Abstract
Adoptive immunotherapy of cancer using chimeric antigen receptor (CAR)-engineered T cells with redirected specificity showed efficacy in recent trials. In preclinical models, ‘second-generation’ CARs with CD28 costimulatory domain in addition to CD3ζ performed superior in redirecting T-cell effector functions and survival. Whereas CD28 costimulation sustains physiological T-cell receptor (TCR)–CD3 activation of naïve T cells, the impact of CD28 cosignalling on the threshold of CAR-mediated activation of pre-stimulated T cells without B7–CD28 recruitment remained unclear. Using CARs of different binding affinities, but same epitope specificity, we demonstrate that CD28 cosignalling neither lowered the antigen threshold nor the binding affinity for redirected T-cell activation. ‘Affinity ceiling’ above which increase in affinity does not increase T-cell activation was not altered. Accordingly, redirected tumor cell killing depended on the binding affinity but was likewise effective for CD3ζ and CD28–CD3ζ CARs. In contrast to CD3ζ, CD28–CD3ζ CAR-driven activation was not increased further by CD28–B7 engagement. However, CD28 cosignalling, which is required for interleukin-2 induction could not be replaced by high-affinity CD3ζ CAR binding or high-density antigen engagement. We conclude that CD28 CAR cosignalling does not alter the activation threshold but redirects T-cell effector functions.
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Abbreviations
- APC:
-
antigen-presenting cell
- CAR:
-
chimeric antigen receptor
- IFN-γ:
-
interferon-γ
- IL-2:
-
interleukin-2
- MHC:
-
major histocompatibility complex
- PE:
-
phycoerythrin
- scFv:
-
single-chain fragment of variable region
- TCR:
-
T-cell receptor
- XTT:
-
2,3-bis(2-methoxy-4-nitro-5-sulphonyl)-5((phenyl-amino)carbonyl)-2H-tetrazolium hydroxide.
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Acknowledgements
We thank Drs GP Adams and JD Marks, UCSF, for providing us with the anti-ErbB2 scFv C6.5 and derivatives thereof, and Dr S Davis, University of Oxford, UK, for helpful suggestions to the manuscript.
This study was supported by grants from the Deutsche Krebshilfe, Bonn, the ATTACK Consortium of the European Union and the Köln Fortune Program of the Medical Faculty of the University of Cologne.
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Chmielewski, M., Hombach, A. & Abken, H. CD28 cosignalling does not affect the activation threshold in a chimeric antigen receptor-redirected T-cell attack. Gene Ther 18, 62–72 (2011). https://doi.org/10.1038/gt.2010.127
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DOI: https://doi.org/10.1038/gt.2010.127
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