Taylor D W, Butterworth A E
Parasitology. 1982 Feb;84(1):65-82. doi: 10.1017/s0031182000051672.
Monoclonal antibodies have been produced after fusion of NS-1 murine myeloma cells with spleen cells from mice immunized either by chronic primary infection or with irradiated cercariae: in both cases, animals were challenged with liver cercariae 7 days before fusion. The initial cultures were screened for anti-schistosomular antibodies both by a radioimmunoassay with whole schistosomulum extracts and by immunofluorescence. There was no correlation between the two techniques and subsequent screening was carried out by immunofluorescence. Cloning was carried out in soft agar and 7 cloned cell lines, from 5 initial cultures, were selected for detailed study. Products of 6 of these 7 lines were monoclonal, as judged by isoelectricfocusing of [35S]methionine-labelled supernatant fluids, and their binding to live schistosomula was specific. None of the antibodies showed detectable activity in mediating eosinophil- or complement-dependent damage to schistosomula in vitro. However, 2 antibodies were successfully used to isolate surface proteins with an apparent molecular weight of 24 000 on SDS-polyacrylamide gel electrophoresis.
将NS-1小鼠骨髓瘤细胞与经慢性初次感染或经辐照尾蚴免疫的小鼠的脾细胞融合后,产生了单克隆抗体:在这两种情况下,动物在融合前7天用肝期尾蚴进行攻击。最初的培养物通过用全血吸虫提取物的放射免疫测定法和免疫荧光法筛选抗血吸虫抗体。这两种技术之间没有相关性,随后通过免疫荧光法进行筛选。在软琼脂中进行克隆,并从5个初始培养物中选择7个克隆细胞系进行详细研究。根据[35S]甲硫氨酸标记的上清液的等电聚焦判断,这7个细胞系中的6个细胞系的产物是单克隆的,并且它们与活血吸虫的结合是特异性的。在体外介导嗜酸性粒细胞或补体依赖性对血吸虫的损伤方面,没有一种抗体显示出可检测的活性。然而,两种抗体成功地用于在SDS-聚丙烯酰胺凝胶电泳上分离出表观分子量为24000的表面蛋白。