Harth G, Mills A A, Souto-Padrón T, de Souza W
Department of Immunology and Infectious Diseases, Palo Alto Medical Foundation, California 94301.
Mol Cell Biochem. 1992 Jan 15;109(1):25-36. doi: 10.1007/BF00230870.
Two monoclonal antibodies were used to biochemically characterize glycoprotein 72 (GP72) from Trypanosoma cruzi and to localize the protein in live and fixed parasites by indirect immunofluorescence and in thin section of parasites by immunogold electron microscopy. GP72 was shown in immunoblots to be specific for the epimastigote stage; the protein could not be detected in trypomastigotes. Each antibody reacted with a different epitope on the glycoprotein and deglycosylation of GP72 ablated reactivity with one of the antibodies. Indirect immunofluorescence and electron microscopic evaluation of parasite associated gold particles showed the presence of GP72 in the cell surface membrane including the flagellar pocket and the cytostome. In addition, cytoplasmic membrane vesicles of the endosomal-lysosomal system stained intensely.
使用两种单克隆抗体对克氏锥虫的糖蛋白72（GP72）进行生化特性分析，并通过间接免疫荧光法在活的和固定的寄生虫中定位该蛋白，通过免疫金电子显微镜在寄生虫的薄切片中定位该蛋白。免疫印迹显示GP72对前鞭毛体阶段具有特异性；在锥鞭毛体中未检测到该蛋白。每种抗体与糖蛋白上的不同表位发生反应，GP72的去糖基化消除了与其中一种抗体的反应性。对寄生虫相关金颗粒的间接免疫荧光和电子显微镜评估显示，GP72存在于细胞表面膜中，包括鞭毛袋和胞口。此外，内体-溶酶体系统的细胞质膜囊泡染色强烈。
