Abstract
Despite significant clinical progress, checkpoint blockade remains limited by variable response rates, resistance, and toxicity. Activating costimulatory receptors offers a promising alternative to enhance anti-tumor immunity. However, there is insufficient understanding of how to mimic physiological membrane-anchored costimulatory ligands. Here, we describe a strategy for developing effective agonists of the costimulatory receptor CD27 by increasing both antibody valency and FcγRIIB engagement. Engineered anti-CD27 antibodies capable of tetravalent binding to CD27 and selective FcγRIIB association exhibit potent T cell stimulatory activity and anti-tumor efficacy in pre-clinical models, compared to bivalent counterparts. The anti-tumor effects of the tetravalent antibody are mediated through CD8⁺ T cell activation without evidence of regulatory T cell depletion. Mechanistically, whereas the increase in avidity drives more efficient CD27 clustering, FcγRIIB engagement triggers polarization of receptor clusters to the cell-cell interface and reduces receptor internalization. This work provides a framework for developing more effective agonist-based T cell stimulatory therapies.
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All data are included in the Supplementary Information or available from the authors, as are unique reagents used in this article. The raw numbers for charts and graphs are available in the Source data file whenever possible. Source data are provided with this paper.
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Acknowledgements
The authors thank Dr Hannah Smith and Dr Robert Oldham for the kind gift of CHO cells expressing FcγRs. The authors would like to acknowledge that the purchase of the Nanoimager used in the microscopy studies was made possible by a kind gift from the Mark Benevolent Fund. The authors are grateful to members of the Biomedical Research Facility, University of Southampton, for their help with the murine in vivo studies. The study was funded by Cancer Research UK Award number DRCDDRPGMApr2020\100005 (A.Al-S., S.A.B., M.S.C.).
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M.A.W. performed and analyzed the experiments with the help of A.P., H.J.M., S.G.B., H.T.C.C., T.I., C.A.P., C.I.M., and S.J. A.Al-S. conceived the project. A.Al-S., S.A.B., and M.S.C. supervised the project. M.A.W. and A.Al-S. wrote the manuscript with feedback from M.S.C., S.A.B., and S.H.L.
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A.Al-S. and S.H.L. are inventors on patents pertaining to the generation and therapeutic use of agonist anti-CD27 antibodies and have received research funding from Celldex Therapeutics in relation to anti-CD27 antibodies. The remaining authors declare no competing interests.
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Widdess, M.A., Pakidi, A., Metcalfe, H.J. et al. Harnessing multivalency and FcγRIIB engagement to augment anti-CD27 immunotherapy. Nat Commun (2025). https://doi.org/10.1038/s41467-025-67882-3
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DOI: https://doi.org/10.1038/s41467-025-67882-3
