多发性硬化症（Multiple sclerosis (MS)）/实验性自身免疫性脑脊髓炎模型（EAE）研究专题

 关键词：多发性硬化症，Multiple sclerosis，EAE，MOG, MOG35-55，PLP，PLP (139-151)，实验性自身免疫性脑脊髓炎

多发性硬化症（Multiple sclerosis (MS)）/实验性自身免疫性脑脊髓炎模型（EAE）研究专题

背景介绍

多发性硬化症(multiple sclerosis, MS)是一种原发于中枢神经系统的炎症性脱髓鞘疾病。MS确切的发病机制虽尚不清楚, 但普遍认为是在易感基因的基础上, 受环境因素触发, 由CD4⁺ T细胞介导的中枢神经系统(central nervous system, CNS)自身免疫性疾病。实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis, EAE)是一种以特异性致敏的CD4⁺ T细胞介导为主, 对实验动物进行髓鞘蛋白免疫构建的疾病模型。EAE与人类MS在临床、生化、免疫及病理等诸多方面具有相同的特征, 所以它是目前国际公认的 MS理想动物模型。

（一）慢性EAE（Chronic EAE）模型的建立

髓鞘少突胶质细胞糖蛋白(MOG) 是髓鞘膜和少突胶质细胞表面最外层的膜蛋白，是导致多发性硬化（MS）脱髓鞘的关键成分，针对MOG的抗体能够在体内和体外造成脱髓鞘。MOG数量较少，约占髓鞘蛋白总量的0.01%-0.05%，具有高度免疫原性，它直接参与CNS的体液免疫反应，是引起实验性自身免疫性脑脊髓炎（EAE）中脱髓鞘的主要免疫靶点。MOG 35-55含21个氨基酸，是唯一既能引起脱髓鞘抗体反应又能引起T细胞反应的中枢神经系统髓鞘蛋白成分。MOG 35-55可在C57BL/6小鼠体内诱导MOG35-55/I-Ab特异性自身反应性CD4⁺T细胞异常活化，诱发EAE。

产品性质

运输与保存方法

冰袋（wet ice）运输。收到产品后放到-20 ºC保存，保持干燥。不建议溶液状态保存。

产品信息

Q&A

1. 如何选择模型动物？

答：一般选择雌性C57BL/6小鼠，10-14周左右。

2.在什么部位注射？每次注射的量？

皮下注射，每只小鼠200μg MOG 35-55多肽。

参考文献

1.Mandy?L. Ford, et al. An MHC anchor-substituted analog of myelin oligodendrocyte glycoprotein?35–55 induces IFN-γ and autoantibodies in the absence of experimental autoimmune encephalomyelitis and optic neuritis. European Journal of Immunology Volume 34, Issue 2, pages 388–397,February 2004

2.Cathleen Rich, et al. Myelin oligodendrocyte glycoprotein-35–55 peptide induces severe chronic experimental autoimmune encephalomyelitis in HLA-DR2-transgenic mice. Eur. J. Immunol. 2004. 34: 1251–1261

3.Cong-Qiu Chu, et al. Failure to Suppress the Expansion of the Activated Cd4 T Cell Population in Interferon γ–Deficient Mice Leads to Exacerbation of Experimental Autoimmune Encephalomyelitis. JEM Home 2000 Archive 3 July Chu et al. 192 (1): 123.

4. Stefanie Kuerten, et al. MP4- and MOG:35–55-induced EAE in C57BL/6 mice differentially targets brain, spinal cord and cerebellum. Journal of Neuroimmunology Volume 189, Issues 1–2, 2007 (31–40).

5. Anthony Slavin, et al. Induction of a Multiple Sclerosis-Like Disease in Mice with an Immunodominant Epitope of Myelin Oligodendrocyte Glycoprotein. 1998, Vol. 28, No. 2 , 109-120.

6. McFarland, H. F., et al. Multiple sclerosis: a complicated picture of autoimmunity. Nat. Immunol. 8, 913-919, doi:10.1038/ni1507

(2007).

