1. Immunology and Inflammation

Optogenetic control shows that kinetic proofreading regulates the activity of the T cell receptor

  1. O Sascha Yousefi
  2. Matthias Günther
  3. Maximilian Hörner
  4. Julia Chalupsky
  5. Maximilian Wess
  6. Simon M Brandl
  7. Robert W Smith
  8. Christian Fleck
  9. Tim Kunkel
  10. Matias D Zurbriggen
  11. Thomas Höfer
  12. Wilfried Weber
  13. Wolfgang WA Schamel  Is a corresponding author
  1. University of Freiburg, Germany
  2. German Cancer Research Center (DKFZ), Germany
  3. Wageningen UR, Netherlands
https://doi.org/10.7554/eLife.42475
Share this article
Cite this article
  1. O Sascha Yousefi
  2. Matthias Günther
  3. Maximilian Hörner
  4. Julia Chalupsky
  5. Maximilian Wess
  6. Simon M Brandl
  7. Robert W Smith
  8. Christian Fleck
  9. Tim Kunkel
  10. Matias D Zurbriggen
  11. Thomas Höfer
  12. Wilfried Weber
  13. Wolfgang WA Schamel
(2019)
Optogenetic control shows that kinetic proofreading regulates the activity of the T cell receptor
eLife 8:e42475.
https://doi.org/10.7554/eLife.42475
  2. Download BibTeX
  3. Download .RIS

Abstract

The immune system distinguishes between self and foreign antigens. The kinetic proofreading (KPR) model proposes that T cells discriminate self from foreign ligands by the different ligand binding half-lives to the T cell receptor (TCR). It is challenging to test KPR as the available experimental systems fall short of only altering the binding half-lives and keeping other parameters of the interaction unchanged. We engineered an optogenetic system using the plant photoreceptor phytochrome B (PhyB) as a ligand to selectively control the dynamics of ligand binding to the TCR by light. This opto-ligand-TCR system was combined with the unique property of PhyB to continuously cycle between the binding and non-binding states under red light, with the light intensity determining the cycling rate and thus the binding duration. Mathematical modeling of our experimental datasets showed that indeed the ligand-TCR interaction half-life is the decisive factor for activating downstream TCR signaling, substantiating KPR.

Data availability

All data that were analyzed with the mathematical model are provided in source data files.

Article and author information

Author details

  1. O Sascha Yousefi

    Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5304-729X

  2. Matthias Günther

    Division of Theoretical Systems Biology, German Cancer Research Center (DKFZ), Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  3. Maximilian Hörner

    Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  4. Julia Chalupsky

    Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  5. Maximilian Wess

    Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  6. Simon M Brandl

    Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  7. Robert W Smith

    Laboratory of Systems and Synthetic Biology, Wageningen UR, Wageningen, Netherlands
    Competing interests
    The authors declare that no competing interests exist.

  8. Christian Fleck

    Laboratory of Systems and Synthetic Biology, Wageningen UR, Wageningen, Netherlands
    Competing interests
    The authors declare that no competing interests exist.

  9. Tim Kunkel

    Faculty of Biology, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  10. Matias D Zurbriggen

    Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  11. Thomas Höfer

    Division of Theoretical Systems Biology, German Cancer Research Center (DKFZ), Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  12. Wilfried Weber

    Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  13. Wolfgang WA Schamel

    Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany
    For correspondence
    wolfgang.schamel@biologie.uni-freiburg.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4496-3100

Funding

Deutsche Forschungsgemeinschaft (GSC-4)

  • O Sascha Yousefi
  • Maximilian Hörner

Deutsche Forschungsgemeinschaft (EXC294)

  • Wolfgang WA Schamel

Deutsche Forschungsgemeinschaft (EXC81)

  • Thomas Höfer

Deutsche Forschungsgemeinschaft (EXC2189)

  • Wolfgang WA Schamel

Deutsche Forschungsgemeinschaft (INST 39/899-1 FUGG)

  • Wolfgang WA Schamel

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2019, Yousefi et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 7,774
    views
  • 1,170
    downloads
  • 101
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Citations by DOI

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. O Sascha Yousefi
  2. Matthias Günther
  3. Maximilian Hörner
  4. Julia Chalupsky
  5. Maximilian Wess
  6. Simon M Brandl
  7. Robert W Smith
  8. Christian Fleck
  9. Tim Kunkel
  10. Matias D Zurbriggen
  11. Thomas Höfer
  12. Wilfried Weber
  13. Wolfgang WA Schamel
(2019)
Optogenetic control shows that kinetic proofreading regulates the activity of the T cell receptor
eLife 8:e42475.
https://doi.org/10.7554/eLife.42475

Share this article

https://doi.org/10.7554/eLife.42475

Categories and tags

Research organisms

Further reading

    1. Immunology and Inflammation
    2. Physics of Living Systems

    Light-based tuning of ligand half-life supports kinetic proofreading model of T cell signaling

    Doug K Tischer, Orion David Weiner
    Tools and Resources Updated

    T cells are thought to discriminate self from foreign peptides by converting small differences in ligand binding half-life into large changes in cell signaling. Such a kinetic proofreading model has been difficult to test directly, as existing methods of altering ligand binding half-life also change other potentially important biophysical parameters, most notably the mechanical stability of the receptor-ligand interaction. Here we develop an optogenetic approach to specifically tune the binding half-life of a chimeric antigen receptor without changing other binding parameters and provide direct evidence of kinetic proofreading in T cell signaling. This half-life discrimination is executed in the proximal signaling pathway, downstream of ZAP70 recruitment and upstream of diacylglycerol accumulation. Our methods represent a general tool for temporal and spatial control of T cell signaling and extend the reach of optogenetics to probe pathways where the individual molecular kinetics, rather than the ensemble average, gates downstream signaling.