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Abstract
Objective:
To determine phenotypic and functional characteristics of memory B cells in patients with systemic lupus erythematosus (SLE).
Methods:
The percentage of memory B cell subsets in peripheral blood mononuclear cells (PBMC) from normal control (n=11), inactive (n=15) and active (n=10) SLE patients was determined by Fluorescence Activated Cell Sorter (FACS). In addition, the activation status of memory B cells was measured by the surface expression of CD86 (B7-2). The production of antibodies to chromatin and dsDNA (IgG and IgM type) by isolated memory B cell subsets was examined by enzyme-linked immunosorbent assay (ELISA).
Results:
In this study, we analyzed 2 subtypes of memory B cells: FSC (Forward Side Scat- ter)low and FSChigh memory B cell. The percentage of both subtypes from active and inactive SLE patients was significantly reduced compared to that of normal controls (p<0.01). In addition, the expression of activation markers, CD86 on FSChigh memory B cells from active SLE patients was higher than those of inactive SLE patients and normal controls (p=0.014). Upon stimulation with CpG and IL-15 in vitro for 8 days, isolated FSChigh memory B cells from active SLE patients revealed augmented production of autoantibodies to chromatin and dsDNA.
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Fig. 1.
Frequency of B cell subsets in human PBMC CD19+, CD27- naïve B cells, CD19+, CD27+ memory B cell, and CD19, CD27++ plasma cells were determined in patients with active and inactive SLE and in healthy controls by flow cytometric analysis using 2 different gates; Lymphocyte gate (FSClow) and monocyte gate (FSChigh).
Fig. 2A.
CD86 expression on the memory, naïve and plasma cells from normal control and inactive lupus patients. one of three independent experiments is shown.
Fig. 2B.
CD86 expression on the memory, naïve and plasma cells from normal control and active lupus patients. one of three independent experiments is shown.
Fig. 3.
Autoantibody production by FSC g memory B cells stimulated with CpG and IL-15 for 8 days in vitro each histogram shows the mean results obtained from each group. ∗(normal healthy supernatant).
Table 1.
Frequency of plasma cells from healthy, inactive SLE and active SLE
∗ Significant difference between healthy and inactive SLE(p<0.01), ∗∗No Significant difference between healthy and active SLE (p=0.065), ∗∗∗Significant difference between healthy and active SLE (p<0.001), +Nosignifi- cant difference between inactive and active SLE (p= 0.119), Significant difference between inactive and active SLE (p< 0.001)
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