD prevention. CD83 is expressed on allo-activated conventional CD4+ T cells (Tconvs) and proinflammatory dendritic cells (DCs), which are both implicated in GVHD pathogenesis. Human CD83 CAR T cells eradicate pathogenic CD83+ target cells, substantially increase the ratio of regulatory T cells (Tregs) to allo-activated Tconvs, and provide durable prevention of xenogeneic GVHD. CD83 CAR T cells are also capable of treating xenogeneic GVHD. We show that human acute myeloid leukemia (AML) expresses CD83 and that myeloid leukemia cell lines are readily killed by CD83 CAR T cells. Human CD83 CAR T cells are a promising cell-based approach to preventing 2 critical complications of allo-HCT — GVHD and relapse. Thus, the use of human CD83 CAR T cells for GVHD prevention and treatment, as well as for targeting CD83+ AML, warrants clinical investigation.

Authors

Bishwas Shrestha, Kelly Walton, Jordan Reff, Elizabeth M. Sagatys, Nhan Tu, Justin Boucher, Gongbo Li, Tayyebb Ghafoor, Martin Felices, Jeffrey S. Miller, Joseph Pidala, Bruce R. Blazar, Claudio Anasetti, Brian C. Betts, Marco L. Davila

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Figure 1

Human CD83-targeted CAR T construct and functional characteristics.

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Human CD83-targeted CAR T construct and functional characteristics. (A) ...
(A) An anti-CD83 single-chain variable fragment is followed by a CD8 hinge and transmembrane domain, as well as a 41BB costimulatory domain and CD3ζ activation domain. The CAR is tagged with a fluorescence reporter at the 3′ end. The CAR reporter gene is cloned into an SFG retroviral vector. (B) Graph shows CAR gene transfer among T cells (mean ± SEM) by expression of the intracellular EGFP reporter whereas mock-transduced cells are EGFP negative and CD83 CAR T cells are EGFP positive. (C) Graph demonstrates the relative amount of CD4+ or CD8+ subsets among the mock-transduced or CD83 CAR T cells at day +7 after production (n = 2–3 independent donor experiments). (D and E) The amount of IFN-γ and IL-2 released by mock-transduced or CD83 CAR T cells after stimulation with CD83+ DCs. (F) CD83 CAR T cells or mock-transduced T cells were cocultured with CD83+ DCs and cytotoxicity was measured on a real-time cell analysis system. The data are presented (mean ± SEM) as the average normalized cell index over time for duplicate wells. Normalized cell index is calculated as cell index at a given time point divided by cell index at the normalized time point, which is day 1 after addition of T cells. One representative experiment of 2 is shown. (G) CD83 CAR T cells or mock-transduced T cells were stimulated by CD83+ DCs and the absolute number of T cells (mean ± SEM) was calculated weekly over a 14-day period. One representative experiment of 2 shown. ANOVA (D–G). ***P = 0.0001–0.001; ****P < 0.0001.