Measurement of human immune cells producing the immunosuppressive/cell repair cytokines TGF-b, IL-10 and others achieved using a novel 50-parameter CyTOF intracellular cytokine staining panel 4022

Abstract Description Most research of human immune responses investigates inflammation, with anti-inflammatory processes grossly understudied due to historically insufficient detection via fluorescent cytometry. Anti-inflammatory mediators and cell repair processes remain largely unknown, and this gap hinders discovery of therapeutic targets and potentially misguides drug development. Theorizing that mass cytometry would be a superior technology for detection of non- inflammatory cytokines and ideal for use in downstream multivariate data analyses, we developed a 50-marker CyTOF™ panel with comprehensive profiling of immune subsets and 20-plus intracellular cytokines spanning major functional lineages. PBMC were stimulated, stained and acquired on the CyTOF XT system. Datasets were analyzed using PhenoGraph clustering and visualized with opt-SNE to determine cellular diversity. Our findings show clear detection of IL-5, IL-10, IL-13 and TGF-b-producing cells, found in surprising co-expression patterns with the pro-inflammatory cytokine IFNg. Through use of this CyTOF panel, which provides wider visualization of the functional diversity of human immune cells than has been achieved previously without the necessary sacrifice of signal resolution that occurs with large fluorescent panels, we predict that novel immune regulatory mechanisms will be revealed, which could spur new avenues for both the treatment of chronic inflammation and the augmentation of vaccine responses. Topic Categories Technological Innovations in Immunology (TECH)