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目的:制作不同基因背景小鼠自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)模型,比较不同遗传背景小鼠发病、神经功能评分和病理变化的差异。方法:用髓鞘少突胶质细胞糖蛋白抗原(MOG35-55)免疫C57BL/6和CD-1基因背景小鼠,用完全弗氏佐剂作为抗原载体,并在不同时间点用精制百日咳毒素增强免疫效果,建立自身免疫性脑脊髓炎模型;记录小鼠发病时间与表现,每天进行神经功能评分,并取其脑和脊髓组织进行病理学检查和以CD4、IL-17为靶标的免疫组化染色。结果:C57BL/6组小鼠发病高峰期出现于初次免疫后17～25 d,表现典型的拖尾、单侧或双侧后肢瘫痪等改变,神经功能评分在3分左右;CD-1组小鼠发病高峰期较C57BL/6组推迟,出现于免疫后35～40 d,可见相似的拖尾及偏瘫表现,神经功能评分在2.8分左右。病理检查可见C57BL/6模型小鼠脑、脊髓出现炎症性细胞浸润,而CD-1小鼠的炎性改变相对较轻、且主要出现于脊髓;罗克沙尔固蓝染色法鉴定显示,模型小鼠脑脊髓组织出现脱髓鞘病变,以C57BL/6小鼠更为严重。免疫组织化学法显示2种模型小鼠发病高峰期均存在不同程度的CD4+及IL-17+炎性细胞的浸润。结论:不同的遗传背景对EAE模型发病、临床表现和病理改变有明显影响;CD-1小鼠亦可运用于制作慢性迁延性EAE模型,更符合人类多发性硬化的特点。
OBJECTIVE: To establish a model of experimental autoimmune encephalomyelitis (EAE) in mice with different genetic backgrounds and to compare the differences in the incidence of neurological deficits and pathological changes in mice of different genetic backgrounds. Methods: C57BL / 6 and CD-1 transgenic mice were immunized with myelin oligodendrocyte glycoprotein antigen (MOG35-55), complete Freund’s adjuvant was used as antigen carrier, and at different time points were treated with purified pertussis toxin Enhance the immune effect, establish autoimmune encephalomyelitis model; record the onset time and performance of mice, neurological function score every day, and take the brain and spinal cord tissue pathology and CD4, IL-17-targeted immune group Staining. Results: The peak incidence of C57BL / 6 mice occurred at 17-25 days after primary immunization, showing typical tailing, paralysis of unilateral or bilateral hindlimbs, and neurological function score was about 3 points. CD-1 group was smaller Compared with the C57BL / 6 group, the onset of the mice was postponed 35 to 40 days after immunization, showing similar trailing and hemiparesis performance. The neurological score was about 2.8. Pathological examination showed inflammatory cell infiltration in the brain and spinal cord of C57BL / 6 mice, while inflammatory changes of CD-1 mice were relatively mild, and mainly occurred in the spinal cord. Roxarsal solid-blue staining showed that the model Mice spinal cord tissue demyelinating lesions, C57BL / 6 mice more serious. Immunohistochemistry showed that there were different degrees of infiltration of CD4 + and IL-17 + inflammatory cells in both the model mice at the peak of their onset. CONCLUSION: Different genetic backgrounds have a significant effect on the pathogenesis, clinical manifestations and pathological changes of EAE model. CD-1 mice can also be used for making chronic persistent EAE model and more in line with the characteristics of human multiple sclerosis.