Dickerson H W, Clark T G, Findly R C
Department of Medical Microbiology, University of Georgia, Athens 30602.
J Protozool. 1989 Mar-Apr;36(2):159-64. doi: 10.1111/j.1550-7408.1989.tb01065.x.
Sera from fish that survive infections with the ciliated protozoon, Ichthyophthirius multifiliis, immobilize the parasite in vitro. In order to identify cell surface antigens involved in the immobilization response, integral membrane proteins were extracted from tomites with Triton X-114 and used to immunize rabbits. The rabbit antisera immobilized the parasite in vitro and antigens were localized to cell and ciliary plasma membranes by indirect immunofluorescent microscopy. The membrane protein fractions from both whole cells and tomite cilia were characterized by 1- and 2-dimensional SDS-PAGE. A 43,000-dalton (D) glycoprotein with an isoelectric point of 7.0 is the predominant protein in these fractions, comprising 12% and 60% of the total protein of whole cell and ciliary membranes, respectively. Western blot analysis of ciliary proteins with immune rabbit sera indicated that the 43,000-D glycoprotein is the principal antigen.
感染纤毛虫多子小瓜虫后存活下来的鱼类血清,能在体外使该寄生虫失去活动能力。为了鉴定参与这种固定反应的细胞表面抗原,用Triton X - 114从包囊幼虫中提取整合膜蛋白,并用于免疫兔子。兔抗血清在体外使寄生虫失去活动能力,通过间接免疫荧光显微镜观察,抗原定位于细胞和纤毛质膜上。全细胞和包囊幼虫纤毛的膜蛋白组分通过一维和二维SDS - PAGE进行表征。一种等电点为7.0、分子量为43,000道尔顿(D)的糖蛋白是这些组分中的主要蛋白质,分别占全细胞膜和纤毛膜总蛋白的12%和60%。用免疫兔血清对纤毛蛋白进行的蛋白质印迹分析表明,43,000 - D糖蛋白是主要抗原。