Immunotherapy against amyloid beta is modulated by meningeal lymphatic function I. Immunotherapy against amyloid beta is modulated by meningeal lymphatic function I

A growing body of evidence points to passive immunotherapy as a promising therapeutic strategy against AD. Herein, we show that meningeal lymphatic vasculature becomes dysfunctional in AD transgenic mice and that manipulating meningeal lymphatic drainage affects the outcome of anti-Aβ immunotherapy. Overall design: Lymphatic endothelial cells (LECs) were isolated from the meninges of male 5xFAD mice, or non-transgenic wild type age-matched littermates (WT), at the age of 6 months. LECs were sorted by FACS directly into lysis buffer according to the following phenotype: Singlets DAPI-CD45-CD31+PDPN+. Each individual sample results from meningeal cells pooled from 10 mice.

越来越多的证据表明，被动免疫疗法（passive immunotherapy）是对抗阿尔茨海默病（AD, Alzheimer's Disease）的极具前景的治疗策略之一。本研究证实，AD转基因小鼠的脑膜淋巴血管系统会出现功能障碍，且调控脑膜淋巴引流可影响抗β淀粉样蛋白（Aβ）免疫疗法的治疗效果。
实验整体设计如下：我们于小鼠6月龄时，从雄性5xFAD AD转基因小鼠以及同月龄的非转基因野生型（WT, wild type）同窝对照小鼠的脑膜组织中分离淋巴管内皮细胞（LECs, lymphatic endothelial cells）。随后依据以下细胞表型，通过荧光激活细胞分选术（FACS, Fluorescence-Activated Cell Sorting）将LECs直接分选至裂解液中：单重体细胞（Singlets）、DAPI⁻CD45⁻CD31⁺PDPN⁺。每份独立样本均由10只小鼠的脑膜细胞混合制备得到。