Figure 1.
Selective increase of plasmablasts (PBs) during relapse of NMO.
(A) B-cell subpopulation analysis by flow cytometry. Peripheral blood mononuclear cell (PBMC) and cerebrospinal fluid (CSF) cells were obtained during relapse of neuromyelitis optica (NMO) or multiple sclerosis (MS) and were stained with fluorescence-conjugated anti-CD19, -CD27, -CD38, and -CD180 monoclonal antibodies (mAbs). PB cells (CD19intCD27high) were encircled after observing that they also bear the phenotype of CD38highCD180− (Figure S1). Values represent the percentages of PB cells among all mononuclear cells.
(B) The proportion of PB and memory B-cells (mB) in PBMC and CSF from MS and NMO during relapse. The data were obtained from eight patients with MS and five with NMO [*p < 0.05 by Mann–Whitney test; each error bar represents the median ± interquartile range (IQR)].
Figure 2.
Kinetics of CD138+HLA-DR+ PB during relapse of NMO.
(A) Analysis of pooled peripheral blood mononuclear cells (PBMC) data from neuromyelitis optica (NMO) in remission and relapse. The plasmablasts (PBs) were subdivided into four subpopulations by considering the expression of CD138 and HLA-DR. The individual data show the percentages of each PB subpopulation among the total PB [*p < 0.05 by Mann–Whitney test; each error bar represents the median ± interquartile range (IQR)].
(B) Enrichment of CD138+HLA-DR+ PBs in the CSF. The PBMC and CSF cells were obtained from NMO during relapse. The values indicate the percentages of CD138+HLA-DR+ PBs among the total PB. The expression level of CD138 was assessed by mean fluorescence intensity (MFI). Representative data of one out of three different cases are shown.
Figure 3.
CD138+HLA-DR+ plasmablasts (PB) cells are recently differentiated IgG-producing PB.
(A) The effects of influenza vaccination on the frequencies of B-cell subpopulations were analyzed. The frequency of each B-cell subpopulation derived from the peripheral blood of healthy subjects before (pre) and seven days after vaccination (post) is shown. Each line connects the values obtained from a single subject (*p < 0.05 by Wilcoxon signed rank test).
(B) The results of intracellular IgG staining of HLA-DR+ PB (left) and HLA-DR- PBs (right) are shown. The values represent the percentages of IgG-producing cells in each PB subpopulation. Representative data of one out of three individuals are displayed.
Figure 4.
CXCR3 expression on plasmablasts (PBs) correlates with the disease state.
(A) CXCR3 and CXCR4 on PB in neuromyelitis optica (NMO) relapse and remission. Here, we compared the mean fluorescence intensity (MFI) of CXCR3 and CXCR4 expression in the peripheral blood PBs during remission and relapse of NMO. MFI of CXCR3 and CXCR4 expressions on CD138+HLA-DR+ PBs were also analyzed [**p < 0.01 and *p < 0.05 by Mann–Whitney test; each error bar represents the median ± interquartile range (IQR]].
(B) B-cell subpopulations derived from peripheral blood mononuclear cells (PBMC) during disease relapse and remission were analyzed by flow cytometry to investigate the expression of CXCR3 and CXCR4. The values represent the percentages of CXCR3+ or CXCR4+ cells within each B-cell subpopulation. Unstained control of PBMC is indicated by Ctrl. Representative data of at least five patients in each disease state are shown.