Comprehensive immune phenotyping of human peripheral leukocytes by mass cytometry for monitoring immunomodulatory therapies
|Sabine Baumgart, Anette Peddinghaus, Ursula Schulte-Wrede, Henrik E. Mei, Andreas Grützkau Cytometry PART A, Volume 91, Issue 1, 34–38 (2017)|
PURPOSE: This OMIP-034 (mass cytometry) comprehensively characterizes live human peripheral blood leukocytes from fresh, erythrocyte-depleted whole blood with a single measurement. Different from existing OMIP, it relies on mass cytometry rather than conventional, fluorescence cytometry and thereby combines 26 different markers in a single staining cocktail. The panel has been optimized with respect to marker selection, antibody clones used, pairing of reporter metal, and antibody on the background of isotope mass-dependent machine sensitivity and antigen abundance, avoiding background signals, and signal spill-over. The panel is designed for monitoring patients' leukocytes during immunomodulatory clinical studies in the field of chronic inflammatory, especially autoimmune diseases, but is likely to serve well in different settings, too. This OMIP-034 captures neutrophils, eosinophils, basophils, monocytes, dendritic cells, T and B lymphocytes and NK cells, and their subsets, and contains a selection of cell activation markers (Table 2). It permits leukocyte analyses in their original complexity without influences from density gradient centrifugation or cryopreservation (Table 1). Blank channels were included for the extension of the panel with up to eight markers of interest. The entire protocol takes 2 days, with 60- to 90-min acquisition time to generate data of ∼5 × 105 cell events.
CELL TYPE: Peripheral blood leukocytes, PBMC
MACHINE: Fluidigm CyTOF