多发性硬化（MS）是一种CD4+T细胞介导的以中枢脱髓鞘为特征的自身免疫性疾病，神经干细胞（NSCs）可以通过调控CD4+T细胞的增殖分化及细胞因子的分泌治疗MS及其动物模型EAE。B10细胞是一种通过分泌IL-10发挥负性免疫调控作用的Breg细胞。我们在前期研究中发现NSCs治疗组EAE的脾细胞分泌IL-10的能力显著上升，但可以产生IL-10的Treg及Th2细胞比例无显著改变，我们推测B10细胞可能参与其中。为此我们开展预实验发现NSCs治疗组EAE脾脏的B10细胞比例显著上升。在此基础上，本课题通过选择性耗竭或注射外源性B10细胞，明确其是否参与NSCs对EAE的治疗并研究其在NSCs调控免疫中的作用；通过检测B10细胞和CD4+T细胞表面抗原受体及共刺激信号分子的表达、细胞因子的分泌、NSCs的增殖分化及神经营养因子的分泌等，探讨B10细胞参与NSCs调控EAE免疫反应的机制。

Multiple sclerosis (MS) is a CD4+T cell mediated autoimmune demyelinating disorder that target white matter in the central nervous system (CNS). Neural stem cells suppress EAE, the animal model of MS through regulating the proliferation and differentiation of CD4+T cells. B10 cells, one subtype of regulatory B cells, exert immunosuppressive effects by IL-10. In our previous study, we found that splenocytes of EAE mice injected i.v. with NSCs produced higher level of IL-10. However, the proportions of IL-10 producing Treg or Th2 cells did not increase. We hypothesized that IL-10 producing B10 cells may be involved in the regulatory effects of NSCs on EAE splenocytes. Then, result of our pre-experiment showed NSCs increased the proportion of B10 cells in EAE spleen. Based on this finding, we plan to use B10 depleted C57BL/6 mice to study the role of B10 cells in the NSCs therapy and the effects of B10 cells on CD4+T cells. By analyzing the expression of BCR, TCR or co-stimulatory molecules on B10/CD4+T cells, NSC's proliferating/differentiating potential and NSC's ability to secrete neurotrophic factors, we investigate the underlying mechanism.