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Fig. 5 | Arthritis Research & Therapy

Fig. 5

From: Modulation of B cell activation by extracellular vesicles and potential alteration of this pathway in patients with rheumatoid arthritis

Fig. 5

m/lEVs can directly increase B cell production of antibodies and indirectly activate autologous B cells from patients with RA by innate immune cells. A Representative contour plot of plasmablast population CD27+CD38hi in enriched B cells cultured with CD40L and IL-21 for 7 days. Plasmablasts were analyzed inside of lymphocyte, live/dead negative, and CD19 + regions. B Frequency of plasmablast cells as explained in A, in the absence or presence of HD-m/lEVs, PRA-m/lEVs, or PRA-m/lEVs-ICs. Data of five HD, six patients with RA, and median are shown. C, D Levels of C IgM and D IgG in supernatant of culture of B cells as explained in A, in the absence or presence of HD-m/lEVs, PRA-m/lEVs, or PRA-m/lEVs-ICs. Line indicates lowest detection limit of the ELISA (2 ng/mL). E Monocyte-derived macrophages (MDM) from HD and patients with RA were exposed or not to PRA-m/lEVs or PRA-m/lEVs-ICs for 6 h. Autologous B cells were added to previously washed MDM (B cells: MDM ratio of 1-2:1) and incubated for 96 h. F Frequency of CD69+, CD80+, and CD86.+ B cells from cocultures with MDM unstimulated (Unst) or treated with PRA-m/lEVs or PRA-m/lEV-ICs, as detailed in E. G MDM from patients with RA were exposed or not to PRA-m/lEV-ICs for 6 h. Autologous B cells were cultured with the supernatant of exposed MDM and incubated for 96 h. H Frequency of CD69+, CD80+, and CD86+ B cells cultured with the supernatant of MDM unstimulated (Unst) or treated with PRA-m/lEV-ICs, as detailed in G. Data of six HD and six patients with RA are shown. B–D, F Two-way ANOVA test with Šidák post-test. H Wilcoxon test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

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