MOG-IgG在补体参与下介导脱髓鞘损伤体外模型的建立及应用

The establishment and application of MOG-IgG-mediated complement-dependent in vitro demyelination model

目的通过建立髓鞘少突胶质细胞糖蛋白抗体相关疾病(MOGAD)体外脱髓鞘模型探讨MOGAD的发病机制并筛选疾病治疗药物。方法采用基于转染细胞的亲和层析方法纯化患者来源的髓鞘少突胶质细胞糖蛋白自身抗体(MOG-IgG)。基于大鼠小脑切片和神经元-少突胶质细胞共培养的髓鞘模型,通过MOG-IgG和补体共同作用建立MOGAD体外脱髓鞘模型,将髓鞘碱性蛋白(MBP)的表达量及与神经微丝蛋白(NF)共定位情况作为评估髓鞘形成和损伤的指标。分析多发性硬化促髓鞘再生药物在该模型中的髓鞘修复作用。结果应用转染细胞的亲和层析方法可获得高特异MOG-IgG。经抗体补体作用后,MOG-IgG组MBP表达水平降低(P=0.003),MBP-NF共定位程度降低(P<0.001)。经药物作用后,氯马斯汀组和多潘立酮组MBP表达水平上升(P=0.030,P=0.001),MBP-NF共定位程度增加(P均<0.001)。结论应用该法获得患者来源的高特异MOG-IgG在补体的参与下直接介导髓鞘损伤。氯马斯汀和多潘立酮在MOGAD体外脱髓鞘模型中能促进髓鞘再生。

Objective To explore the pathogenic mechanism of myelin oligodendrocyte glycoprotein-IgG(MOG-IgG)associated disorders(MOGAD)and screen the treatment drugs by establishing the MOGAD in vitro demyelination model.Methods The patient-derived MOG-IgG was purified by using the affinity chromatography based on transfected cells.The MOGAD in vitro demyelination model was constructed by the interaction between MOG-IgG and complement based on rat organotypic cerebellar slice and neuron-oligodendrocyte co-culture model.The expression level of myelin basic protein(MBP)and its colocalization with neurofilament protein(NF)were used as indicators to evaluate the myelin formation and injury.The myelin repair effect of drugs that promote the remyelination for multiple sclerosis was evaluated in this demyelination model.Results Highly-specific MOG-IgG was purified by the affinity chromatography based on transfected cells.After the treatment of antibodies and complement,the expression level of MBP and the colocalization degree of MBP-NF were significantly decreased in the MOG-IgG group(P=0.003,P<0.001).After drug treatment,the expression levels of MBP were significantly up-regulated in the clemastine and domperidone groups(P=0.030,P=0.001),and the colocalization degree of MBP-NF was significantly increased in the clemastine and domperidone groups(both P<0.001).Conclusions Highly-specific patient-derived MOG-IgG obtained by this method directly mediates myelin damage with the participation of complement.Clemastine and domperidone can promote the remyelination in this MOGAD in vitro demyelination model.