人型结核分枝杆菌H37Ra在构建实验性自身免疫性重症肌无力大鼠模型中的应用
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摘要
目的探讨人型结核分枝杆菌H37Ra在以鼠源性乙酰胆碱受体α亚基97-116肽段(R97-116)为免疫原构建重症肌无力动物模型实验中的作用。方法以R97-116为免疫原于Lewis大鼠皮下及足垫进行皮下接种,以是否添加H37Ra为条件将20只动物分为实验组和对照组,同时设置空白对照组(10只)。比较各组间行为学、体重及乙酰胆碱受体抗体滴度差异。结果实验组动物局部炎性反应明显,3只动物死亡,6只出现无力症状,1只未出现明显异常表现,6只有无力症状大鼠抗体均呈阳性。对照组大鼠接种处也有局部炎性反应,仅有2只出现轻度无力症状,其中1只抗体可疑阳性,另1只阴性。实验组大鼠体重[(190. 21±8. 35) g]明显低于对照组[(243. 22±7. 98) g]和空白对照组[(251. 22±5. 81) g],差异有统计学意义(P <0. 05),对照组和空白对照组差异无统计学意义。结论 H37Ra为本实验成功的必要试剂。
Objective To investigate the role of mycobacterium tuberculosis H37 Ra in the establishment of myasthenia gravis animal models with rat acetylcholine receptor alpha subunit 97-116 peptide(R97-116) as an immunogen. Methods Subcutaneous inoculation was carried out in the foot pad and skin of Lewis rats with R97-116 as the immunogen. Twenty rats were divided into the experimental group and control group according to whether H37 Ra was added,and a blank control group was set up at the same time. The difference of behavior,body weight and antibody titers of acetylcholine receptors between the three groups was compared. Results In the experimental group,local inflammation was obvious,three rats died,six showed asthenia and positive antibodies,and one showed no obvious abnormal manifestations. In the control group,local inflammatory reactions were also present,and only two showed symptoms of mild weakness,one of which was suspected of having positive antibodies and the other one was negative. The weight of the experimental group [(190. 21 ± 8. 35) g]was significantly lower than that of the control group [(243. 22 ± 7. 98) g]and the blank control group[(251. 22 ± 5. 81) g],and the difference was statistically significant(P < 0. 05),but there was no statistically significant difference between the control group and the blank control group. Conclusions H37 Ra is a necessary reagent for the success of this experiment.
Objective To investigate the role of mycobacterium tuberculosis H37 Ra in the establishment of myasthenia gravis animal models with rat acetylcholine receptor alpha subunit 97-116 peptide(R97-116) as an immunogen. Methods Subcutaneous inoculation was carried out in the foot pad and skin of Lewis rats with R97-116 as the immunogen. Twenty rats were divided into the experimental group and control group according to whether H37 Ra was added,and a blank control group was set up at the same time. The difference of behavior,body weight and antibody titers of acetylcholine receptors between the three groups was compared. Results In the experimental group,local inflammation was obvious,three rats died,six showed asthenia and positive antibodies,and one showed no obvious abnormal manifestations. In the control group,local inflammatory reactions were also present,and only two showed symptoms of mild weakness,one of which was suspected of having positive antibodies and the other one was negative. The weight of the experimental group [(190. 21 ± 8. 35) g]was significantly lower than that of the control group [(243. 22 ± 7. 98) g]and the blank control group[(251. 22 ± 5. 81) g],and the difference was statistically significant(P < 0. 05),but there was no statistically significant difference between the control group and the blank control group. Conclusions H37 Ra is a necessary reagent for the success of this experiment.
引文
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[9]Gooch J W.Antigenic Determinant[M].New York:Springer,2011:874-874.
[10]Lazaridis K,Baltatzidi V,Trakas N,et al.Characterization of a reproducible rat EAMG model induced with various human acetylcholine receptor domains[J].JNeuroimmunol,2017,303:13-21.
[11]Boradia V M,Patil P,Agnihotri A,et al.Mycobacterium tuberculosis H37Ra:a surrogate for the expression of conserved,multimeric proteins of M.tb H37Rv[J].Microb Cell Fact,2016,15(1):140-157.
[12]何宗林,杜先智.活菌H37Ra与灭活菌H37Rv感染小鼠腹腔巨噬细胞的实验研究[J].中国免疫学杂志,2011,27(8):675-680.
[2]Shan W,Li H,Min Z,et al.Curcumin ameliorates experimental autoimmune myasthenia gravis by diverse immune cells[J].Neurosci Lett,2016,626:25-34.
[3]Verma R,Pinto S M,Patil A H,et al.Quantitative Proteomic and phosphoproteomic analysis of H37Ra and H37Rv strains of mycobacterium tuberculosis[J].J Proteome Res,2017,16(4):1632-1645.
[4]Baggi F,Annoni A,Ubiali F,et al.Breakdown of tolerance to a self-peptide of acetylcholine receptor alphasubunit induces experimental myasthenia gravis in rats[J].J Immunol,2004,172(4):2697-2703.
[5]Lennon V A,Lindstrom J M,Seybold M E.Experimental autoimmune myasthenia:a model of myasthenia gravis in rats and guinea pigs[J].J Exp Med,1975,141(6):1365-1375.
[6]Patrick J,Lindstrom J.Autoimmune response to acetylcholine receptor[J].Science,1973,180(4088):871-872.
[7]Zhang Y,Frutiger S,Hughes G J,et al.Identification of T-cell epitopes of autoantigens using recombinant proteins;studies on experimental autoimmune myasthenia gravis[J].Immunology,1990,71(4):538-543.
[8]Wang C C,Zhang M,Li H,et al.Caspase-1 inhibitor regulates humoral responses in experimental autoimmune myasthenia gravis via IL-6-dependent inhibiton of STAT3[J].Neurosci Lett,2017,656:169-176.
[9]Gooch J W.Antigenic Determinant[M].New York:Springer,2011:874-874.
[10]Lazaridis K,Baltatzidi V,Trakas N,et al.Characterization of a reproducible rat EAMG model induced with various human acetylcholine receptor domains[J].JNeuroimmunol,2017,303:13-21.
[11]Boradia V M,Patil P,Agnihotri A,et al.Mycobacterium tuberculosis H37Ra:a surrogate for the expression of conserved,multimeric proteins of M.tb H37Rv[J].Microb Cell Fact,2016,15(1):140-157.
[12]何宗林,杜先智.活菌H37Ra与灭活菌H37Rv感染小鼠腹腔巨噬细胞的实验研究[J].中国免疫学杂志,2011,27(8):675-680.