A 2-laser 13-color cFluor ®B Cell Monitoring panel for analyzing human B cell subsets

Abstract

Abstract Flow cytometry is valuable for identifying and monitoring B cell subsets in patients with humoral immune disorders and patients treated with immunotherapies targeting B cells. We have designed a 13-color cFluor ®B Cell Monitoring panel that identifies plasmablasts/plasma cells (CD19 +CD20 −CD27 +CD38 ++), naïve B cells (CD27 −IgD +), IgG class switched memory (CD27 +IgG +) and unswitched memory (CD27 +IgD +and/or IgM +). Additionally, markers for CD4+ and CD8+ T cells are included for a general status of lymphocytes along with granulocyte (CD15) and monocyte (CD14) markers to allow for cleaner lymphocyte gating. The panel was tested on healthy PBMCs and peripheral blood samples collected in EDTA, Cyto-Chex BCT, or heparin tubes. The cell populations of interest were well identified in the PBMCs and blood samples collected in the different types of blood tubes. The resolution of each marker in the panel was comparable to the single-color staining. Evaluation of assay reproducibility showed that the coefficient of variance (CV) for the B cell subsets of interest were <15% in triplicate runs of 10 samples. Also, blood sample stability and post staining stability were evaluated over time. In conclusion, Cytek’s 13-color cFluor ®B Cell Monitoring panel was shown to be effective in identifying and enumerating B cell subsets. This may be a useful tool for monitoring B cell subsets of patients with humoral immune disorders or patients treated with immunotherapies targeting B cells in translational research.