MZB cell differentiation is defective in patients with severe SLE.(A) Flow cytometry dot plots and scatter plots demonstrating reduced frequency of CD27+IgD+ (MZB) cells in a patient with SLE compared with an HCD. (B) Scatter plots of flow cytometry data demonstrating reduced frequency of MZB in patients with SLE compared with HCDs (medians, Mann–Whitney test). ****, P < 0.0001. (C) Scatter plots of flow cytometry data demonstrating reduced MZB frequency in LN, other lupus subtypes (OL) but not in ulcerative colitis (UC), granulomatosis with polyangiitis (GPA), and PV (medians, Mann–Whitney test). ***, P < 0.001; ****, P < 0.0001. (D) Flow cytometry dot plots of a HCD and an SLE patient demonstrating identification of T3 (R123hi) and naive (R123lo) subsets with high and low expression of CD45RB. Stark reduced frequency of CD45RBhi T3 and naive (MZP) populations is evident in SLE. (E) Scatter plot of flow cytometry data demonstrating reduced frequency of MZP (CD45RBhiR123lo) cells in LN patients (medians, Mann–Whitney test). *, P < 0.05; **, P < 0.01; ****, P < 0.0001. (F) Scatter plot of flow cytometry data demonstrating reduced frequency of CD45RBhi T3 (R123hi) cells in LN patients (medians, Mann–Whitney test). ***, P < 0.001. (G) Scatter plot of flow cytometry data demonstrating reduced frequency of naive (CD45RBloR123lo) cells in LN patients (mean ± SD, unpaired t test). *, P < 0.05; **, P < 0.01. (H) Scatter plot of flow cytometry data demonstrating enrichment of CD45RBloR123hi cells in LN patients (mean ± SD, unpaired t test). *, P < 0.05; ***, P < 0.001; ****, P < 0.0001. (I) Gating strategy to distinguish T3 (TS3) and aNAV cell subsets among R123hiCD45RBlo cells on the basis of CD24 and CD38 expression. (J) Scatter plot demonstrating that R123hiCD45RBlo cells were mostly T3 cells as gated in I (mean ± SD). (K) Scatter plot demonstrating increased frequency of T3 cells in OL and LN patients (mean ± SD, unpaired t test). *, P < 0.05; ****, P < 0.0001. (L) Scatter plot demonstrating increased frequency of aNAV cells in OL and LN patients (mean ± SD, unpaired t test). **, P < 0.01. (M) Scatter plots showing the proportion of CD27−IgD− B cells as gated in A demonstrate increased frequency of this population in LN (medians, Mann–Whitney test). *, P < 0.05. (N) Flow cytometry dot plot demonstrating the identification of DN1 and DN2 cells based on expression of CD21 and CD24. (O) Scatter plot showing that DN2 cells as gated in N were more abundant in LN and OL patients than in HCD (medians, Mann–Whitney test). ****, P < 0.0001.
Figure 6.

MZB cell differentiation is defective in patients with severe SLE. (A) Flow cytometry dot plots and scatter plots demonstrating reduced frequency of CD27+IgD+ (MZB) cells in a patient with SLE compared with an HCD. (B) Scatter plots of flow cytometry data demonstrating reduced frequency of MZB in patients with SLE compared with HCDs (medians, Mann–Whitney test). ****, P < 0.0001. (C) Scatter plots of flow cytometry data demonstrating reduced MZB frequency in LN, other lupus subtypes (OL) but not in ulcerative colitis (UC), granulomatosis with polyangiitis (GPA), and PV (medians, Mann–Whitney test). ***, P < 0.001; ****, P < 0.0001. (D) Flow cytometry dot plots of a HCD and an SLE patient demonstrating identification of T3 (R123hi) and naive (R123lo) subsets with high and low expression of CD45RB. Stark reduced frequency of CD45RBhi T3 and naive (MZP) populations is evident in SLE. (E) Scatter plot of flow cytometry data demonstrating reduced frequency of MZP (CD45RBhiR123lo) cells in LN patients (medians, Mann–Whitney test). *, P < 0.05; **, P < 0.01; ****, P < 0.0001. (F) Scatter plot of flow cytometry data demonstrating reduced frequency of CD45RBhi T3 (R123hi) cells in LN patients (medians, Mann–Whitney test). ***, P < 0.001. (G) Scatter plot of flow cytometry data demonstrating reduced frequency of naive (CD45RBloR123lo) cells in LN patients (mean ± SD, unpaired t test). *, P < 0.05; **, P < 0.01. (H) Scatter plot of flow cytometry data demonstrating enrichment of CD45RBloR123hi cells in LN patients (mean ± SD, unpaired t test). *, P < 0.05; ***, P < 0.001; ****, P < 0.0001. (I) Gating strategy to distinguish T3 (TS3) and aNAV cell subsets among R123hiCD45RBlo cells on the basis of CD24 and CD38 expression. (J) Scatter plot demonstrating that R123hiCD45RBlo cells were mostly T3 cells as gated in I (mean ± SD). (K) Scatter plot demonstrating increased frequency of T3 cells in OL and LN patients (mean ± SD, unpaired t test). *, P < 0.05; ****, P < 0.0001. (L) Scatter plot demonstrating increased frequency of aNAV cells in OL and LN patients (mean ± SD, unpaired t test). **, P < 0.01. (M) Scatter plots showing the proportion of CD27−IgD− B cells as gated in A demonstrate increased frequency of this population in LN (medians, Mann–Whitney test). *, P < 0.05. (N) Flow cytometry dot plot demonstrating the identification of DN1 and DN2 cells based on expression of CD21 and CD24. (O) Scatter plot showing that DN2 cells as gated in N were more abundant in LN and OL patients than in HCD (medians, Mann–Whitney test). ****, P < 0.0001.

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