CyTOF enables unprecedented resolution of intracellular readouts to reveal novel functional diversity of human T Cells

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Presentation Time: 03:15 PM - 04:30 PM
Poster Board Number: B624
Abstract ID: 5727

Presenting Author:
Christina Mcconney , PhD Student at Boston Univ. Chobanian & Avedisian Sch. of Med.

Abstract:

Measuring functional signatures at the single-cell level in a cross-lineage manner provides key insights into several facets of immunology, including the plasticity of innate T cells, T/B memory cell diversity following infection/vaccination, and the panoply of activities associated with cell therapy success/failure. To better understand the dynamics of the immune system, flow cytometry phenotyping panels often include intracellular markers of functional potential, such as cytokines, phosphorylation events, and transcription factors. Many of these readouts, including cytokines of the Th2/Treg lineages, have been historically difficult to detect in human cells using flow cytometry. We compared the signal resolution of three 13-parameter panels on a Cytek® Aurora spectral cytometer and the CyTOF® XT mass cytometer. Each panel measured surface and intracellular analytes; fluorescent spectral panels were designed with little/no spectral overlap. PBMC-derived data was analyzed using PhenoGraph clustering and visualized by opt-SNE. Data collected on CyTOF XT™ demonstrated superior signal resolution for a range of intracellular readouts, including cytokines IL-5, IL-10, and IL-13. Also, when better signal/noise enabled more accurate clustering, a higher number of immune cell subsets possessing unique signatures resulted from CyTOF datasets. Overall, cytometry by CyTOF provided clearer understanding of the immuno-diversity present in a sample in the context of unbiased analysis.