Deelder A M, van Dam G J, Kornelis D, Fillié Y E, van Zeyl R J
University of Leiden, Department of Parasitology, The Netherlands.
Parasitology. 1996 Jan;112 ( Pt 1):21-35. doi: 10.1017/s0031182000065045.
Using spleen cells of mice infected or immunized respectively with cercariae or antigen preparations of Schistosoma mansoni, S. haematobium or S. japonicum monoclonal antibodies (mAbs) were produced against the schistosome gut-associated antigens CAA (circulating anodic antigen) and CCA (circulating cathodic antigen). Fusions nearly exclusively produced either anti-CAA (n = 25) or anti-CCA mAbs (n = 55) with a strong isotype restriction (IgM, IgG1 and IgG3) against both antigens, the majority of anti-CAA mAbs being IgG1 and the majority of anti-CCA mAbs being IgM. The mAbs, which on the basis of their selection were reactive with multiple carbohydrate epitopes of CAA or CCA, were applied in different immunological techniques including immunofluorescence, a dot immunobinding assay and immunoelectrophoresis to study the epitope repertoire. Anti-CAA mAbs were found to be reactive with 5 different epitopes, none of which occurred as multiple epitopes on eggs. Anti-CCA mAbs, on the other hand, recognized at least 10 different epitopes, while 44% of anti-CCA mAbs recognized epitopes common to the adult worm and the egg. Both CAA- and CCA-epitopes were found to be developmentally expressed at the level of the tegument in cercariae, schistosomula and 5-day-old lung worms, but in the adult worm were primarily found in the gut. Thus, the production of panels of mAbs has not only resulted in the selection of reagents optimally performing in diagnostic immunoassays, but also allowed a more detailed study of the epitope repertoire of these important schistosome antigens.
分别用曼氏血吸虫、埃及血吸虫或日本血吸虫的尾蚴或抗原制剂感染或免疫小鼠,取其脾细胞制备针对血吸虫肠道相关抗原循环阳极抗原(CAA)和循环阴极抗原(CCA)的单克隆抗体(mAb)。融合几乎只产生抗CAA(n = 25)或抗CCA单克隆抗体(n = 55),对这两种抗原均有强烈的同种型限制(IgM、IgG1和IgG3),大多数抗CAA单克隆抗体为IgG1,大多数抗CCA单克隆抗体为IgM。基于其选择与CAA或CCA的多个碳水化合物表位反应的单克隆抗体,被应用于包括免疫荧光、斑点免疫结合试验和免疫电泳在内的不同免疫技术中,以研究表位库。发现抗CAA单克隆抗体与5种不同表位反应,其中没有一种在虫卵上以多个表位形式出现。另一方面,抗CCA单克隆抗体识别至少10种不同表位,而44%的抗CCA单克隆抗体识别成虫和虫卵共有的表位。发现CAA和CCA表位在尾蚴、童虫和5日龄肺期虫体的体表水平上呈发育性表达,但在成虫中主要存在于肠道。因此,单克隆抗体组的产生不仅导致了在诊断免疫测定中性能最佳的试剂的选择,而且还允许对这些重要血吸虫抗原的表位库进行更详细的研究。