目的 探索利用基因工程生产的少突胶质细胞糖蛋白-硫氧还融合蛋白(MOG-TrxA)免疫C57BL/6(H-2b)小鼠建立实验性自身免疫性脑脊髓炎(EAE)动物模型的方法.方法 利用实验室已经重组成功的携带pET32(a)-MOG-TrxA质粒的大肠杆菌BL21和携带pET32(a)-TrxA质粒的空载大肠杆菌BL21分别诱导大量表达MOG-TrxA和TrxA,并利用蛋白金属螯合亲和层析纯化,超滤法浓缩除盐,Bradford法蛋白定量,SDS聚丙烯酰胺凝胶电泳鉴定.利用制备的MOG-TrxA抗原免疫C57BL/6(H-2b)小鼠诱导EAE动物模型,以TrxA免疫动物作为对照组,以脊髓匀浆(SCH)免疫动物作为阳性对照组,以临床评分、体重变化、HE染色和Klüver-Barrera髓鞘染色评分等指标,评价模型的质量.结果 MOG-TrxA组动物在临床评分、HE染色病理评分和Klüver-Barrera髓鞘染色病理评分等指标与TrxA组相比明显增高,体重明显下降,数据差异均有统计学意义(均P＜0.01),与SCH组相比差异无统计学意义(P＞0.05).结论 利用基因工程制备的MOG-TrxA免疫C57BL/6(H-2b)小鼠能成功建立慢性非缓解型EAE动物模型.%Objective To establish mouse experimental autoimmune encephalomyelitis (EAE) model induced by MOG-TrxA immunization of C57BL/6(H-2b) mice engineering. Methods MOG-TrxA and TrxA was expressed in E. coli BL21 by transfection of recombinant plasmids pET32(a)-MOG-TrxA and pET32(a)-TrxA respectively, The purified proteins were applied to immunize C57BL/6 (H-2b) mice to induce EAE (MOG-TrxA) and control (TrxA). The mice immunized with spinal cord homogenate (SCH) as the positive control. The clinical scores curve, weight variation curve, HE staining and Klü ver-Barrera staining scores were observed to evaluate quality of EAE model. Results There were significant differences in clinical scores, weight changes, HE staining scores and Klüver-Barrera staining scores between MOG-TrxA group and TrxA group (P ＜0.05), while there were no significant differences between MOG-TrxA group and SCH group (P ＞0.05). Conclusion The EAE animal model has been successful established by immunization of gene-engineered MOG-TrxA in C57BL/6(H-2b) mice.
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