PURIFICATION OF HUMAN EOSINOPHIL-DERIVED NEUROTOXIN
PURIFICATION OF HUMAN EOSINOPHIL-DERIVED NEUROTOXIN
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DOI：
10.1073/pnas.78.8.5165
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
GLEICH, GJ
中科院分区：
文献类型：
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作者：
DURACK, DT;ACKERMAN, SJ;GLEICH, GJ
人和动物嗜酸性粒细胞含有强大的神经毒素，当鞘内注射时，会对实验动物的小脑和脊髓白质造成选择性神经元和轴突损伤。这种反应被称为戈登现象。嗜酸性粒细胞衍生的神经毒素是从 4 名患有各种嗜酸性粒细胞增多综合征的患者的富含嗜酸性粒细胞的白细胞悬浮液或嗜酸性粒细胞颗粒中纯化出来的。通过在 Sephadex G-50 柱上进行连续色谱分离，分离出平均 MW 为 18,400 的单一蛋白质，并通过柱级分的十二烷基硫酸钠/聚丙烯酰胺凝胶电泳进行分析。从这些患者的细胞中纯化的嗜酸性粒细胞衍生的神经毒素在实验动物[兔子]中保留了粗嗜酸性粒细胞或嗜酸性粒细胞颗粒提取物的有效神经毒性活性。这些动物出现僵硬和共济失调综合症，并发展为戈登现象特征的严重瘫痪。对注射了纯化的嗜酸性粒细胞源性神经毒素的动物的大脑进行组织学检查，证实了浦肯野细胞的特征性广泛损失以及小脑、脑干和脊髓白质中严重的海绵状空泡形成。确定了嗜酸性粒细胞颗粒中嗜酸性粒细胞衍生的神经毒素的位置。它与其他几种嗜酸性粒细胞蛋白、颗粒主要碱性蛋白和夏科-莱登晶体蛋白（溶血磷脂酶）不同。
Human and animal eosinophils contain a powerful neurotoxin that causes selective neuronal and axonal damage to white matter of cerebellum and spinal cord of experimental animals when injected intrathecally. This reaction is termed the Gordon phenomenon. The eosinophil-derived neurotoxin was purified from eosinophil-rich leukocyte suspensions, or eosinophil granules from 4 patients with various hypereosinophilic syndromes. A single protein with an average MW of 18,400 was isolated by sequential chromatography on Sephadex G-50 columns and analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis of column fractions. The purified eosinophil-derived neurotoxin from the cells of these patients retained the potent neurotoxic activity of the crude eosinophil or eosinophil granule extracts in experimental animals [rabbits]. The animals developed the syndrome of stiffness and ataxia, progressing to severe paralysis characteristic of the Gordon phenomenon. Histologic examination of the brains of animals injected with purified eosinophil-derived neurotoxin confirmed the characteristic widespread loss of Purkinje cells and severe spongiform vacuolation in the white matter of cerebellum, brain stem and spinal cord. The location of eosinophil-derived neurotoxin in the eosinophil granule was established. It is distinct from several other eosinophil proteins, the granule major basic protein and the Charcot-Leyden crystal protein (lysophospholipase).