mMIP-1α-DT390核酸制剂治疗实验性变态反应性脑脊髓炎的研究  

Therapeutic effect of a new recombinant immunotoxin mMIP-1α-DT390 on experimental autoimmune encephalomyelitis

作　　者：吕梅励[1] 李虹[1] 梁伟波[1] 陈文捷[1] 贾怡[1] 李明远[1] 蒋忠华[1] 张林[1] 

机构地区：[1]四川大学华西基础医学与法医学院

出　　处：《南方医科大学学报》2007年第6期775-778,共4页Journal of Southern Medical University

基　　金：国家863高科技计划(2002AA214101);国家自然基金委创新研究群体基金(30221001);四川省应用基础项目(04JY029-002)~~

摘　　要：目的新型重组免疫毒素mMIP-1α-DT390核酸制剂对实验性变态反应性脑脊髓炎治疗的效果研究。方法用MBP免疫C57BL/6小鼠诱发EAE,建立人多发性硬化症的动物模型。将构建的真核质粒SRα-mMIP-1α-DT390直接导入EAE模型小鼠C57BL/6肌肉,使重组免疫毒素在其骨骼肌内高效表达。通过对模型小鼠的临床症状的评分、病理切片所显示的中枢神经系统炎症细胞浸润的情况及流式细胞技术检测的外周T细胞的动态变化等指标对核酸制剂的疗效进行评估。结果新型免疫毒素mMIP-1α-DT390核酸制剂治疗EAE模型小鼠,使小鼠临床症状得到改善:发病延迟、平均临床评分下降、症状严重程度降低;神经系统的病理切片显示治疗后小鼠的脑部活化的淋巴细胞浸润减轻;流式细胞仪检测结果显示治疗后小鼠外周血T细胞数量下降,而B淋巴细胞基本维持正常水平。结论所有检测指标均提示该免疫毒素mMIP-1α-DT390核酸制剂可有效的治疗自身免疫性疾病。Objective To evaluate the therapeutic effect of a new recombinant immunotoxin mMIP-1α-DT390 on experimental autoirnmune encephalomyelitis （EAE）. Methods EAE was induced in the low-sensitive strain C57BL/6 mice with intraperitoneal injection of myelin basic protein （MBP） to simulate the human disease multiple sclerosis, followed by intramuscular injection of cationic liposome carrying the plasmid DNA SRα-mMIP-1α-DT390 in the leg muscle to elicit resistance to EAE development. The mice were then examined daily for clinical signs of EAE by an observer blind to the treatment protocol. For immunohistochemistry the mice were anesthetized and perfused with sterile PBS and paraformaldehyde, and the cerebrum, cerebellum, medulla and spinal cord were removed for preparation of serial sections. The mononuclear cells （MNCs） from the EAE mouse spleens were prepared for three-color flow cytometry analysis of the surface markers with appropriate antibodies following the BD Pharmingen cytokine staining protocol. Results EAE model was successfully established by active MBP immunization in C57BL/6 mice. Administration of the immtmotoxin mMIP-1α-DT390 significantly delayed the disease onset and lowered the mean clinical score for EAE as compared with the control mice. Immunohistochemistry demonstrated much less CCR5^＋ infiltrating cells in the central nervous system in mMIP-1α-DT390-treated mice than in the control. The treatment also eliminated reactive T cells in the periphery blood without affecting the number of B cells. Conclusion The immunotoxin mMIP-1α-DT390 can attenuate the disease activity of EAE in mice, suggesting its potential use in the treatment of other autoimmune disorders.

关 键 词：实验性变态性脑脊髓炎 MIP-1Α DT 重组免疫毒素 T细胞 

分 类 号：R744.51[医药卫生—神经病学与精神病学]