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

Nature Nanotechnology volume 13, pages 610–617 (2018)Cite this article

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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.

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Author notes

  1. Haogang Cai

    Present address: Center for Nanoscale Materials, Argonne National Laboratory, Argonne, IL, USA

Authors and Affiliations

  1. Department of Mechanical Engineering, Columbia University, New York, NY, USA

    Haogang Cai

  2. Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY, USA

    James Muller & Michael L. Dustin

  3. The Kennedy Institute of Rheumatology, Nuffield Department of Orthopedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, Oxford, UK

    David Depoil, Viveka Mayya & Michael L. Dustin

  4. Mechanobiology Institute of Singapore, National University of Singapore, Singapore, Singapore

    Michael P. Sheetz

  5. Department of Biological Sciences, Columbia University, New York, NY, USA

    Michael P. Sheetz

  6. Department of Applied Physics and Applied Mathematics, Columbia University, New York, NY, USA

    Shalom J. Wind

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Contributions

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.

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

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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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