Interleukin-10 production and T cell-suppressive capacity in B cell subsets from atherosclerotic apoE ^-/- mice

The evidence regarding the role of regulatory B cells (Breg) in atherosclerosis are scarce, and there are contradictory data about their atheroprotective properties. Due to the demonstrated protective function of Breg in different inflammatory diseases mainly through interleukin-10 (IL-10) production, the knowledge of their participation in atherosclerosis immunopathology would be very valuable. To further study which B cell subsets participate in IL-10 production and their regulatory role, splenocytes from apolipoprotein-E-deficient mice were evaluated by ex vivo and in vitro cultures. Atherosclerotic mice had increased frequency of IL-10⁺ B cells, which presented high CD1d, CD19, and IgM, but variable CD5, CD21, and CD23 expression. IL-10⁺ B cells were not enriched in B cell subsets previously reported as Breg. Increased frequency of IL-10⁺ B cells with transitional 1-like (T1-like) and follicular (FO) and reduced CD5⁺ and marginal zone (MZ) phenotypes were observed ex vivo. Increased frequency of IL-10⁺ B cells with T1-like and MZ, and decreased IL-10⁺ FO and T2 phenotypes were also observed in vitro. To determine regulatory capacity of B cells in the atherosclerotic model, each subset were co-cultured with CD4⁺CD25^- T cells. CD5⁺, FO, MZ, and T1-like cells from atherosclerotic mice exhibited regulation in an IL-10-dependent manner. However, only FO cells decreased both frequency of interferon gamma (IFN-γ)⁺ and tumor necrosis factor alpha (TNF-α)⁺ and proliferation of T cells. Finally, splenocytes showed increased frequency of IFN-γ⁺ and TNF-α⁺ cells only when FO-depleted B cells were evaluated. These results suggest that mainly FO B cells can modulate in some level the inflammatory responses observed in atherosclerosis.