Stimulation strength controls the rate of initiation but not the molecular organization of TCR-induced signalling

  1. Claire Y Ma
  2. John C Marioni  Is a corresponding author
  3. Gillian M Griffiths  Is a corresponding author
  4. Arianne C Richard  Is a corresponding author
  1. University of Cambridge, United Kingdom
  2. Wellcome Trust Sanger Institute, United Kingdom

Abstract

Millions of naïve T cells with different TCRs may interact with a peptide-MHC ligand, but very few will activate. Remarkably, this fine control is orchestrated using a limited set of intracellular machinery. It remains unclear whether changes in stimulation strength alter the programme of signalling events leading to T cell activation. Using mass cytometry to simultaneously measure multiple signalling pathways during activation of murine CD8+ T cells, we found a programme of distal signalling events that is shared, regardless of the strength of TCR stimulation. Moreover, the relationship between transcription of early response genes Nr4a1 and Irf8 and activation of the ribosomal protein S6 is also conserved across stimuli. Instead, we found that stimulation strength dictates the rate with which cells initiate signalling through this network. These data suggest that TCR-induced signalling results in a coordinated activation program, modulated in rate but not organization by stimulation strength.

Data availability

Raw mass cytometry data can be found on the Flow Repository, accession numbers FR-FCM-Z2CX and FR-FCM-Z2CP.Full results of mass cytometry analyses are included as Supplementary File 5.Source data for summary plots of flow cytometry-measured signaling markers in T cells stimulated with peptide-loaded BMDCs (Figure 7a) are included as Figure 7 - Source Data File 1.Analysis code is available athttps://github.com/MarioniLab/SignallingMassCytoStimStrength

Article and author information

Author details

  1. Claire Y Ma

    Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4244-7535
  2. John C Marioni

    Wellcome Trust Sanger Institute, Hinxton, United Kingdom
    For correspondence
    marioni@ebi.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9092-0852
  3. Gillian M Griffiths

    Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom
    For correspondence
    gg305@cam.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0434-5842
  4. Arianne C Richard

    Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom
    For correspondence
    acr62@cam.ac.uk
    Competing interests
    The authors declare that no competing interests exist.

Funding

Wellcome (103930,100140,217100)

  • Gillian M Griffiths

Wellcome (204017/Z/16/Z)

  • Claire Y Ma

Cancer Research UK (A17197)

  • John C Marioni

Medical Research Council (MR/P014178/1)

  • Arianne C Richard

Addenbrooke's Charitable Trust, Cambridge University Hospitals (23/17 A (ii))

  • Claire Y Ma

European Molecular Biology Organization

  • John C Marioni

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

Ethics

Animal experimentation: Animal experimentation: Experiments were carried out under Project Licence PPL 70/8590. This research has been regulated under the Animals (Scientific Procedures) Act 1986 Amendment Regulations 2012 following ethical review by the University of Cambridge Animal Welfare and Ethical Review Body (AWERB).

Reviewing Editor

  1. Michael L Dustin, University of Oxford, United Kingdom

Version history

  1. Received: November 25, 2019
  2. Accepted: May 14, 2020
  3. Accepted Manuscript published: May 15, 2020 (version 1)
  4. Version of Record published: June 22, 2020 (version 2)

Copyright

© 2020, Ma 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

  • 4,972
    Page views
  • 667
    Downloads
  • 13
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

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. Claire Y Ma
  2. John C Marioni
  3. Gillian M Griffiths
  4. Arianne C Richard
(2020)
Stimulation strength controls the rate of initiation but not the molecular organization of TCR-induced signalling
eLife 9:e53948.
https://doi.org/10.7554/eLife.53948

Share this article

https://doi.org/10.7554/eLife.53948

Further reading

    1. Computational and Systems Biology
    James D Brunner, Nicholas Chia
    Research Article

    The microbial community composition in the human gut has a profound effect on human health. This observation has lead to extensive use of microbiome therapies, including over-the-counter 'probiotic' treatments intended to alter the composition of the microbiome. Despite so much promise and commercial interest, the factors that contribute to the success or failure of microbiome-targeted treatments remain unclear. We investigate the biotic interactions that lead to successful engraftment of a novel bacterial strain introduced to the microbiome as in probiotic treatments. We use pairwise genome-scale metabolic modeling with a generalized resource allocation constraint to build a network of interactions between taxa that appear in an experimental engraftment study. We create induced sub-graphs using the taxa present in individual samples and assess the likelihood of invader engraftment based on network structure. To do so, we use a generalized Lotka-Volterra model, which we show has strong ability to predict if a particular invader or probiotic will successfully engraft into an individual's microbiome. Furthermore, we show that the mechanistic nature of the model is useful for revealing which microbe-microbe interactions potentially drive engraftment.

    1. Computational and Systems Biology
    Tae-Yun Kang, Federico Bocci ... Andre Levchenko
    Research Article

    Angiogenesis is a morphogenic process resulting in the formation of new blood vessels from pre-existing ones, usually in hypoxic micro-environments. The initial steps of angiogenesis depend on robust differentiation of oligopotent endothelial cells into the Tip and Stalk phenotypic cell fates, controlled by NOTCH-dependent cell–cell communication. The dynamics of spatial patterning of this cell fate specification are only partially understood. Here, by combining a controlled experimental angiogenesis model with mathematical and computational analyses, we find that the regular spatial Tip–Stalk cell patterning can undergo an order–disorder transition at a relatively high input level of a pro-angiogenic factor VEGF. The resulting differentiation is robust but temporally unstable for most cells, with only a subset of presumptive Tip cells leading sprout extensions. We further find that sprouts form in a manner maximizing their mutual distance, consistent with a Turing-like model that may depend on local enrichment and depletion of fibronectin. Together, our data suggest that NOTCH signaling mediates a robust way of cell differentiation enabling but not instructing subsequent steps in angiogenic morphogenesis, which may require additional cues and self-organization mechanisms. This analysis can assist in further understanding of cell plasticity underlying angiogenesis and other complex morphogenic processes.