Koral Gizem, Yılmaz Vuslat, Ulusoy Canan Aysel, Lazaridis Konstantinos, Tüzün Erdem
Department of Neuroscience, Aziz Sancar Institute of Experimental Medicine, Istanbul University;
Department of Neuroscience, Aziz Sancar Institute of Experimental Medicine, Istanbul University.
J Vis Exp. 2025 Jul 3(221). doi: 10.3791/67408.
Myasthenia gravis (MG) is an autoimmune disorder affecting the neuromuscular junction. While most MG patients produce autoantibodies against the acetylcholine receptor (AChR), a subset of patients exhibits autoantibodies targeting the muscle-specific kinase (MuSK). MuSK MG is characterized by severe muscle weakness, treatment-resistant clinical manifestations, and myasthenic crises, often necessitating mechanical ventilation. Consequently, patients frequently require prolonged use of immunosuppressants, which are associated with long-term adverse effects. Given the severity and rarity of MuSK-MG, developing an experimental animal model is crucial for advancing new treatment modalities and a deeper understanding of the underlying pathophysiological mechanisms. Experimental autoimmune myasthenia gravis (EAMG) serves as an animal model for MuSK-MG, effectively but partly mimicking the clinical and immunological features of human MG. The induction of autoimmune animal models can be achieved through active or passive immunization. In the herein presented experimental protocol, active immunization was employed by subcutaneously administering the purified extracellular domain of human MuSK emulsified in complete Freund's adjuvant with heat-killed Mycobacterium tuberculosis. Immunization was performed at four sites, followed by a booster injection of MuSK on the 28th day. This adjuvant with M. tuberculosis enables activation of the immune system through TLR4 and enhanced phagocytosis of the administered antigen. This article comprehensively details the development and characterization of MuSK-EAMG from inception to conclusion.
重症肌无力（MG）是一种影响神经肌肉接头的自身免疫性疾病。虽然大多数MG患者会产生针对乙酰胆碱受体（AChR）的自身抗体，但一部分患者会出现靶向肌肉特异性激酶（MuSK）的自身抗体。MuSK型MG的特征是严重的肌肉无力、难治性临床表现和肌无力危象，常常需要机械通气。因此，患者经常需要长期使用免疫抑制剂，而这些药物会带来长期的不良反应。鉴于MuSK-MG的严重性和罕见性，开发实验动物模型对于推进新的治疗方法以及更深入地了解潜在的病理生理机制至关重要。实验性自身免疫性重症肌无力（EAMG）作为MuSK-MG的动物模型，有效但部分地模拟了人类MG的临床和免疫学特征。自身免疫动物模型的诱导可以通过主动或被动免疫来实现。在本文介绍的实验方案中，主动免疫是通过皮下注射纯化的人MuSK细胞外结构域来实现的，该结构域与热灭活的结核分枝杆菌一起乳化在完全弗氏佐剂中。在四个部位进行免疫，然后在第28天进行MuSK的加强注射。这种含有结核分枝杆菌的佐剂能够通过TLR4激活免疫系统并增强所施用抗原的吞噬作用。本文全面详细地阐述了从开始到结束MuSK-EAMG的建立和特征。
