Shin H J, Kim C H, Im K I
Department of Biology, College of Natural Science, Chungnam National University, Taejon, Korea.
Kisaengchunghak Chapchi. 1992 Dec;30(4):289-98. doi: 10.3347/kjp.1992.30.4.289.
Acanthamoeba spp., free-living amoebae inhabited in moist soil, pond, freshwater, sewage, atmosphere and swimming pool, may be causative protozoa of the fatal primary amoebic meningoencephalitis in experimental animals and humans. In this study, Acanthamoeba spp., including Acanthamoeba sp. YM-4 (isolated strain from Korea) had been compared by the two-dimensional electrophoresis and hybridoma technique as well as the difference of morphological characteristics. Trophozoite of Acanthamoeba sp. YM-4 is usually uninucleate and show the hyaline filamentous projections (acanthopoda). No flagellate stage observed. Cysts have two walls, the outer wall is nearly circular, but inner wall is oval or some irregular. As results of SDS-PAGE for lysate of Acanthamoeba sp. YM-4, 16 major protein fractions are similar to those of A. culbertsoni, but different to A. royreba and A. polyphaga. Findings of two-dimensional electrophoretic patterns of Acanthamoeba sp. YM-4 are almost same to those of A. culbertsoni, The isotype of monoclonal antibodies produced from McAY 6, McAY 7, McAY 8, McAY 13 and McAY 16 clones were IgG1, and McAY 10 and McAY 11 clones were IgM. As results of the cross-reactivity among various amoebae using ELISA with monoclonal antibodies, McAY 7 monoclonal antibody (molecular weight 43 kDa by EITB) was only reacted with Acanthamoeba sp. YM-4, but McAY 6 and McAY 10 monoclonal antibodies were reacted to A. culbertsoni as well as Acanthamoeba sp. YM-4.
棘阿米巴属是一种自由生活的变形虫,栖息于潮湿土壤、池塘、淡水、污水、空气和游泳池中,可能是实验动物和人类致命性原发性阿米巴脑膜脑炎的致病原虫。在本研究中,通过二维电泳、杂交瘤技术以及形态学特征差异,对包括棘阿米巴属YM-4菌株(从韩国分离得到)在内的棘阿米巴属进行了比较。棘阿米巴属YM-4菌株的滋养体通常为单核,具有透明的丝状伪足(棘状伪足)。未观察到鞭毛阶段。包囊有两层壁,外壁近圆形,但内壁为椭圆形或有些不规则。对棘阿米巴属YM-4菌株裂解物进行SDS-PAGE的结果显示,16个主要蛋白组分与库氏棘阿米巴的相似,但与罗氏棘阿米巴和多食棘阿米巴不同。棘阿米巴属YM-4菌株的二维电泳图谱结果与库氏棘阿米巴几乎相同,从McAY 6、McAY 7、McAY 8、McAY 13和McAY 16克隆产生的单克隆抗体的同种型为IgG1,而McAY 10和McAY 11克隆为IgM。使用单克隆抗体通过ELISA对各种阿米巴之间的交叉反应性进行检测的结果显示,McAY 7单克隆抗体(EITB检测分子量为43 kDa)仅与棘阿米巴属YM-4菌株反应,但McAY 6和McAY 10单克隆抗体与库氏棘阿米巴以及棘阿米巴属YM-4菌株均有反应。