CpG-ODN联合抗CD40抗体活化小鼠调节性B细胞抑制CD4＋T细胞增殖  

Regulatory B cells activated by CpG-ODN combined with anti-CD40 monoclonal antibody inhibit CD4^+ T cell proliferation

作　　者：王铿[1] 陶磊[1] 苏剑冰 张悦阳 邹斌华[1] 王祎媛[1] 李晓娟[1] 

机构地区：[1]广东省新药筛选重点实验室,南方医科大学药学院,广东广州510515

出　　处：《细胞与分子免疫学杂志》2016年第9期1164-1167,1173,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81073119,81373123);广东省自然科学基金(2015A030313287)

摘　　要：目的分析CpG寡脱氧核苷酸(CpG-ODN)联合抗CD40抗体刺激后的调节性B细胞(Breg)对其免疫调节功能的影响。方法用流式细胞分选仪分选小鼠脾脏CD5+CD1 dhighBreg与CD5-CD1 dlowB细胞亚群,CpG-ODN联合抗CD40抗体激活24 h,再与纯化的CD4+T细胞共培养,流式细胞术检测活化后的Breg及对照CD5-CD1 dlowB细胞亚群白细胞介素10(IL-10)的表达,以及共培养抗CD3抗体联合抗CD28抗体激活的CD4+T细胞的增殖与γ干扰素(IFN-γ)的水平。结果 CpG-ODN联合抗CD40抗体激活的CD5+CD1 dhighBreg亚群与CD4+T细胞共培养时能明显抑制活化的CD4+T细胞的增殖,同时IFN-γ的水平降低;而CpG-ODN联合抗CD40抗体刺激后的对照B细胞亚群则不能抑制CD4+T细胞的增殖与IFN-γ分泌。CD5+CD1 dhigh Breg亚群在CpG-ODN联合抗CD40抗体刺激24 h,IL-10的表达水平明显高于CD5-CD1 dlowB细胞亚群。结论体外CpG-ODN联合抗CD40抗体刺激后的CD5+CD1 dhighBreg亚群通过分泌IL-10抑制CD4+T细胞的活化。Objective To observe the immunosuppressive function of regulatory B cells （Bregs） in vitro after activated by CpG oligodeoxynucleotide （CpG-ODN） and anti-CD40 mAb. Methods Mice splenic CD5 ＋ CD1dhighB cells and CD5- CD1dlowB cells were sorted by flow cytometry. These B cells were first stimulated with CpG-ODN combined with anti-CD40 mAb for 24 hours, and then co-cultured with purified CD4＋T cells. The interleukin 10 （IL-10） expression in the activated Bregs and other B cell subset, as well as the proliferation and interferon y （IFN-y） expression in the CD4 ＋ T cells activated by anti-CD3 mAb plus anti-CD28 mAb were determined by flow cytometry. Results CD5＋ CD1dhigh B cells activated by CpG-ODN plus anti-CD40 mAb blocked the up-regulated CD4 ＋T proliferation and significantly reduced the IFN-y level. At the same time, activated CD5- CD1dlowB cells showed no inhibitory effect on CD4＋T cells. Further study revealed that IL-10 expression in the CD5 ＋ CD1dhigh B cells were much higher than that in the CD5- CD1dlow B cells after stimulated with CpG-ODN combined with anti-CD40 mAb for 24 hours. Conclusion CD5 ＋ CD1dhighB cells activated by CpG-ODN combined with anti-CD40 mAb have immune inhibitory effects on CD4 ＋ T cell activation in vitro, which possibly due to IL-10 secretion.

关 键 词：调节性B细胞 CPG寡脱氧核苷酸 抗CD40抗体 免疫抑制 CD4+T细胞 

分 类 号：R392.12[医药卫生—免疫学]