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目的研究B细胞连接蛋白BLNK(B-cell linker protein)的表达对小鼠实验性自身免疫性脑脊髓炎(experimentalautoimmune encephalomyelitis,EAE)的影响并初步探讨其可能的机制。方法 MOG35-55多肽免疫BLNK缺陷(BLNK-deficient,BLNK-/-)鼠及C57BL/6鼠制备EAE小鼠模型,观察实验动物的临床症状及中枢神经系统的病理学变化;应用ELISA方法检测血清中的MOG特异性IgG和IgM抗体水平;采用流式细胞术检测脾脏中调节性T细胞的变化;分选的脾脏B淋巴细胞采用RT-PCR技术评价B细胞所分泌的细胞因子的变化。结果 BLNK-/-鼠临床症状评分明显高于C57BL/6鼠(P<0.01);HE染色及LFB染色结果显示,BLNK-/-鼠与C57BL/C鼠比较炎症感染灶及脱髓鞘病灶明显增多。BLNK-/-鼠血清中的MOG特异性IgG和IgM抗体水平显著低于C57BL/C鼠(P<0.005)。BLNK-/-鼠与C57BL/6鼠相比,脾脏中调节性T细胞百分比及B细胞所分泌的IL-10的表达明显减少,而IL-2的表达显著增高(P<0.05);IFN-γ的表达水平在2组小鼠之间无统计学差异(P>0.05)。结论 BLNK的表达抑制EAE,其机制可能是通过对B细胞分泌的Th1/Th2细胞因子的调节来实现。
Objective To investigate the effect of B-cell linker protein (B-cell linker protein) on experimental autoimmune encephalomyelitis (EAE) in mice and to explore its possible mechanism. Methods The mice model of EAE was induced by BL21 (BLNK-deficient) BL21 (BLNK-deficient) and C57BL / 6 mice by MOG35-55 peptide. The clinical symptoms and pathological changes of central nervous system were observed. MOG specific IgG and IgM antibody levels were measured. The changes of regulatory T cells in the spleen were detected by flow cytometry. The sorted B lymphocytes were used to evaluate the changes of cytokines secreted by B cells by RT-PCR. Results The clinical symptom scores of BLNK - / - mice were significantly higher than those of C57BL / 6 mice (P <0.01). The results of HE staining and LFB staining showed that the inflammatory lesions and demyelinating lesions were significantly different between BLNK - / - mice and C57BL / C mice increase. The levels of MOG-specific IgG and IgM antibodies in BLNK - / - mouse serum were significantly lower than those in C57BL / C mice (P <0.005). Compared with C57BL / 6 mice, the percentage of regulatory T cells in spleen and IL-10 secreted by B cells in BLNK - / - mice was significantly decreased, while the expression of IL-2 was significantly increased (P <0.05) The expression level of γ was not significantly different between the two groups (P> 0.05). Conclusion The expression of BLNK inhibits EAE. The mechanism may be through the regulation of Th1 / Th2 cytokines secreted by B cells.