Fig. 8: Inhibition of GSK3 in AR B cell to restore the capability inducing Tr1 cells.

B cells (from AR patients) were primed with CD40L/CpG in the culture in the presence or absence of SB (10 µM) for 24 h, and cultured with naive CD4⁺ T cells (from HC subjects, n = 3, and AR subjects, n = 3) at ratio of 1:10 (B cell:T cell) for 3 days. Cells were then analyzed by FACS. a CD4⁺ T cells were gated. b Gated cells show Tr1 cell counts. c Dot plots show Tr1 cell frequency from three independent experiments. d, e CD4⁺ CD25¯ CD62L⁺ T cells (Naive CD4⁺ T cells; from HC subjects; labeled with CFSE) were cultured with Tr1 cells (generated as described in b) at ratio of 1:10 (Tr1:Teff) in the presence of anti-CD3/CD28 Abs for 3 days. Teff proliferation was assessed by FACS. Gated histograms show proliferating Teff counts. Dot plots show proliferating Teff frequency. ***p < 0.001 (ANOVA followed by the Dunnett’s post hoc test), compared with group a (c) or group b (e). NS no stimulation group. Group labels of c, e are the same as FACS plots on the left side. The data of a, b, d are from one experiment, that represent three independent experiments.