Figure 5.

T follicular helper (Tfh)-like cells help B cells product autoantibody in an IL-21 and CD40 ligand (CD40L)-dependent manner. (A) Tfh-like and B cells from healthy controls (HCs) and experimental autoimmune encephalomyelitis (EAE) mice were cocultured. The level of anti-MOG_35–55 antibodies were measured by chemiluminescent enzyme-linked immunosorbent assay (CLISA) in the supernatant of Tfh/B coculturing system. H Tfh-like and H B stand for Tfh-like from HCs and B cells from HCs, respectively. E Tfh-like and E B stand for Tfh-like from EAE mice and B cells from EAE mice, respectively. ***P < 0.001 vs. the first lane. (B) Sorted B cells from EAE mice were stimulated with MOG_35–55 and relevant cytokines or inhibitors for 7 days. The levels of anti-MOG_35–55 antibodies in the supernatant were measured by CLISA. *P < 0.05, ***P < 0.001 vs. the first lane. (C) Sorted B cells from EAE mice were subjected to the indicated culture conditions. Cells were stimulated for 3 h and then lysed for western blotting analysis to detect the level of the indicated protein markers except B lymphocyte-induced maturation protein-1 (Blimp-1). For Blimp-1, B cells were cultured for 48 h and then prepared for western blotting analysis. NS, nonspecific band. (D) Statistical data of the relative protein level. Data were shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 vs. the first lane. Results are representative of three independent experiments.