7. Wiendl, H. Neuroinflammation: the world is not enough. Curr. Opin. Neurol. 25, 302-305, doi:10.1097/WCO.0b013e3283534abf (2012).

8. van der Star, B. J. et al. In vitro and in vivo models of multiple sclerosis. CNS Neurol. Disord. Drug Targets. 11, 570-588 (2012).

9. Pachner, A. R. Experimental models of multiple sclerosis. Curr. Opin. Neurol. 24, 291-299, doi:10.1097/WCO.0b013e328346c226 (2011).

10. Bittner, S. et al. The TASK1 channel inhibitor A293 shows efficacy in a mouse model of multiple sclerosis. Exp. Neurol. 238, 149-155, doi:10.1016/j.expneurol.2012.08.021 (2012).

（二）复发-缓解型EAE（Relapsing- Remitting EAE）模型的建立

复发-缓解型MS是人类多发性硬化（MS）最常见的一种，这类型的疾病与PLP_139-151/CFA诱导的SJL小鼠EAE症状最为相似，因此，这是一种常用于研究药物对复发型EAE作用的最好模型。而且，该模型也表现出较高的EAE同步发病率。复发-缓解型EAE常用于MS治疗，以研究EAE复发的进展情况。Proteolipid Protein (PLP)是一种高度疏水的膜蛋白，是中枢神经系统(central nervous system, CNS)中含量最多的髓磷脂。与IA^S MHCII结合时，PLP 139–151和PLP 178–191具有相同的高亲和力，但是，免疫系统对PLP 139–151表位产生的免疫反应是主要的。对于PLP诱导的EAE模型，SJL (H-2^s)小鼠具有很高的致敏性。

产品性质

运输与保存方法

冰袋（wet ice）运输。收到产品后放到-20 ºC保存，保持干燥。不建议溶液状态保存。

产品信息

Direct EAE induction in SJL mice

Q&A

1. 如何选择模型动物？

答：一般选择雌性SJL小鼠，9-10周左右。

2.在什么部位注射？每次注射的量？

皮下注射，每只小鼠100μg PLP_139-151 。

参考文献

1. McRae BL, et al. Differential recognition of peptide analogs by naive verses activated PLP 139-151-specific CD4+ T cells. J Neuroimmunol. 1995 Jul;60(1-2):17-28.

2. Ana C. Anderson, et al. High Frequency of Autoreactive Myelin Proteolipid Protein–Specific T Cells in the Periphery of Naive Mice. JEM vol. 191 (5): 761-770.

3. Maria Katsara, et al. Immune responses of linear and cyclic PLP139-151 mutant peptides in SJL/J mice: peptides in their free state versus mannan conjugation. Immunotherapy, 6(6): 709-724. DOI 10.2217/imt.14.42

4. Stephen D. Miller, et al. Experimental Autoimmune Encephalomyelitis in the Mouse. Curr Protoc Immunol. 2007 May ; CHAPTER: Unit–15.1. doi:10.1002/0471142735.im1501s77.

5. Hanspeter Waldner, et al. Fulminant spontaneous autoimmunity of the central nervous system in mice transgenic for the myelin proteolipid protein-specific T cell receptor. PNAS March 28, 2000 97(7) 3412–3417.

6. By Ana C. Anderson, et al. High Frequency of Autoreactive Myelin Proteolipid Protein–specific T Cells in the Periphery of Naive Mice: Mechanisms of Selection of the Self-reactive Repertoire. J. Exp. Med. 191(5) 2000: 761–770.

7. Harald H. Hofstetter, et al. Pertussis Toxin Modulates the Immune Response to Neuroantigens Injected in Incomplete Freund’s Adjuvant: Induction of Th1 Cells and Experimental Autoimmune Encephalomyelitis in the Presence of High Frequencies of Th2 Cells. J Immunol 2002; 169:117-125.