ArticleCytometry. Part A : the journal of the International Society for Analytical Cytology2013
A harmonized approach to intracellular cytokine staining gating: Results from an international multiconsortia proficiency panel conducted by the Cancer Immunotherapy Consortium (CIC/CRI).
Article in Cytometry. Part A : the journal of the International Society for Analytical Cytology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers, 1 of them a synthesis that pooled it.
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Who cites it
40 citing papers in PubMed, 1 synthesis or guideline pooled it, 64 citations in OpenAlex.
- Lymphocytes as Biomarkers of Therapeutic Response in Rheumatic Autoimmune Diseases, Is It a Realistic Goal?Clinical reviews in allergy & immunology · 2017Pooled it
- Intracellular Cytokine Staining and Flow Cytometry: Considerations for Application in Clinical Trials of Novel Tuberculosis Vaccines.PloS one · 2015Trial
- Guidelines for Gating Flow Cytometry Data for Immunological Assays.Methods in molecular biology (Clifton, N.J.) · 2027Article
- A decade of innovation in healthcare: Automation, bio-printing and digital twin technologies for personalized therapies.International journal of pharmaceutics: X · 2026Review
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- Automated descriptive cell type naming in flow and mass cytometry with CytoPheno.Scientific reports · 2025Article
- Antigen-specific polyfunctional cytotoxic T cells differentiate intraocular from peripheral blood immune responses in posterior uveitis.Clinical & translational immunology · 2025Article
- Improving Reliability of Immunological Assays by Defining Minimal Criteria for Cell Fitness.ImmunoHorizons · 2024Article
- In Vitro Functionality and Endurance of GMP-Compliant Point-of-Care BCMA.CAR-T Cells at Different Timepoints of Cryopreservation.International journal of molecular sciences · 2024Article
- Antigen-Specific Intraocular Cytokine Responses Distinguish Ocular Tuberculosis From Undifferentiated Uveitis in Tuberculosis-Immunoreactive Patients.American journal of ophthalmology · 2023Article
- Article
- Harmonization and qualification of intracellular cytokine staining to measure influenza-specific CD4Frontiers in immunology · 2022Article
- Adoptive transfer of ex vivo expanded SARS-CoV-2-specific cytotoxic lymphocytes: A viable strategy for COVID-19 immunosuppressed patients?Transplant infectious disease : an official journal of the Transplantation Society · 2021Article
- Unsupervised Analysis of Flow Cytometry Data in a Clinical Setting Captures Cell Diversity and Allows Population Discovery.Frontiers in immunology · 2021Article
- Multicentre Harmonisation of a Six-Colour Flow Cytometry Panel for Naïve/Memory T Cell Immunomonitoring.Journal of immunology research · 2020Article
- Adhering to adhesion: assessing integrin conformation to monitor T cells.Cancer immunology, immunotherapy : CII · 2019Review
- Primary EBV Infection Induces an Acute Wave of Activated Antigen-Specific Cytotoxic CD4Journal of immunology (Baltimore, Md. : 1950) · 2019Article
- Clinical and immunological control of experimental autoimmune encephalomyelitis by tolerogenic dendritic cells loaded with MOG-encoding mRNA.Journal of neuroinflammation · 2019Article
- Data-Driven Flow Cytometry Analysis.Methods in molecular biology (Clifton, N.J.) · 2019Review
- Activated integrins identify functional antigen-specific CD8Proceedings of the National Academy of Sciences of the United States of America · 2018Article
Corrections and comments
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Authors and funding
11 authors at 9 institutions in 2 countries.
Funding
Abstract
Previous results from two proficiency panels of intracellular cytokine staining (ICS) from the Cancer Immunotherapy Consortium and panels from the National Institute of Allergy and Infectious Disease and the Association for Cancer Immunotherapy highlight the variability across laboratories in reported % CD8+ or % CD4+ cytokine-positive cells. One of the main causes of interassay variability in flow cytometry-based assays is due to differences in gating strategies between laboratories, which may prohibit the generation of robust results within single centers and across institutions. To study how gating strategies affect the variation in reported results, a gating panel was organized where all participants analyzed the same set of Flow Cytometry Standard (FCS) files from a four-color ICS assay using their own gating protocol (Phase I) and a gating protocol drafted by consensus from the organizers of the panel (Phase II). Focusing on analysis removed donor, assay, and instrument variation, enabling us to quantify the variability caused by gating alone. One hundred ten participating laboratories applied 110 different gating approaches. This led to high variability in the reported percentage of cytokine-positive cells and consequently in response detection in Phase I. However, variability was dramatically reduced when all laboratories used the same gating strategy (Phase II). Proximity of the cytokine gate to the negative population most impacted true-positive and false-positive response detection. Recommendations are provided for the (1) placement of the cytokine-positive gate, (2) identification of CD4+ CD8+ double-positive T cells, (3) placement of lymphocyte gate, (4) inclusion of dim cells, (5) gate uniformity, and 6) proper adjustment of the biexponential scaling.
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Registered trials
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