IL-35改善肌肉特异性酪氨酸激酶重症肌无力小鼠症状  

IL-35 ameliorates muscle weakness in MuSK-immunized mice

作　　者：刘潺潺 李婷 徐安定[1] 吴本清 LIU Chan-chan;LI Ting;XU An-ding;WU Ben-qing(Clinical Neuroscience Institute,Stroke Center&Department of Neurology,The First Affiliated Hospital,Jinan University,Guangzhou 510632,China;University of Chinese Academy of Sciences-Shenzhen Hospital,Shenzhen 518000,China)

机构地区：[1]暨南大学附属第一医院临床神经科学研究所,广东广州510632 [2]中国科学院大学深圳医院(光明),广东深圳518000

出　　处：《生物技术》2021年第6期601-606,共6页Biotechnology

基　　金：深圳市光明区经济发展专项资金资助(2020R01001);中国博士后科学基金新冠肺炎疫情防控专项资助(2020T130047ZX);深圳市卫计委研究项目(SZBC2017018)。

摘　　要：[目的]探索IL-35是否可以缓解肌肉特异性酪氨酸激酶(MuSK)抗体阳性的实验室自身免疫性重症肌无力模型(EAMG)小鼠的症状。[方法]使用IL-35慢病毒载体感染外周血单核细胞(PBMC),慢病毒pLVX-IL35的效价为:6×10^(7) TU/mL。将小鼠随机分为3组:(1)模型小鼠+PBMCs为对照组;(2)模型小鼠+空病毒转染的PBMCs;(3)模型小鼠+转染IL-35慢病毒的PBMCs。将转染的PBMC静脉注射到小鼠模型中,在第二次注射PBMC之后的第7 d,每天评估临床评分。采用ELISA检测血清IL-2、IL-10和TGF-β水平。RT-PCR检测小鼠血清中的TGF-β、EBI3、IL-12A和IL-10。流式细胞仪检测外周血细胞中CD4^(+)CD25^(+)Treg的百分比变化。每周一次翻笼试验。[结果]相比对照组,静脉注射IL-35慢病毒载体的MuSK免疫的EAMG小鼠,能够表达IL-35(P<0.01),降低小鼠的临床评分(P<0.05),增加IL-10(P<0.01)和TGF-β1(P<0.01)。[结论]IL-35可以优化12w后MuSK抗体阳性重症肌无力(MG)小鼠的临床评分(P<0.01)和翻笼试验时间(P<0.01),能成为治疗作用靶点。[Objective]To explore whether IL-35 can alleviate the symptoms of laboratory autoimmune myasthenia gravis model(EAMG)with positive muscle-specific tyrosine kinase(MuSK)antibodies.[Method]The IL-35 lentiviral vector was used to infect peripheral blood mononuclear cells(PBMC).The titer of the lentivirus pLVX-IL35 was 6×10^(7) TU/mL.The mice were randomly divided into 3 groups:(1)model mice+PBMCs as the control group(2)model mice+PBMCs transfected with empty virus(3)model mice+PBMCs transfected with IL-35 lentivirus.The transfected PBMC was injected intravenously into the mouse model,and the clinical score was evaluated daily on the 7 th day after the second injection of PBMC.ELISA was used to detect serum IL-2,IL-10,TGF-βand antibody titers.RT-PCR detects TGF-β,EBI3,IL-12 A,IL-10 in mouse serum.Flow cytometry was used to detect the percentage change of CD4^(+)CD25^(+)Treg in peripheral blood cells.Cage turning test once a week.[Result]IL-35 lentiviral vector was injected intravenously with MuSK-immunized EAMG,which was able to express IL-35(P<0.01),reduced EAMG clinical score(P<0.05),IL-10(P<0.01)and TGF-β1(P<0.01)increase.[Conclusion]IL-35 can optimize the clinical grades(P<0.01)and the inverted screen of the mice(P<0.01)of MuSK antibody-positive myasthenia gravis(MG)after 12 weeks,and can become a therapeutic target.

关 键 词：重症肌无力 实验性自身免疫性重症肌无力模型(EAMG) 肌肉特异性激酶 IL-35 细胞因子 

分 类 号：Q956[生物学—动物学]