Experiment Overview

Repository ID: FR-FCM-Z293 Experiment name: Characterization of Phenotypes and Functional Activities of Leukocytes from Rheumatoid Arthritis Patients by Mass Cytometry. MIFlowCyt score: 83.43%
Primary researcher: Nicolas Tchitchek PI/manager: Nicolas Tchitchek Uploaded by: Nicolas Tchitchek
Experiment dates: 2017-09-01 - 2019-09-08 Dataset uploaded: Sep 2019 Last updated: Sep 2019
Keywords: [rheumatoid arthritis] [mass cytometry] [Leukocytes] [TLR stimulation] Manuscripts:
Organizations: CEA, Division of Immuno-Virology, DSV/iMETI, Fontenay-aux-Roses, (France)
Purpose: Rheumatoid arthritis (RA) is the most common autoimmune rheumatic disease and leads to persistent chronic inflammation. The pathophysiology of the disease is complex, involving both adaptive and innate immunity. Among innate immune cells, neutrophils have been rarely studied due to their sensitivity to freezing and their not being collected after Ficoll purification.
Conclusion: We show that RA induces the presence of CD11blow neutrophils (33.7% and 9.2% of neutrophils in RA and controls, respectively) associated with the duration of disease. This population additionally exhibited heterogeneous expression of CD16. We also characterized a CD11ahigh Granzyme Bhigh T-cell population possibly associated with disease activity. There was no difference in cytokine expression after the stimulation of immune cells by TLR agonists between RA and controls.
Comments: Methods: We used mass cytometry to perform a multidimensional phenotypic characterization of immune cells from RA-treated patients, which included the simultaneous study of 33 intra- or extra-cellular markers expressed by leukocytes. We were able to focus our study on innate immune cells, especially neutrophils, due to a specific fixation method before freezing. In addition, blood samples were stimulated or not with various TLR agonists to evaluate whether RA-dependent chronic inflammation can lead to immune-cell exhaustion.
Funding: The IDMIT infrastructure is supported by the French government “Programme d’Investissements d’Avenir” (PIA) under Grant ANR-11-INBS-0008 and grant ANR-10-EQPX-02-01 (FlowCyTech facility). NT was supported by fellowships from the ANRS.
Quality control: Cytometry data were acquired using EQ Four-Element Calibration Beads, normalized using Rachel Finck’s MATLAB normalizer 16 and concatenated using the FCS file concatenation tool (Cytobank).

Experiment variables

· Healthy HEA1_NOSTIM.fcs · HEA1_STIM.fcs · HEA2_NOSTIM.fcs · HEA2_STIM.fcs · HEA3_NOSTIM.fcs · HEA3_STIM.fcs · HEA4_NOSTIM.fcs · HEA4_STIM.fcs · HEA5_NOSTIM.fcs · HEA5_STIM.fcs
· RA PAT1_NOSTIM.fcs · PAT1_STIM.fcs · PAT2_NOSTIM.fcs · PAT2_STIM.fcs · PAT3_NOSTIM.fcs · PAT3_STIM.fcs · PAT4_NOSTIM.fcs · PAT4_STIM.fcs · PAT5_NOSTIM.fcs · PAT5_STIM.fcs · PAT6_NOSTIM.fcs · PAT6_STIM.fcs · PAT7_NOSTIM.fcs · PAT7_STIM.fcs · PAT8_NOSTIM.fcs · PAT8_STIM.fcs · PAT9_NOSTIM.fcs · PAT9_STIM.fcs

· TLR stimulation HEA1_STIM.fcs · HEA2_STIM.fcs · HEA3_STIM.fcs · HEA4_STIM.fcs · HEA5_STIM.fcs · PAT1_STIM.fcs · PAT2_STIM.fcs · PAT3_STIM.fcs · PAT4_STIM.fcs · PAT5_STIM.fcs · PAT6_STIM.fcs · PAT7_STIM.fcs · PAT8_STIM.fcs · PAT9_STIM.fcs
· no stimulation HEA1_NOSTIM.fcs · HEA2_NOSTIM.fcs · HEA3_NOSTIM.fcs · HEA4_NOSTIM.fcs · HEA5_NOSTIM.fcs · PAT1_NOSTIM.fcs · PAT2_NOSTIM.fcs · PAT3_NOSTIM.fcs · PAT4_NOSTIM.fcs · PAT5_NOSTIM.fcs · PAT6_NOSTIM.fcs · PAT7_NOSTIM.fcs · PAT8_NOSTIM.fcs · PAT9_NOSTIM.fcs

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