Related Experiment Video
Updated: Sep 9, 2025
Human In Vitro Suppression as Screening Tool for the Recognition of an Early State of Immune Imbalance
Published on: July 22, 2011
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.
Insights
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.
Abstract:
Regulatory T cells (Tregs) are essential for maintaining immune balance by controlling the activation and expansion of other immune cells. Conventional suppression assays often rely on co-culturing purified cell populations, which limits multiplexed phenotyping and physiological relevance. This protocol describes a high-dimensional, single-cell assay for profiling Treg-mediated suppression within a peripheral blood mononuclear cell (PBMC) system. Tregs are first isolated by cell sorting and then reintroduced into autologous PBMCs at defined ratios. A 52-marker mass cytometry (CyTOF) panel is used to quantify cell division and phenotypic responses across multiple immune subsets. This approach allows for integrated analysis of Treg function with broad compatibility for patient profiling and drug evaluation. Key features • Quantifies Treg-mediated suppression in autologous PBMCs at single-cell resolution. • Enables high-dimensional phenotyping and proliferation tracking across multiple immune subsets using a 52-marker CyTOF panel. • Maintains physiological relevance by assessing suppression in a complex PBMC environment. • Compatible with patient-derived samples and drug perturbation experiments for translational immunology applications.
