Robijn M L M, Wuhrer M, Kornelis D, Deelder A M, Geyer R, Hokke C H
Department of Parasitology, Centre of Infectious Diseases, Leiden University Medical Centre, 2300 RC Leiden, The Netherlands.
Parasitology. 2005 Jan;130(Pt 1):67-77. doi: 10.1017/s0031182004006390.
The developmental expression of the antigenic fucosylated glycan motifs Fucalpha1-3GalNAcbeta1-4GlcNAc (F-LDN), Fucalpha1-3GalNAcbeta1-4(Fucalpha1-3)GlcNAc (F-LDN-F), GalNAcbeta1-4(Fucalpha1-3)GlcNAc (LDN-F), Galbeta1-4(Fucalpha1-3)GlcNAc (Lewis X), and GalNAcbeta1-4(Fucalpha1-2Fucalpha1-3)GlcNAc (LDN-DF) in Schistosoma mansoni cercariae, adult worms and eggs, was surveyed using previously defined anti-carbohydrate monoclonal antibodies (mAbs). Lewis X was found both on glycolipids and glycoproteins, yet with completely different expression patterns during the life-cycle: on glycolipids, Lewis X was mainly found in the cercarial stage, while protein-conjugated Lewis X was mainly present in the egg stage. Also protein-conjugated LDN-F and LDN-DF were most highly expressed in the egg-stage. On glycolipids LDN-DF was found in all three examined stages, whereas LDN-F containing glycolipids were restricted to adult worms and eggs. The motifs F-LDN and F-LDN-F were found both on glycoproteins and glycolipids of the cercarial and egg stage, while in the adult stage, they appeared to occur predominantly on glycolipids. Immunofluorescence assays (IFA) showed that these F-LDN and F-LDN-F containing glycolipids were localized in a yet undefined duct or excretory system of adult worms. Murine infection serum showed major reactivity with this adult worm duct-system, which could be fully inhibited by pre-incubation with keyhole limpet haemocyanin (KLH). Clearly, the use of defined mAbs provides a quick and convenient way to map expression profiles of carbohydrate epitopes.
使用先前定义的抗碳水化合物单克隆抗体(mAb),对曼氏血吸虫尾蚴、成虫和虫卵中抗原性岩藻糖基化聚糖基序Fucα1-3GalNAcβ1-4GlcNAc(F-LDN)、Fucα1-3GalNAcβ1-4(Fucα1-3)GlcNAc(F-LDN-F)、GalNAcβ1-4(Fucα1-3)GlcNAc(LDN-F)、Galβ1-4(Fucα1-3)GlcNAc(Lewis X)和GalNAcβ1-4(Fucα1-2Fucα1-3)GlcNAc(LDN-DF)的发育表达进行了研究。发现Lewis X存在于糖脂和糖蛋白上,但在生命周期中具有完全不同的表达模式:在糖脂上,Lewis X主要存在于尾蚴阶段,而与蛋白质结合的Lewis X主要存在于虫卵阶段。同样,与蛋白质结合的LDN-F和LDN-DF在虫卵阶段表达最高。在糖脂上,LDN-DF在所有三个检测阶段均有发现,而含有LDN-F的糖脂仅限于成虫和虫卵。F-LDN和F-LDN-F基序在尾蚴和虫卵阶段的糖蛋白和糖脂上均有发现,而在成虫阶段,它们似乎主要存在于糖脂上。免疫荧光分析(IFA)表明,这些含有F-LDN和F-LDN-F的糖脂定位于成虫尚未明确的管道或排泄系统中。鼠感染血清与该成虫管道系统有主要反应性,预先与匙孔血蓝蛋白(KLH)孵育可完全抑制这种反应性。显然,使用定义明确的mAb为绘制碳水化合物表位的表达谱提供了一种快速便捷的方法。