Supplementary Material for: Age-dependent Effects of Transgenic 2D2 Mice Used to Induce Passive EAE in C57BL/6 Mice

Introduction: Multiple sclerosis (MS) is a neurodegenerative autoimmune disease which worsens with age. Here we examined the influence of age on passive experimental autoimmune encephalomyelitis (P-EAE), a model to study MS, using young and mature adult 2D2 transgenic donor mice to induce pathology in WT C57BL6/J mice. Methods: Lymphocytes from young adult (i.e., 10-week-old) or mature adult (i.e., 6-month-old) transgenic donor mice were characterized by flow cytometry prior to injection of cultured leukocytes into adult female WT recipient mice, with a special focus on transgenic T cell phenotypes. Results: Our findings show age-dependent changes in memory T cell phenotypes correlated with more severe clinical and histological disease when donor cells originated from young as compared to mature adult mice. Conclusion: Not only do these results demonstrate that the age of the 2D2 transgenic donor mice is critical in establishing P-EAE, but the differential effects might also identify age-dependent factors that contribute to EAE and perhaps MS.

引言：多发性硬化症（Multiple sclerosis, MS）是一种随年龄进展而加重的神经退行性自身免疫疾病。本研究以青年与成熟成年2D2转基因供体小鼠为实验材料，在野生型C57BL6/J小鼠中诱导病理损伤，以此探究年龄对被动型实验性自身免疫性脑脊髓炎（passive experimental autoimmune encephalomyelitis, P-EAE，一种MS研究动物模型）的影响。
方法：在将培养的白细胞注射至成年雌性野生型受体小鼠前，通过流式细胞术（flow cytometry）对青年成年（10周龄）或成熟成年（6月龄）转基因供体小鼠的淋巴细胞进行表型表征，重点关注转基因T细胞表型。
结果：相较于成熟成年供体小鼠，青年供体来源的细胞所诱导的疾病临床症状与组织病理学损伤更为严重，该现象与记忆性T细胞表型的年龄依赖性变化存在相关性。
结论：本研究结果不仅证实2D2转基因供体小鼠的年龄在构建P-EAE模型中至关重要，其诱导的差异效应或可揭示影响EAE乃至多发性硬化症的年龄依赖性致病因子。