Department of Pathology and Lab Medicine, University of Wisconsin-Madison, Madison, Wisconsin.
Department of Microbiology-Immunology, Northwestern University Feinberg School of Medicine, Chicago, Illinois.
Curr Protoc. 2021 Dec;1(12):e300. doi: 10.1002/cpz1.300.
This article details the materials and methods required for both active induction and adoptive transfer of experimental autoimmune encephalomyelitis (EAE) in the SJL mouse strain using intact proteins or peptides from the two major myelin proteins: proteolipid protein (PLP) and myelin basic protein (MBP). Additionally, active induction of EAE in the C57BL/6 strain using myelin oligodendrocyte glycoprotein (MOG) peptide is also discussed. Detailed materials and methods required for the purification of both PLP and MBP are described, and a protocol for isolating CNS-infiltrating lymphocytes in EAE mice is included. Modifications of the specified protocols may be necessary for efficient induction of active or adoptive EAE in other mouse strains. © 2021 Wiley Periodicals LLC. Basic Protocol: Active induction of EAE with PLP, MBP, and MOG protein or peptide Alternate Protocol: Adoptive induction of EAE with PLP-, MBP-, or MOG-specific lymphocytes Support Protocol 1: Purification of proteolipid protein Support Protocol 2: Purification of myelin basic protein Support Protocol 3: Isolation of CNS-infiltrating lymphocytes.
这篇文章详细介绍了使用两种主要髓鞘蛋白(髓鞘少突胶质细胞糖蛋白和髓鞘碱性蛋白)的完整蛋白或肽在 SJL 小鼠中主动诱导和过继转移实验性自身免疫性脑脊髓炎(EAE)所需的材料和方法。此外,还讨论了使用髓鞘少突胶质细胞糖蛋白(MOG)肽在 C57BL/6 品系中主动诱导 EAE 的方法。本文详细描述了 PLP 和 MBP 的纯化所需的材料和方法,并包含了在 EAE 小鼠中分离中枢神经系统浸润淋巴细胞的方案。对于在其他小鼠品系中有效诱导主动或过继性 EAE,可能需要对指定方案进行修改。© 2021 Wiley Periodicals LLC. 基本方案:用 PLP、MBP 和 MOG 蛋白或肽主动诱导 EAE 备选方案:用 PLP、MBP 或 MOG 特异性淋巴细胞过继诱导 EAE 支持方案 1:髓鞘少突胶质细胞糖蛋白的纯化 支持方案 2:髓鞘碱性蛋白的纯化 支持方案 3:中枢神经系统浸润淋巴细胞的分离