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Single-cell RNA sequencing of OT-I CD8+ T cells after stimulation with different affinity ligands

ABSTRACT: Activation of CD8+ T cells depends exquisitely on the affinity of the T cell receptor (TCR) for a peptide MHC (pMHC) ligand complex. Here, we activated OT-I transgenic CD8+ T cells with pure peptide and examined early activation responses by single-cell RNA-sequencing. T cells were activated with the high affinity OT-I cognate peptide (N4=SIINFEKL) for 1, 3 or 6 hours, or with reduced affinity peptides (T4=SIITFEKL and G4=SIIGFEKL) or the non-binding peptide (NP68=ASNENMDAM) for 6 hours. Cells were then sorted into 96-well plates by FACS and RNA was sequenced following an adapted Smart-Seq2 protocol.

INSTRUMENT(S): Illumina HiSeq 4000

ORGANISM(S): Mus musculus

SUBMITTER: Arianne Richard

PROVIDER: E-MTAB-6051 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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T cell cytolytic capacity is independent of initial stimulation strength.

Richard Arianne C AC Lun Aaron T L ATL Lau Winnie W Y WWY Göttgens Berthold B Marioni John C JC Griffiths Gillian M GM

Nature immunology 20180716 8

How cells respond to myriad stimuli with finite signaling machinery is central to immunology. In naive T cells, the inherent effect of ligand strength on activation pathways and endpoints has remained controversial, confounded by environmental fluctuations and intercellular variability within populations. Here we studied how ligand potency affected the activation of CD8<sup>+</sup> T cells in vitro, through the use of genome-wide RNA, multi-dimensional protein and functional measurements in sing ...[more]

PMID: 30013148

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