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CombiCells allow combinatorial display of cell surface ligands

Nature Reviews Immunology volume 25, page 317 (2025)Cite this article

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The immune and nervous systems need to integrate multiple signals to respond efficiently and appropriately. Although much is known about how cells communicate through soluble ligands such as neurotransmitters and chemokines, which are easy to titrate and use in combination, studying interactions between cell surface ligands is more challenging. One approach to studying these interactions is by genetically modifying cell lines to vary the combination and surface densities of ligands. However, this can require large numbers of cell lines. For example, to study 10 different ligands at 10 different concentrations, you would need more than 100 cell lines subject to genetic drift, which is impractical. Other methods, such as using plate-bound ligands that are immobile or fragile lipid bilayers that are short-lived, also have limitations.

To overcome these challenges, we developed a platform that enables the rapid production of cells with any combination and concentration of ligands. CombiCells are a customizable tool to study how different receptor–ligands interact in a high-throughput manner. With this platform, any cell line, such as CHO-K1, U87 or Nalm6 cells, can be modified to express a SpyCatcher protein that is attached to the transmembrane domain of CD52 on the cell surface. This protein binds to any molecule tagged with a SpyTag, allowing us to decorate a cell surface with any combination and number of ligands in minutes. Depending on the cell type, the ligands stay on the surface for varying amounts of time; for example, 9 h for CHO-K1 cells or over 24 h for Nalm6 cells. We showed that ligands can be combined and titrated to the researcher’s need without outcompeting each other at physiological levels. Researchers can confirm the quantity of ligands on the CombiCell surface by flow cytometry or microscopy.

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References

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  • Patel, A. et al. Using CombiCells, a platform for titration and combinatorial display of cell surface ligands, to study T-cell antigen sensitivity modulation by accessory receptors. EMBO J. 43, 132–150 (2024)

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  • Burton, J. et al. Optimising CAR-T cell sensitivity by engineering matched extracellular sizes between CAR/antigen and CD2/CD58 adhesion complexes. Preprint at bioRxiv https://doi.org/10.1101/2025.01.06.631424 (2025)

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Acknowledgements

CombiCells would not be possible without the efforts in the creation, improvement and application of many researchers from the Dushek and van der Merwe laboratories and collaborators past and present.

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  1. Sir William Dunn School of Pathology, University of Oxford, Oxford, UK

    Sofia Bustamante Eguiguren

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  1. Sofia Bustamante Eguiguren
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Correspondence to Sofia Bustamante Eguiguren.

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The authors declare no competing interests.

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Bustamante Eguiguren, S. CombiCells allow combinatorial display of cell surface ligands. Nat Rev Immunol 25, 317 (2025). https://doi.org/10.1038/s41577-025-01168-z

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