Mass Cytometry Assays for Antigen-specific T Cells using CyTOF

Mass cytometry (CyTOF) is a modified version of flow cytometry that uses heavy metal ion tags as labels instead of fluorophores, with detection by mass spectrometry.  We have used CyTOF to characterize antigen-specific T cells specific for CMV and EBV, in healthy and immunocompromised subjects.  In this presentation, advantages and limitations of the platform will be presented, along with adaptations that make rare cell identification easier, such as pre-enrichment and barcoding.  Novel methods for integrative analysis of the results will also be presented, including SPADE and a sparse clustering algorithm.  These will be presented in the context of what we are learning about the phenotypes and functions of CMV- and EBV-specific T cells in health and disease.

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