Experiment Overview

Repository ID: FR-FCM-Z3GZ Experiment name: Fanelli et al. Mass cytometry data MIFlowCyt score: 64.82%
Primary researcher: Marco Romano PI/manager: Marco Romano Uploaded by: Marco Romano
Experiment dates: 2019-02-13 - 2021-03-19 Dataset uploaded: Feb 2021 Last updated: Mar 2021
Keywords: [PD/PD-L1] [mass cytometry] [Tregs] Manuscripts:
Organizations: King's College London, King's College London, London, London (United Kingdom)
Purpose: Effect of PD-L1 engagment on CD4 cells
Conclusion: Programmed death cell receptor 1 (PD-1) is expressed on T cells upon T cell receptor (TCR) stimulation. High levels of expression are found on T cells during chronic infection, on tumour-infiltrating T cells and memory T cells. PD-1 ligand 1 (PD-L1) is expressed in most tumour environments, and its binding to PD-1 on T cells drives them either to apoptosis or conversion into regulatory T cells (Tregs). However, the fact that PD-L1 itself is also expressed on T cells upon activation has been largely neglected. In this study, we demonstrated that cross-linking of PD-L1 on human CD25-depleted CD4+ T cells, in combination with signalling through CD3/TCR induces their conversion into highly suppressive T cells. Furthermore, this effect was most prominent in memory T cells. PD-L1 co-stimulation on T cells resulted in reduced levels of ERK phosphorylation and decreased mTOR signalling. Importantly, T cells from rheumatoid arthritis patients exhibited high basal levels of phosphorylated ERK and following PD-L1 cross-linking ERK signalling failed to be down modulated, making them resistant to Treg conversion. Altogether our results suggest that engagement of PD-L1 on memory T cells could play an important role in resolving inflammatory responses and its failure could contribute to ongoing autoimmunity.
Comments: FCS files of manually gated live cells following normalization
Funding: British Heart Foundation
Quality control: CD and intracellular antigen analysis

Experiment variables

· AntiCD3 D1 AntiCD3_normalized_CD4+.fcs · D2 AntiCD3_normalized_CD4+.fcs · D3 AntiCD3_normalized_CD4+.fcs
· AntiCD3_CD28 D1 AntiCD3-28_normalized_CD4+.fcs · D2 AntiCD3-28_normalized_CD4+.fcs · D3 AntiCD3-28_normalized_CD4+.fcs
· AntiCD3_PD-L1 D1 AntiCD3-PDL1 _normalized_CD4+.fcs · D2 AntiCD3-PDL1_normalized_CD4+.fcs · D3 AntiCD3-PDL1_normalized_CD4+.fcs

· D1 D1 AntiCD3-28_normalized_CD4+.fcs · D1 AntiCD3-PDL1 _normalized_CD4+.fcs · D1 AntiCD3_normalized_CD4+.fcs
· D2 D2 AntiCD3-28_normalized_CD4+.fcs · D2 AntiCD3-PDL1_normalized_CD4+.fcs · D2 AntiCD3_normalized_CD4+.fcs
· D3 D3 AntiCD3-28_normalized_CD4+.fcs · D3 AntiCD3-PDL1_normalized_CD4+.fcs · D3 AntiCD3_normalized_CD4+.fcs

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