Identification of human T-cell receptors with optimal affinity to cancer antigens using antigen-negative humanized mice

Abstract

Identifying T-cell receptors (TCRs) that bind tumor-associated antigens (TAAs) with optimal affinity is a key bottleneck in the development of adoptive T-cell therapy of cancer1. TAAs are unmutated self proteins, and T cells bearing high-affinity TCRs specific for such antigens are commonly deleted in the thymus2. To identify optimal-affinity TCRs, we generated antigen-negative humanized mice with a diverse human TCR repertoire restricted to the human leukocyte antigen (HLA) A*02:01 (ref. 3). These mice were immunized with human TAAs, for which they are not tolerant, allowing induction of CD8+ T cells with optimal-affinity TCRs. We isolate TCRs specific for the cancer/testis (CT) antigen MAGE-A1 (ref. 4) and show that two of them have an anti-tumor effect in vivo. By comparison, human-derived TCRs have lower affinity and do not mediate substantial therapeutic effects. We also identify optimal-affinity TCRs specific for the CT antigen NY-ESO. Our humanized mouse model provides a useful tool for the generation of optimal-affinity TCRs for T-cell therapy.

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Figure 1: Generation of MAGE-A1-specific TCRs in ABabDII mice.
Figure 2: Specificity of ABabDII derived TCRs.
Figure 3: Functional comparison of ABabDII-derived TCRs with a human-derived TCR in vitro and in vivo.
Figure 4: Functional comparison of a NY-ESO157-specific TCR from ABabDII mice with a patient-derived TCR.

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Acknowledgements

The authors thank G. Willimsky for discussion, S. Kupsch and S. Fürl for technical assistance, and I. Hoeft for animal caretaking. C. Linnemann and T. Schumacher kindly provided the NY-ESO-1157/HLA-A2 multimer. This work was supported by the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich TR36) and the Berlin Institute of Health (BIH).

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M.O. planned and performed most experiments, analyzed data and wrote the manuscript. C.L. initiated the project, established new methods, planned and performed experiments, and analyzed data. M.L., X.C. and I.G. planned and performed experiments, and analyzed data. P.v.d.B. and D.J.S. provided unique reagents and advised on their use. W.U. planned experiments and analyzed data. T.B. supervised the project, analyzed data and wrote the manuscript. All authors revised the manuscript.

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Correspondence to Thomas Blankenstein.

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The Max-Delbrück-Center for Molecular Medicine (T.B., M.O., C.L.) applied for a patent on the MAGE-A1-specific TCRs.

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Obenaus, M., Leitão, C., Leisegang, M. et al. Identification of human T-cell receptors with optimal affinity to cancer antigens using antigen-negative humanized mice. Nat Biotechnol 33, 402–407 (2015). https://doi.org/10.1038/nbt.3147

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