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

Abstract
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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

Claire Y Ma

Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-4244-7535
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.

"This ORCID iD identifies the author of this article:" 0000-0001-9092-0852
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.

"This ORCID iD identifies the author of this article:" 0000-0003-0434-5842
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).

Copyright

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.