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Figure 1 | Arthritis Research & Therapy

Figure 1

From: Polychromatic flow cytometry in evaluating rheumatic disease patients

Figure 1

Gating schemes for the analysis of human B cells. (A) Cell aggregates and dead cells were further removed from the lymphocyte population, and the resulting live CD19+CD3 cells were selected for analysis. The customarily used IgD/CD27 scheme classifies peripheral blood B cells into four core subsets: naïve and transitional (N + T) IgD+CD27 B cells, unswitched memory (UM) IgD+CD27+ B cells, switched memory (SM) IgDCD27+ B cells, and double-negative (DN) IgDCD27 B cells. Rightmost panel: autoreactive 9G4+ B cells concentrate within the naïve compartment. (B) With the additional memory panel specific markers, SM and DN cells both exhibit heterogeneous subpopulations. A great majority of DN cells downregulate the expression of CD24 and CD21, while CD95+ and CXCR3+ cells are more frequently observed in SM cells. (C) MitoTracker Green (MTG) in the transitional panel separates IgD+CD27 N + T cells into MTG resting naïve cells (rN) and MTG+ fraction. The latter can be further subset into early (T1/T2) transitional B cells, late (T3) transitional B cells and activated naïve (aN) B cells based on the CD24/CD38 expression pattern. A sizable IgM-only memory cells can be identified in the SM subset as well as in the DN subset. (D) Plasma cell panel illustrates that IgDCD27++CD38++ cells include CD138 plasmablasts (PB) and CD138+ plasma cells (PC); both subsets are highly proliferative in the peripheral blood. The IgDCD27−/+CD38++ region contains a CD24 fraction that is also highly proliferative and is considered to be a pre-plasmablast subset (Pre-PB). 9G4+ plasmablasts are readily identified from systemic lupus erythematosus patients. FSC, forward scatter; SSC, side scatter.

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