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Assessing Human Treg Suppression at Single-Cell Resolution Using Mass Cytometry
Jonas Nørskov Søndergaard1, Janyerkye Tulyeu1, David Priest1
1Human Immunology Team, Center for Infectious Disease Education and Research (CiDER), The University of Osaka, Suita, Japan.
Bio-Protocol
|August 28, 2025
Summary
Regulatory T cells (Tregs) maintain immune balance. This study presents a high-dimensional assay to profile Treg suppression in complex blood cell systems, enhancing immune research and drug evaluation.
Area of Science:
- Immunology
- Cell Biology
- Mass Cytometry
Background:
- Regulatory T cells (Tregs) are crucial for immune homeostasis.
- Current Treg suppression assays lack physiological relevance and multiplexing capabilities.
Purpose of the Study:
- To develop a high-dimensional, single-cell assay for profiling Treg-mediated suppression within autologous peripheral blood mononuclear cells (PBMCs).
- To enable integrated analysis of Treg function and immune responses in a complex cellular environment.
Main Methods:
- Isolation of Tregs via cell sorting.
- Reintroduction of Tregs into autologous PBMCs at defined ratios.
- Utilizing a 52-marker mass cytometry (CyTOF) panel to quantify cell division and phenotypic responses.
Main Results:
- Quantification of Treg-mediated suppression at single-cell resolution within PBMCs.
- High-dimensional phenotyping and proliferation tracking across multiple immune subsets.
- Demonstration of assay compatibility with patient-derived samples and drug perturbation.
Conclusions:
- The developed CyTOF assay provides a physiologically relevant and high-dimensional method for assessing Treg function.
- This approach supports translational immunology research, including patient profiling and drug evaluation.
