Full control of ligand positioning reveals spatial thresholds for T cell receptor triggering

Abstract

Elucidating the rules for receptor triggering in cell–cell and cell–matrix contacts requires precise control of ligand positioning in three dimensions. Here, we use the T cell receptor (TCR) as a model and subject T cells to different geometric arrangements of ligands, using a nanofabricated single-molecule array platform. This comprises monovalent TCR ligands anchored to lithographically patterned nanoparticle clusters surrounded by mobile adhesion molecules on a supported lipid bilayer. The TCR ligand could be co-planar with the supported lipid bilayer (2D), excluding the CD45 transmembrane tyrosine phosphatase, or elevated by 10 nm on solid nanopedestals (3D), allowing closer access of CD45 to engaged TCR. The two configurations resulted in different T cell responses, depending on the lateral spacing between the ligands. These results identify the important contributions of lateral and axial components of ligand positioning and create a more complete foundation for receptor engineering for immunotherapy.

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Fig. 1: TCR triggering by surface-bound ligands.
Fig. 2: Small clusters of 37 ligands are optimal for TCR triggering.
Fig. 3: Ligand spacing has distinct effect on TCR triggering on 2D and 3D arrays.
Fig. 4: T cells sense ligand position locally.
Fig. 5: Ligand position affects T cell spreading.
Fig. 6: Axial positioning of ligand controls CD45 exclusion.

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Acknowledgements

The authors thank S. Curado for coordination and S. Davis for insightful comments. This work was supported primarily by the National Science Foundation under award no. CMMI-1300590, by the National Institutes of Health Common Fund Nanomedicine programme, grants PN2 EY016586, R37 AI043542 and P01 A1080192; and Wellcome Trust and Kennedy Trust for Rheumatology Research PRF 100262Z/12/Z. The Columbia Nano Initiative provided cleanroom and processing facilities. We thank M. Cammer of NYULMC OCR for microscopy and analysis support.

Author information

H.C. carried out nanofabrication. H.C. and D.D. contributed equally to development of the bilayer backfill methodology. H.C. and J.M. contributed equally to the data acquisition and analysis. V.M. provided purified ICAM1. S.J.W., M.L.D. and M.P.S. designed the experiments. H.C., J.M., M.L.D. and S.J.W. interpreted data and wrote the manuscript.

Correspondence to Michael L. Dustin or Shalom J. Wind.

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Supplementary Figures 1–13, Supplementary Tables 1–2, Supplementary References.

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Cai, H., Muller, J., Depoil, D. et al. Full control of ligand positioning reveals spatial thresholds for T cell receptor triggering. Nature Nanotech 13, 610–617 (2018). https://doi.org/10.1038/s41565-018-0113-3

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