Lighaam, Laura C. Unger, Peter-Paul A. Vredevoogd, David W. Verhoeven, Dorit Vermeulen, Ellen Turksma, Annelies W. ten Brinke, Anja Rispens, Theo Marieke van Ham, S. Image_1_In vitro-Induced Human IL-10+ B Cells Do Not Show a Subset-Defining Marker Signature and Plastically Co-express IL-10 With Pro-Inflammatory Cytokines.tif <p>Regulatory B cells (Breg) have been described as a specific immunological subsets in several mouse models. Identification of a human counterpart has remained troublesome, because unique plasma membrane markers or a defining transcription factor have not been identified. Consequently, human Bregs are still primarily defined by production of IL-10. In this study, we sought to elucidate if in vitro-induced human IL-10 producing B cells are a dedicated immunological subset. Using deep immune profiling by multicolor flow cytometry and t-SNE analysis, we show that the majority of cells induced to produce IL-10 co-express pro-inflammatory cytokines IL-6 and/or TNFα. No combination of markers can be identified to define human IL-10<sup>+</sup>TNFα<sup>−</sup>IL-6<sup>−</sup> B cells and rather point to a general activated B cell phenotype. Strikingly, upon culture and restimulation, a large proportion of formerly IL-10 producing B cells lose IL-10 expression, showing that induced IL-10 production is not a stable trait. The combined features of an activated B cell phenotype, transient IL-10 expression and lack of subset-defining markers suggests that in vitro-induced IL-10 producing B cells are not a dedicated subset of regulatory B cells.</p> immune regulation;Breg;IL-10;IL-6;t-SNE analysis;B cell plasticity;cytokine co-expression;B cell activation 2018-09-05
    https://frontiersin.figshare.com/articles/figure/Image_1_In_vitro-Induced_Human_IL-10_B_Cells_Do_Not_Show_a_Subset-Defining_Marker_Signature_and_Plastically_Co-express_IL-10_With_Pro-Inflammatory_Cytokines_tif/7046279
10.3389/fimmu.2018.01913.s001