Systematically defining murine immunity at the phenotypic and functional levels via mass cytometry (P3283)

Abstract

Abstract Despite decades of immunology research in the mouse, no systematic and comprehensive characterization of murine immunity at the proteomic level has been reported. While flow cytometry has been utilized to interrogate immune cells under many circumstances, overlap in fluorochrome emission spectra limits the number of simultaneous parameters available, impeding a thorough analysis of all immune cells and their behavior in complex tissues. Mass cytometry, which couples flow cytometry to time-of-flight mass spectrometry, circumvents this limitation. Here we report the application of single-cell mass cytometry to characterize wild-type murine immunity in three common mouse strains. Using a 40-dimensional panel of immunophenotypic markers, we have assessed the cellular frequency and phenotype of immune cells in the peripheral blood, spleen, lymph nodes, bone marrow, and thymus in an unsupervised manner. Moreover, by combining phenotypic analysis with phospho-specific flow cytometry, we have determined the cell signaling response of immune cells in these tissues to over 20 different stimuli. By characterizing 10 mice per strain, we have defined the ranges of intra-strain and inter-strain variability in immune cell phenotype and behavior. Thus, we report a wild-type murine immune reference map as a resource for the field. We believe this tool will greatly facilitate thorough characterization of pathology, genetic mutations and therapeutic interventions in mouse models.