Characterization of a stage-specific Mr16 000 schistosomular surface glycoprotein antigen of Schistosoma mansoni
Characterization of a stage-specific Mr16 000 schistosomular surface glycoprotein antigen of Schistosoma mansoni
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DOI：
10.1016/s0166-6851(99)00041-9
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发表时间：
1999-05-15
影响因子：
1.5
通讯作者：
Oldridge, J
中科院分区：
文献类型：
--
作者：
Bickle, QD;Oldridge, J
16 kDa曼氏血吸虫血吸虫表面抗原(Sm16)最初被描述为被动保护性单抗(B3A)的靶标。它出现在尾虫转化后的血吸虫表面，并被暴露于减毒尾虫的动物的血清唯一识别。在这项工作中，用Triton X-114和十二烷基硫酸钠对血吸虫幼虫的连续提取表明，Sm16是一个完整的膜结构，而不是由磷脂酰肌醇特异性磷脂酶C实验判断的糖基磷脂酰肌醇锚定。Sm16在感染曼氏血吸虫的钉螺的肝胰腺中表达，在尾蚴和机械转化的血吸虫中同样丰富，但在肝期虫卵中未检测到Sm16。免疫电子显微镜显示Sm16在尾蚴中定位于被认为是充满膜性小泡的被下细胞体。用蛋白酶和偏碘酸钠处理表明Sm16是一种糖蛋白，B3A识别的表位是高碘酸盐敏感的。双向电泳法测得等电点为6。N-糖苷酶F、内切糖苷酶F或内切-α-N-乙酰半乳糖胺酶对B3A的大小和识别均无影响。用广泛的生物素化凝集素进行的免疫印迹显示，只有花生凝集素和蓖麻凝集素II(蓖麻毒素)才能识别。IR结果表明，Sm16具有抗原表面暴露的O-连接复合寡糖，不含甘露糖/葡萄糖、GlcNAc、L岩藻糖和唾液酸，但含有末端Galβ(1-3)GalNAc和/或半乳糖。(C)1999 Elsevier Science B.V.保留所有权利。
A 16 kDa Schistosoma mansoni schistosomular surface antigen (Sm16) was originally described as the target of a passively protective mAb (B3A). It appeared on the schistosomular surface after transformation of cercariae and was uniquely recognised by sera from animals exposed to attenuated cercariae. In this work sequential extractions of schistosomula with Triton X-114 and sodium dodecyl sulphate showed Sm16 to be an integral membrane structure which did not appear to be glycosylphosphatidylinositol-anchored as judged by experiments using phosphatidyl inositol-specific phospholipase C. The antigen was strongly reactive in Western blotting with rabbit irradiated vaccine sera. Sm16 was demonstrated in the hepatopancreas of S. mansoni-infected snails and was equally abundant in cercariae and mechanically- transformed schistosomula but was undetected in liver stage worms or eggs. Immunoelectron microscopy showed Sm16 to be localised, in cercariae, to what are believed to be subtegumental cell bodies packed with membraneous vesicles. Treatment with proteases and with sodium metaperiodate showed Sm16 to be a glycoprotein of which the epitope recognised by B3A was periodate sensitive. Two-dimensional electrophoresis gave a PI of 6. Neither the size or the recognition by B3A was affected by treatment with N-glycosidase F, endoglycosidase F or endo-alpha-N-acetylgalactosaminidase. Western blotting using a wide range of biotinylated lectins showed recognition only by peanut agglutinin and Ricinus communis agglutinin II (ricin). Ir is concluded that Sm16 has antigenic surface-exposed O-linked complex oligosaccharides which lack mannose/glucose, GlcNAc, L-fucose and sialic acid but contain terminal Gal beta (1-3) GalNAc and/or galactose. (C) 1999 Elsevier Science B.V. All rights reserved.