骨髓间充质干细胞诱导同种异体B细胞向CD5^+调节性B细胞转化体外实验

Allograft bone marrow mesenchymal stem cells induced B lymphocytes to differentiate into CD5^+ regulatory B cells in vitro

目的研究骨髓间充质干细胞(BMSC)诱导高分泌IL-10的调节性B细胞(Breg)的产生及机制。方法采用成功分离的DA大鼠BMSC与Wistar大鼠B细胞共培养(BMSC+B)96h后,ELISA检测BMSC+B组细胞培养上清中IL-10的浓度和分泌细胞因子的变化,设单纯BMSC、单纯B细胞两组对照;流式细胞术分析B细胞表型的变化;通过加入中和抗体,观察细胞因子对BMSC诱导Breg细胞产生的影响。结果 BMSC+B组成功诱导产生高分泌IL-10的Breg细胞(P=0.006);Breg表型为CD19+CD1d+CD5+;IFN-β促进BMSC诱导Breg的产生(P=0.034)。结论 BMSC能诱导高分泌IL-10的CD5+Breg产生,该机制可能与BMSC自身分泌的IFN-β有关。

Objective To investigate the influence of bone marrow mesenchymal stem cells（BMSCs） on inducing regulatory B cells（Bregs） proliferation and the underlying mechanisim.Methods DA rat BMSCs were cocultured with Wistar rat B cells（BMSCs＋B） for 96 h.Levels of IL-10 and proinflammatory cytokines in supernatants were determined by ELISA and BMSCs and B cells alone groups served as controls.The B-cell phenotype was also analyzed by flow cytometry both in freshly isolated B-cell populations and in B cells that had been cultured with BMSCs.Specific neutralizing monoclonal antibodies were used to test the role of these soluble mediators secreted by BMSCs in inducing IL-10 producing B cells.Results BMSCs＋B group induced B cells to differentiate into Bregs with the highest level of IL-10 after coculture（P=0.006）.These Bregs were with high CD19,CDld,and CD5 expressions.The antibody blockade of IFN-β partially inhibited IL-10 production by B cells induced by BMSCs（P=0.034）.Conclusion BMSCs induced freshly isolated B cells to differentiate into Bregs characterized by the ability to preferentially produce IL-10.IFN-β secreted by BMSCs may partially involve in the production of IL-10 by B cells.