Cellular and Molecular Study of Blood-Nerve Barrier

免疫球蛋白的穿透和/或激活的肿块细胞向外周神经系统(PNS)实质的迁移是PNS免疫紊乱的最初关键步骤，包括格林-巴利综合征、IgM神经病和慢性炎症性脱髓鞘多神经根神经病。因此，了解神经内膜组织来源的内皮细胞(PnMEC)的细胞特性是很重要的，因为这些细胞构成了血神经屏障(BNB)的主体。为此，我们开发了一种从牛尾分离和培养PnMECs的方法。PnMEC呈鹅卵石状，对第VIII因子/von Willebrand因子(VWF)抗原具有免疫反应性，对Dil-ac-LDL的摄取呈阳性反应。体外培养的PnMEC葡萄糖转运蛋白1(GLUT1)表达迅速下调。除GM3(NeuAc)和GM3(NeuGc)为主要糖鞘糖脂外，PnMECs还包括PNS实质所共有的GM1、GD1a、GD1b和GT1b，以及少量的唾液酸乳糖氨基副糖苷(SLPG)。由于牛PnMECs增殖迅速，可获得大量细胞，该方法可用于PnMECs表面分子的生化分析，这些分子可能在BNB的形成中起关键作用，以及涉及PNS的病理情况。

Penetration of immunoglobulins and/or migration of activated lumphocytes into peripheral nervous system (PNS) parenchyma are the initial key steps to develop immunological disorders of PNS including Guillain-Barre syndrome, IgM neuropathy and chronic inflammatory demyelinating polyradiculoneuropathy. Hence, it is important to know the cellular property of endothelial cells of endoneurial tissue origin (PnMEC) because these cells constitute the bulk of the blood-nerve barrier (BNB). For this purpose we developed a method to isolate and culture pure populations of PnMECs from bovine cauda euina. PnMECs were identified by their cobblestone appearance, immunoreactivity against Factor VIII/von Willebrand factor (vWF) antigen, and positive uptake of Dil-Ac-LDL.The glucose transporter type 1 (GLUT1) expression of these cells was rapidly down-regulated in vitro. Other thatn GM3 (NeuAc) and GM3 (NeuGc) as major glycosphingolipids, PnMECs comprise GM1, GD1a, GD1b and GT1b, which are shared by PNS parenchyma, and sialyl lactosaminyl paragloboside (SLPG) as minor species. Because bovine PnMECs proliferate rapidly and a large mass of cells could be obtained, this method should contribute to the biochemical analysis of surface molecules in PnMECs that might play a key role in the formation of BNB as well as in pathological conditions involving the PNS.