Novel functional diversity of human T cells is revealed as a result of the unprecedented resolution of intracellular cytokines, transcription factors and phosphoproteins using Mass Cytometry (CyTOF) 4037

Abstract Description To interrogate immune-system dynamics, flow cytometry panels often include intracellular cytokines and markers of functional potential, such as phosphorylation events and transcription factors. However, many intracellular readouts have been historically difficult to detect in human cells using flow cytometry. To assess the intracellular detection capabilities of mass cytometry, we compared the signal resolution of three antibody clone-matched intracellular panels on a Cytek Aurora spectral cytometer and a CyTOF™ XT mass cytometer. The CyTOF datasets demonstrated superior signal resolution for a range of intracellular readouts compared with fluorescent cytometry. Most strikingly, stimulation-specific IL-10+ and IL-13+ cells were only detected using CyTOF technology. Also, for the majority of phospho- and transcription factors assessed, mass cytometry also provided cleaner signal:noise distinction. The superior resolution achieved using CyTOF technology enabled more accurate population clustering, and a higher number of immune cell subsets possessing unique signatures were found, including clear detection of T regulatory 1 and Tc2 populations. Overall, CyTOF technology uniquely provides a comprehensive picture of the immuno-diversity present in a sample, and this platform can help illuminate swaths of non-inflammatory immune responses that are currently in the dark, revealing new, broad classes of therapeutic targets and approaches to treat chronic inflammatory diseases. Topic Categories Technological Innovations in Immunology (TECH)