Abstract
T-cell diversity is multifactorial and includes variability in antigen specificity, differentiation, function, and cell-trafficking potential. Spectral overlap limits the ability of traditional flow cytometry to fully capture the diversity of T-cell subsets and function. The development of mass cytometry permits deep immunoprofiling of T-cell subsets, activation state, and function simultaneously from even small volumes of blood. This chapter describes our methods for mass cytometry and high-throughput data analysis of T cells in patient cohorts. We provide a pipeline that includes practical considerations when customizing a panel for mass cytometry. We also provide protocols for the conjugation and titration of metal-labeled antibodies (including two T-cell panels) and a staining procedure. Finally, with the aim to support translational science, we provide R scripts that contain a detailed workflow for initial evaluation of high-dimensional data generated from cohorts of patients.
Key words
- Mass cytometry
- CyTOF
- T cells
- Systems biology
- High-throughput analysis
- R
- Clustering
- Data processing
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Acknowledgements
We thank Divij Matthew, University of Pennsylvania, USA, for his help with the titration figure, as well as Jacob Bergstedt, Lund University, Sweden, for his help with the R scripts.
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O’Boyle, K.C. et al. (2020). Exploration of T-Cell Diversity Using Mass Cytometry. In: Liu, C. (eds) T-Cell Receptor Signaling. Methods in Molecular Biology, vol 2111. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0266-9_1
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DOI: https://doi.org/10.1007/978-1-0716-0266-9_1
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