Identification and characterization of cross-reactive antigens from Neospora caninum and Toxoplasma gondii
Identification and characterization of cross-reactive antigens from Neospora caninum and Toxoplasma gondii
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DOI：
10.1017/s0031182004006948
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发表时间：
2005-05-01
期刊：
影响因子：
2.4
通讯作者：
Fujisaki, K
中科院分区：
文献类型：
--
作者：
Liao, M;Xuan, X;Fujisaki, K
制备了针对犬新孢子虫速殖子的鼠单克隆抗体 (mAb)，以鉴定犬新孢子虫和弓形虫之间的交叉反应抗原。获得了识别两种寄生虫交叉反应抗原的 10 种单克隆抗体，并根据其在间接荧光抗体测试和蛋白质印迹分析中的反应模式初步分为 6 个不同的组。第一组中的三种单克隆抗体识别位于寄生虫表面的抗原，其分子量范围为 28 至 76 kDa；第 2 组中的一种 mAb 识别位于寄生虫内部细胞器上的抗原，分子量为 50 kDa；第 3 组中的一种 mAb 识别位于寄生虫内部细胞器上的抗原，分子量为 35 kDa 和 14 kDa；第 4 组中的三种 mAb 识别位于内部细胞器上、分子量为 64 kDa 的抗原；第 5 组中的一种 mAb 识别位于寄生虫表面的未知分子质量的抗原；第 6 组中的一种 mAb 识别位于寄生虫顶端的未知分子质量的抗原。第 1、2、3 和 5 组中的 mAb 在体外以剂量依赖性方式显示出对两种寄生虫生长的抑制作用。使用 mAb panel 对从犬新孢子虫速殖子 mRNA 制备的 cDNA 表达文库进行免疫筛选。蛋白质二硫键异构酶 (PDI)、热休克蛋白 70 (HSP70) 和核糖体蛋白 1 (RP1) 三种蛋白质分别被确定为第 2、3 和 4 组中 mAb 识别的交叉反应抗原。其中一些蛋白质可用于开发疫苗或药物来控制这两种寄生虫引起的疾病。
Murine monoclonal antibodies (mAbs) against Neospora caninum tachyzoites were produced to identify the cross-reactive antigens between N. caninum and Toxoplasma gondii. Ten mAbs recognizing cross-reactive antigens of both parasites were obtained and tentatively classified into 6 different groups based on their reactivity patterns in an indirect fluorescent antibody test and Western blot analysis. Three mAbs in group I recognized antigens located on the surface of parasites with molecular masses ranging from 28 to 76 kDa; one mAb in group 2 recognized antigens located on interior organelles of parasites with a molecular mass of 50 kDa; one mAb in group 3 recognized antigens located on interior organelles of parasites with molecular masses of 35 kDa and 14 kDa; three mAbs in group 4 recognized antigens located on interior organelles with a molecular mass of 64 kDa; one mAb in group 5 recognized antigens located on the surface of parasites with an unknown molecular mass; one mAb in group 6 recognized antigens located on the apical end of parasites with an unknown molecular mass. The mAbs in groups 1, 2, 3, and 5 showed inhibitory effects on the growth of the two parasites in vitro in a dose-dependent manner. A cDNA expression library prepared from N. caninum tachyzoite mRNA was immunoscreened with the mAb panel. Three kinds of proteins, protein disulfide isomerase (PDI), heat-shock protein 70 (HSP70), and ribosomal protein 1 (RP1), were identified as cross-reactive antigens recognized by mAbs in groups 2, 3, and 4, respectively. Some of the proteins could be useful in developing vaccines or drugs for controlling the diseases caused by the two parasites.