Schmitt Jürgen, Wuhrer Manfred, Hamburger Joseph, Jourdane Joseph, Ramzy Reda M R, Geyer Rudolf, Ruppel Andreas
Department of Tropical Hygiene and Public Health, University of Heidelberg, Germany.
J Parasitol. 2002 Jun;88(3):505-13. doi: 10.1645/0022-3395(2002)088[0505:SMASHI]2.0.CO;2.
Two carbohydrate epitopes were identified by monoclonal antibodies (KCS and E2) and characterized with respect to their immunoreactivity, monosaccharide structure, and location. Immunofluorescence demonstrated the presence of both epitopes on the surfaces of sporocysts, cercariae, and miracidia of Schistosoma mansoni, Schistosoma haematobium, and Schistosoma japonicum. However, spatial distribution and density of expression varied among species and developmental stages, and neither epitope was detectable on adult worm surfaces. Both glycans were found in the hemolymph of infected, but not uninfected, intermediate snail hosts. The presence of epitopes in hemolymph, as well as in schistosome eggs, is species-specific for KCS, recognizing only S. mansoni, and partly specific for E2, which reacted predominantly with S. haematobium. Immunoaffinity purification of target antigens for KCS and E2 from hemolymph of infected Biomphalaria and Bulinus, respectively, followed by carbohydrate composition analysis revealed a high content of fucose in both glycans. Methylation analysis demonstrated exclusively terminal fucose for the target antigen of KCS and terminal as well as internal fucose for the one of E2. Removal of terminal fucose abolished reactivity with both monoclonal antibodies. Both glycans are different from previously characterized schistosome carbohydrates. Their biological function(s) remain to be defined.
通过单克隆抗体(KCS和E2)鉴定出两种碳水化合物表位,并对其免疫反应性、单糖结构和位置进行了表征。免疫荧光显示,曼氏血吸虫、埃及血吸虫和日本血吸虫的子孢子、尾蚴和毛蚴表面均存在这两种表位。然而,物种和发育阶段之间的空间分布和表达密度各不相同,在成虫表面均未检测到这两种表位。在受感染而非未受感染的中间螺宿主的血淋巴中发现了这两种聚糖。血淋巴以及血吸虫卵中表位的存在对KCS具有物种特异性,仅识别曼氏血吸虫,对E2则部分具有特异性,E2主要与埃及血吸虫发生反应。分别从受感染的双脐螺和水泡螺的血淋巴中免疫亲和纯化KCS和E2的靶抗原,随后进行碳水化合物组成分析,结果显示两种聚糖中岩藻糖含量都很高。甲基化分析表明,KCS的靶抗原仅含末端岩藻糖,而E2的靶抗原既有末端岩藻糖也有内部岩藻糖。去除末端岩藻糖后,两种单克隆抗体均失去反应活性。这两种聚糖均与先前表征的血吸虫碳水化合物不同。它们的生物学功能尚待确定。