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Updated: Aug 8, 2026
Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
Published on: June 26, 2018
Further analysis of interleukin-2 receptor subunit expression on the different human peripheral blood mononuclear
1Unité d'Immunogénétique Cellulaire, Département d'Immunologie, Institut Pasteur, Paris, France.
Insights
Interleukin-2 receptor (IL-2R) component expression varies on peripheral blood mononuclear cell (PBMC) subsets. Proper sample handling is crucial, as improper collection can lead to widespread IL-2R detection, impacting research findings.
Area of Science:
- Immunology
- Cell Biology
Background:
- The interleukin-2 receptor (IL-2R) is crucial for T cell and natural killer (NK) cell function.
- Its three components (IL-2Ralpha, IL-2Rbeta, IL-2Rgamma) mediate IL-2 signaling.
- Previous studies reported variable IL-2R expression, necessitating clarification.
Purpose of the Study:
- To investigate the expression of IL-2R components on distinct peripheral blood mononuclear cell (PBMC) subsets.
- To identify factors influencing IL-2R detection, potentially explaining conflicting research.
Main Methods:
- Flow cytometry analysis of PBMC subsets.
- Isolation of PBMCs from heparinized blood.
- Assessment of IL-2Ralpha, IL-2Rbeta, and IL-2Rgamma expression.
Main Results:
- IL-2R components were largely absent on CD4 T cells and B cells.
- Expression on CD8 T cells and NK cells varied, with IL-2Rbeta and IL-2Rgamma detected on some subsets.
- Monocytes consistently expressed IL-2Rgamma.
- Delayed cell isolation or non-heparin anticoagulants led to widespread IL-2R expression.
- IL-2Rgamma was found intracellularly in lymphocytes, despite absent surface expression.
Conclusions:
- PBMC subset-specific expression of IL-2R components is confirmed.
- Sample collection and processing significantly impact IL-2R detection, explaining prior discrepancies.
- Intracellular IL-2Rgamma suggests post-translational regulation of its membrane transport.
Abstract:
We have investigated the expression of the three components of the interleukin-2 receptor (IL-2Ralpha, IL-2Rbeta, and IL-2Rgamma) on the surface of the various peripheral blood mononuclear cell (PBMC) subsets by flow cytometry analysis. The PBMC were immediately isolated (ficoll) from blood collected on heparin as anticoagulant. The three IL-2R components are absent or only marginally detectable on CD4 T lymphocytes. No expression of the IL-2R chains is found for the B lymphocytes. In most donors, the three chains are not detectable on CD8 T lymphocytes, but for a few of them, IL-2Rbeta or IL-2Rgamma are clearly expressed. CD56 high (IL-2Ralpha+) and CD56 low (IL-2Ralpha-) natural killer (NK) cells express IL-2Rbeta, but not IL-2Rgamma. IL-2Rgamma is expressed by monocytes of all donors although with variable intensity. When blood is collected on other anticoagulants or when cells are isolated 1 day after collection, IL-2Ralpha, IL-2Rbeta, and IL-2Rgamma are largely expressed on the surface of most PBMC. This observation provides a possible explanation for divergent data previously reported on IL-2R expression. Finally, we show that IL-2Rgamma, which is not detectable on the cell surface of lymphocytes, is nevertheless expressed and stored as an intracellular component. This result is in agreement with the constitutive expression of the IL-2Rgamma gene and suggests a specific regulatory mechanism for IL-2Rgamma membrane translocation.
