INTERACTION BETWEEN CEREBRAL CAPILLARY ENDOTHELIUM AND IMMUNE CELLS

最近，我们发现， 从SJL获得的微血管内皮细胞（EC） 与从正常脑中分离的EC相比，患有EAE的小鼠， I-A+，可作为抗原呈递细胞发挥功能。 但 Ia在正常EC表面的表达可被以下物质诱导： γ-干扰素（IFN γ）和随后的EC是 能够呈递抗原（J Immunol 1985; 134：3100和 1986; 137：3428）。 这项研究的中心是发展和 SJL小鼠微血管内皮细胞培养物的建立 易患EAE。 分离内皮细胞 通过匀浆，离心， 不连续蔗糖梯度（1.0-1.5 M）和暴露于分散酶 (0.3%). EC的生长和繁殖依赖于 纤维连接蛋白存在于塑料烧瓶的表面上， 培养基的内皮细胞生长补充剂（GIBCO 199） 含胎牛血清（20%）。 每代样本 的细胞（1-3）用因子VIII抗体染色呈阳性 抗原并且不能用抗神经胶质纤维酸性蛋白染色， 抗巨噬细胞和抗IA。 然而，IA+的表达 抗原和这些细胞的增殖反应是由 将细胞暴露于IFN γ。 这些发现表明 所建立的鼠EC培养物适合于设计 模型系统，以研究大脑 EC和免疫细胞处于活的状态，这在 了解几种神经系统疾病的发病机制 紊乱

Recently, we have shown that freshly isolated cerebromicrovascular endothelial cells (EC) obtained from SJL mice with EAE in contrast to EC separated from normal brain are I-A+ and can function as antigen presenting cells. However, the expression of Ia on the surface of normal EC can be induced by gamma-interferon, (IFN gamma) and subsequently the EC are capable of presenting antigen (J Immunol 1985; 134:3100 and 1986; 137:3428). This study centered in developing and establishing EC cultures derived from microvessels of SJL mice susceptible to EAE. The dissociated endothelium was obtained from cerebral microvessels by homogenization, centrifugation, discontinous sucrose gradient (1.0-1.5 M) and exposure to dispase (0.3%). The growth and propagation of the EC depended on fibronectin presence on the surface of plastic flasks and on endothelial cell growth supplement of the medium (GIBCO 199) containing fetal calf serum (20%). Samples from each generation of cells (1-3) stained positively with antibody to factor VIII antigen and failed to stain with anti-glial fibrilary acidic protein, anti-macrophage and anti-IA. However the expression of IA+ antigen and proliferative response of these cells was induced by exposure of the cells to IFN gamma. These findings indicate that the established murine EC culture is suitable for designing a model system to investigate the interaction between the cerebral EC and immune cells in living state, which are of importance in the understanding of the pathogenesis of several neurological disorders.