Fig. 6

The mechanism underlying the effects of B cells from collagen-induced arthritis (CIA) mice (CIA-B cells) on induced T regulatory cells (iTregs). a, b. CIA-B cells influenced Treg induction in vitro. The percentage of CD4⁺CD25⁺foxp3⁺ cells was detected by flow cytometry after 3 days of co-culture with CIA-B or N-B cells in the presence or absence of transforming growth factor (TGF)-β. c, d CIA-B cells influenced Treg proliferation in vitro. The percentage of carboxyfluorescein succinimidyl ester (CFSE)-labeled iTregs was analyzed by flow cytometry after 3 days of co-culture with CIA-B cells, and some of the medium was treated with the anti-cytotoxic T-lymphocyte-associated protein-4 (anti-CTLA-4) neutralizing antibody or the cells were separated by transwell inserts. e CTLA-4 expression on iTregs after CIA-B cell-driven over-proliferation was determined by flow cytometry. The results represent pooled data from three independent experiments. Bars show the means ± SEM: **P < 0.01 and ***P < 0.001 for comparison of the indicated groups using the unpaired t test. APC antigen-presenting cells, N-B B cells from naïve DBA1/J mice