Article | Published:

Biophysical mechanism of T-cell receptor triggering in a reconstituted system

Nature volume 487, pages 6469 (05 July 2012) | Download Citation

Abstract

A T-cell-mediated immune response is initiated by the T-cell receptor (TCR) interacting with peptide-bound major histocompatibility complex (pMHC) on an infected cell. The mechanism by which this interaction triggers intracellular phosphorylation of the TCR, which lacks a kinase domain, remains poorly understood. Here, we have introduced the TCR and associated signalling molecules into a non-immune cell and reconstituted ligand-specific signalling when these cells are conjugated with antigen-presenting cells. We show that signalling requires the differential segregation of a phosphatase and kinase in the plasma membrane. An artificial, chemically controlled receptor system generates the same effect as TCR–pMHC, demonstrating that the binding energy of an extracellular protein–protein interaction can drive the spatial segregation of membrane proteins without a transmembrane conformational change. This general mechanism may extend to other receptors that rely on extrinsic kinases, including, as we demonstrate, chimaeric antigen receptors being developed for cancer immunotherapy.

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Acknowledgements

We thank A. van der Merwe and V. Cerundolo for the 1G4 TCR sequence, A. Weiss for cell lines and advice, C. June for the CD19 CAR construct, N. Stuurman and K. Thorn for microscopy help and members of the Vale laboratory for discussions. R.D.V. is a Howard Hughes Medical Institute investigator and J.R.J. is a fellow of the Jane Coffin Childs Memorial Fund.

Author information

Affiliations

  1. Howard Hughes Medical Institute and Department of Cellular and Molecular Pharmacology, University of California, San Francisco, 600 16th Street, San Francisco, California 94158, USA

    • John R. James
    •  & Ronald D. Vale

Authors

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Contributions

J.R.J. conceived the study, collected the data and conducted the analyses. J.R.J. and R.D.V. designed the experiments and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Ronald D. Vale.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-8 and the legends to Supplementary Movies 1-5.

Videos

  1. 1.

    Supplementary Movie 1

    The movie shows that the pervanadate treatment of reconstituted cells causes ZAP70 membrane recruitment - see Supplementary Information file for full legend.

  2. 2.

    Supplementary Movie 2

    This movies contains a 3D reconstruction of the HEK-1G4:APC conjugate - see Supplementary Information file for full legend.

  3. 3.

    Supplementary Movie 3

    In this movie we see that ZAP70 rapidly translocates to the cell-cell interface on TCR/pMHC binding - see Supplementary Information file for full legend.

  4. 4.

    Supplementary Movie 4

    In this movie shows monitoring ZAP70 recruitment to signalling-competent regions with time - see Supplementary Information file for full legend.

  5. 5.

    Supplementary Movie 5

    This movie shows that chimaeric-antigen receptor triggering can lead to extensive convolution of the HEK plasma membrane- see Supplementary Information file for full legend.

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DOI

https://doi.org/10.1038/nature11220

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