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Adoptive immunotherapy with genetically engineered T cells: modification of the IgG1 Fc ‘spacer’ domain in the extracellular moiety of chimeric antigen receptors avoids ‘off-target’ activation and unintended initiation of an innate immune response

Gene Therapy volume 17, pages 1206–1213 (2010)Cite this article

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Abstract

Chimeric antigen receptors (CARs, immunoreceptors) are frequently used to redirect T cells with pre-defined specificity, in particular towards tumour cells for use in adoptive immunotherapy of malignant diseases. Specific targeting is mediated by an extracellularly located antibody-derived binding domain, which is joined to the transmembrane and intracellular CD3ζ moiety for T-cell activation. Stable CAR expression in T cells, however, requires a spacer domain interposed between the binding and the transmembrane domain and which is commonly the constant IgG1 Fc domain. We here revealed that CARs with Fc spacer domain bind to IgG Fc gamma receptors (FcγRs), thereby unintentionally activating innate immune cells, including monocytes and natural killer (NK) cells, which consequently secrete high amounts of pro-inflammatory cytokines. Engineered T cells, on the other hand, are likewise activated by FcγR binding resulting in cytokine secretion and lysis of monocytes and NK cells independently of the redirected specificity. To reduce FcγR binding, we modified the spacer domain without affecting CAR expression and antigen binding. Engineered with the modified CAR, T cells are not activated in presence of FcγR+ cells, thereby minimizing the risk of off-target activation while preserving their redirected targeting specificity.

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Acknowledgements

We thank Birgit Hops and Frank Steiger for technical assistance. Our work was supported by a grant from the Deutsche Krebshilfe, Bonn, the ATTACK consortium funded by the European Community and the Fortune programme of the Medical Faculty, University of Cologne.

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  1. Zentrum für Molekulare Medizin Köln, and Klinik I für Innere Medizin, Uniklinik Köln, Universität zu Köln, Köln, Germany

    A Hombach, A A Hombach & H Abken

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  1. A Hombach
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  2. A A Hombach
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Correspondence to H Abken.

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Hombach, A., Hombach, A. & Abken, H. Adoptive immunotherapy with genetically engineered T cells: modification of the IgG1 Fc ‘spacer’ domain in the extracellular moiety of chimeric antigen receptors avoids ‘off-target’ activation and unintended initiation of an innate immune response. Gene Ther 17, 1206–1213 (2010). https://doi.org/10.1038/gt.2010.91

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