OMIP-090: A 20-parameter flow cytometry panel for rapid analysis of cell diversity and homing capacity in human conventional and regulatory T cells
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Updated: Jun 29, 2026
Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
Published on: June 26, 2018
Wladislaw Stroukov1, Daniela Mastronicola1, Caraugh Jane Albany1,2
1"Peter Gorer" Department of Immunobiology, School of Immunology & Microbiological Sciences, King's College London, London, UK.
Insights
This study presents a new flow cytometry panel for analyzing human T cell subsets. It monitors T cell homing and function in fresh blood samples, aiding research in immunology and disease.
Area of Science:
- Immunology
- Cell Biology
- Flow Cytometry
Background:
- Understanding T cell subsets is crucial for immune response analysis.
- Current methods may alter cell integrity due to processing.
- Need for a robust panel to assess T cell homing and function.
Purpose of the Study:
- To develop and optimize a flow cytometry panel for monitoring human CD4+ T cell subsets.
- To assess T cell homing capacity and functional diversity using surface markers.
- To analyze freshly isolated peripheral blood mononuclear cells (PBMCs) to minimize sample alteration.
Main Methods:
- Utilized surface marker expression on freshly isolated PBMCs.
- Included markers for naive/memory populations (CD45RA, CCR7, CD25, CD28, CD95).
- Identified functional subsets using chemokine receptors (CCR4, CCR6, CCR10, CXCR3, CXCR5) and homing markers (CLA, CCR3, CCR5, integrin β7).
Main Results:
- The developed panel effectively monitors CD4+ conventional and regulatory T cell subsets.
- Analysis of T cell distribution (naïve/memory) and functional phenotypes was achieved.
- Homing capacity to various tissues (skin, airway, gut) and inflammatory sites was assessed.
Conclusions:
- The panel provides a reliable method for analyzing T cell subsets without cryopreservation or permeabilization.
- It enables comprehensive assessment of T cell homing and functional diversity.
- The panel is applicable to healthy donors and patients with allergic or autoimmune disorders.
Abstract:
The panel was developed and optimized for monitoring changes in homing capacity and functional diversity of human CD4+ conventional and regulatory T cell subsets. The analysis was based on expression of only surface markers in freshly isolated peripheral blood mononuclear cells (PBMCs) to reduce at minimum any alteration due to permeabilization or freezing/thawing procedures. We included markers to assess the distribution of naïve and memory populations based on the expression of CD45RA, CCR7, CD25, CD28 and CD95 in both conventional and regulatory T cells. The identification of major functional subsets was performed using CCR4, CCR6, CCR10, CXCR3 and CXCR5. Homing capacity of these subsets to skin, airway tract, gut and inflammatory lesions could finally be assessed with the markers CLA, CCR3, CCR5 and integrin β7. The panel was tested on freshly isolated PBMCs from healthy donors and patients with allergic rhinitis or autoimmune disorders.
