Modulating Oxidative Stress in B Cells Promotes Immunotherapy in Food Allergy

Figure 3

FGN counteracts the specific antigen-induced Breg apoptosis. LPMCs were prepared from naïve BALB/c mice or mice treated by gavage-feeding with OVA daily for one week and exposed to OVA (or BSA) in the culture overnight. (a) LPMCs were analyzed by FACS. Gated plots show GL7⁺ B cells (activated B cells). (b) Gated plots show CD5⁺ IL-10⁺ cells in the gated cells in panel Ad. (c) Boxplots show activated (by OVA) B cell frequency. (d–g) CD5⁺ B cells were isolated from LPMCs prepared naïve mice and OVA-primed mice and cultured in the presence of OVA (or BSA) overnight. Cells were stained with propidium iodide and Annexin V reagent and analyzed by FACS. Gated plots show (d) apoptotic cells. Boxplots show apoptotic (e) Breg frequency or (f) FasL mRNA in Breg. Immunoblots show (g) FasL protein levels in Bregs. The data of boxplots are presented as median (IQR). (ANOVA+Dunnett’s test), compared with group a. Each bubble in boxplots presents data obtained from one sample.