Journal List > J Korean Rheum Assoc > v.14(3) > 1003563

Jhun, Kim, Ju, Park, Chang, and Kim: Characterization of FSChigh Memory B Cells from Patients with Active Systemic Lupus Erythematosus

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.

Conclusion:

Our results suggest that abnormally activated FSChigh memory B cells from active SLE patients might be involved in spontaneous production of autoantibodies and induce transition from inactive to active phase of the patients.

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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).
jkra-14-219f1.tif
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.
jkra-14-219f2a.tif
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.
jkra-14-219f2b.tif
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).
jkra-14-219f3.tif
Table 1.
Frequency of plasma cells from healthy, inactive SLE and active SLE
FSCowCD27++CD19low /FSd™CD19+cells FSChigtCD27++CD19fow /FSChigtCD19+cells
Healthy (n=11) 0.43±0.33∗,∗∗ 13.40±7.39∗,∗∗∗
Inactive SLE (n=15) 1.38±1.38 + 26.27±14.85
Active SLE (n=10) 5.82±8.13 59.93±20.58

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)

Table 2.
Frequency of memory B cells from healthy, inactive SLE and active SLE
FSClowCD27+CD19+ /FSClowCD19+cells FSChigtCD27+CD19+ /FSChigtCD19+cells
Healthy (n=11) 29.23±8.24∗,∗∗ 54.48±9.70∗,∗∗
Inactive SLE (n=15) 17.56±12.49∗∗∗ 35.09± 10.22 +
Active SLE (n=10) 15.20±9.67 21.54±13.79

∗Significant difference between healthy and inactive SLE (p<0.01), ∗∗Significant difference between healthy and active SLE (p=0.002), ∗∗∗No significant difference between inactive and active SLE (p=0.601), + Significant difference between inactive and active SLE (p=0.018)

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