Wang Keng, Tao Lei, Su Jianbing, Zhang Yueyang, Zou Binhua, Wang Yiyuan, Li Xiaojuan
School of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Screening, Southern Medical University, Guangzhou 510515, China.
School of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Screening, Southern Medical University, Guangzhou 510515, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Sep;32(9):1164-7.
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(+)CD1d(high)B cells and CD5(-)CD1d(low)B 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 γ (IFN-γ) expression in the CD4(+) T cells activated by anti-CD3 mAb plus anti-CD28 mAb were determined by flow cytometry. Results CD5(+)CD1d(high) B cells activated by CpG-ODN plus anti-CD40 mAb blocked the up-regulated CD4(+)T proliferation and significantly reduced the IFN-γ level. At the same time, activated CD5(-)CD1d(low)B cells showed no inhibitory effect on CD4(+)T cells. Further study revealed that IL-10 expression in the CD5(+)CD1d(high) B cells were much higher than that in the CD5(-)CD1d(low)B cells after stimulated with CpG-ODN combined with anti-CD40 mAb for 24 hours. Conclusion CD5(+)CD1d(high) B 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.
目的 观察经CpG寡脱氧核苷酸(CpG-ODN)和抗CD40单克隆抗体激活后的调节性B细胞(Bregs)在体外的免疫抑制功能。方法 采用流式细胞术分选小鼠脾脏CD5(+)CD1d(high)B细胞和CD5(-)CD1d(low)B细胞。这些B细胞先用CpG-ODN联合抗CD40单克隆抗体刺激24小时,然后与纯化的CD4(+)T细胞共培养。通过流式细胞术检测活化的Bregs和其他B细胞亚群中白细胞介素10(IL-10)的表达,以及抗CD3单克隆抗体加抗CD28单克隆抗体激活的CD4(+)T细胞的增殖和干扰素γ(IFN-γ)表达。结果 CpG-ODN加抗CD40单克隆抗体激活的CD5(+)CD1d(high)B细胞可阻断CD4(+)T细胞的增殖上调,并显著降低IFN-γ水平。同时,活化的CD5(-)CD1d(low)B细胞对CD4(+)T细胞无抑制作用。进一步研究发现,用CpG-ODN联合抗CD40单克隆抗体刺激24小时后,CD5(+)CD1d(high)B细胞中IL-10的表达远高于CD5(-)CD1d(low)B细胞。结论 CpG-ODN联合抗CD40单克隆抗体激活的CD5(+)CD/d(high)B细胞在体外对CD4(+)T细胞的活化具有免疫抑制作用,这可能是由于IL-10的分泌所致。