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Inclusion of an IgG1-Fc spacer abrogates efficacy of CD19 CAR T cells in a xenograft mouse model

Gene Therapy volume 22, pages 391–403 (2015)Cite this article

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Abstract

Cancer therapy with T cells expressing chimeric antigen receptors (CARs) has produced remarkable clinical responses in recent trials, but also severe side effects. Whereas most protocols use permanently reprogrammed T cells, we have developed a platform for transient CAR expression by mRNA electroporation. This approach may be useful for safe clinical testing of novel receptors, or when a temporary treatment period is desirable. Herein, we investigated therapy with transiently redirected T cells in vitro and in a xenograft mouse model. We constructed a series of CD19-specific CARs with different spacers and co-stimulatory domains (CD28, OX40 or CD28-OX40). The CAR constructs all conferred T cells with potent CD19-specific activity in vitro. Unexpectedly, the constructs incorporating a commonly used IgG1-CH2CH3 spacer showed lack of anti-leukemia activity in vivo and induced severe, partly CD19-independent toxicity. By contrast, identical CAR constructs without the CH2-domain eradicated leukemia in vivo, without notable toxicity. Follow-up studies demonstrated that the CH2CH3-spacer bound soluble mouse Fcγ-receptor I and mediated off-target T-cell activation towards murine macrophages. Our findings highlight the importance of non-signalling CAR elements and of in vivo studies. Finally, the results show that transiently redirected T cells control leukemia in mice and support the rationale for developing an mRNA-CAR platform.

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Acknowledgements

This work was supported by The Norwegian Cancer Society, Radiumhospitalets legater, The Norwegian Health Region South East and the Research Council of Norway. We would like to thank Professor I Sandlie (University of Oslo and OUH) for valuable advice and for providing soluble FcγRs. We also thank Dr AA Tveita (OUH) for providing mouse macrophages and Ms. HJ Hoel, Ms. A Faane and Dr N Westerdaal (OUH) for laboratory assistance. We further thank Dr R Löw (EUFETS AG) for providing the reporter vector and LifeTechnologies for supplying M-450 Epoxy and CD3CD28 Dynabeads. Finally, we would like to thank Dr A Kuhn (BioNTech AG) for valuable advice and Dr C Rössig (University Children's Hospital Münster) and Dr M Pule (University College London) for providing CAR plasmids.

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Author notes

  1. E Walseng and A Kristian: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Oncology, Section for Cell Therapy, Oslo University Hospital Radiumhospitalet, Oslo, Norway

    H Almåsbak, M R Myhre, E M Suso, M Y Wang, G Kvalheim & J A Kyte

  2. Department of Immunology, Oslo University Hospital Radiumhospitalet, Oslo, Norway

    E Walseng & G Gaudernack

  3. Department of Cancer Biology, Scripps Research Institute, Jupiter, FL, USA

    E Walseng

  4. Department of Tumor Biology, Oslo University Hospital Radiumhospitalet, Oslo, Norway

    A Kristian

  5. KG Jebsen Center for Breast Cancer Research, Institute for Clinical Medicine, University of Oslo, Oslo, Norway

    A Kristian

  6. Centre for Immune Regulation and Department of Immunology, Oslo University Hospital Rikshospitalet, Oslo, Norway

    L A Munthe & J T Andersen

  7. Institute of Clinical Medicine, University of Oslo, Oslo, Norway

    L A Munthe

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  1. H Almåsbak
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  2. E Walseng
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Correspondence to J A Kyte.

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Almåsbak, H., Walseng, E., Kristian, A. et al. Inclusion of an IgG1-Fc spacer abrogates efficacy of CD19 CAR T cells in a xenograft mouse model. Gene Ther 22, 391–403 (2015). https://doi.org/10.1038/gt.2015.4

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