Flow cytometric analysis of CD39 expression on human peripheral blood leucocyte populations.
Upper Plots: Whole blood was stained with either PE Mouse IgG1, κ Isotype Control (Cat No. 554680, Left Plot) or PE Mouse Anti-Human CD39 antibody (Cat No. 567156/567157, Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat No. 555899). A bivariate pseudocolor density plot showing the correlated expression of CD39 (or Ig Isotype control staining) versus side light-scatter signals (SSC-A) was derived from gated events with the forward and side light-scatter characteristics of viable leucocyte populations.
Lower Plots: Human peripheral blood mononuclear cells (PBMC) were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220) and then stained with FITC Mouse Anti-Human CD4 (Cat. No. 555346/561005/561842), PE-Cy™7 Mouse Anti-Human CD25 (Cat. No. 557741/560920/561405), Alexa Fluor® 647 Mouse Anti-Human CD127 (Cat No. 558598/560905) antibodies, and either PE Mouse IgG1, κ Isotype Control (dashed line histogram) or PE Mouse Anti-Human CD39 (solid line histogram). Bivariate pseudocolor density plots showing the coexpressed levels of CD25 versus CD127 by viable light scatter-gated CD4+ T cells [Left Plot] were further gated to reveal CD39 expression or Ig Isotype control staining [Right Plot] on CD4+CD25+CD127low T cells (ie, cells with a Regulatory T cell immunophenotype) as shown.
Flow cytometric analysis was performed using a BD FACSCelelsta™ or LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.