National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-Cho, Obihiro, Hokkaido 080-8555, Japan; Department of Animal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, University of Sadat City, Minoufiya 32897, Egypt.
Exp Parasitol. 2013 Nov;135(3):623-8. doi: 10.1016/j.exppara.2013.09.017. Epub 2013 Sep 30.
Apical membrane antigen-1 (AMA-1) is a microneme protein that exists in all apicomplexan parasites and plays an indispensable role in the invasion into host cell. Central region of ectodomains I and II of Babesia bovis apical membrane antigen-1 (BbAMA-1P) is highly conserved with these of Babesia species and may be beneficial for vaccine development against babesiosis. In the present study, recombinant protein encoding the central region of B. bovis AMA-1 (rBbAMA-1P) was produced in Escherichia coli and its antiserum was prepared in mice for further molecular characterization. Anti-rBbAMA-1P serum specifically reacted with corresponding authentic protein of B. bovis as determined by Western blotting and IFAT. Cultured B. bovis treated with anti-rBbAMA-1P serum showed significant reduction in the in vitro growth of the parasites. Moreover, preincubated free merozoites with 1mg/ml anti-rBbAMA-1P serum inhibited their efficiency in the invasion into erythrocytes (RBCs) by 61% and 70% at 3h and 6h, respectively. Our data suggest that the central region of domains I and II of BbAMA-1 may serve as a vaccine candidate against babesiosis.
顶膜蛋白-1（AMA-1）是一种存在于所有顶复体寄生虫中的微线蛋白，在入侵宿主细胞中起着不可或缺的作用。牛巴贝斯虫顶膜抗原-1（BbAMA-1P）的结构域 I 和 II 的外区中心区域与这些巴贝斯虫物种高度保守，可能有利于开发抗巴贝斯虫病疫苗。在本研究中，在大肠杆菌中产生了编码牛顶膜 AMA-1 的中心区域的重组蛋白，并在小鼠中制备了针对该蛋白的抗血清，用于进一步的分子特征分析。Western blot 和 IFAT 分析表明，抗 rBbAMA-1P 血清特异性地与牛巴贝斯虫的相应真实蛋白反应。用抗 rBbAMA-1P 血清处理培养的牛巴贝斯虫后，寄生虫在体外的生长明显减少。此外，用 1mg/ml 的抗 rBbAMA-1P 血清预先孵育游离的裂殖子，分别在 3h 和 6h 时将其入侵红细胞（RBC）的效率降低了 61%和 70%。我们的数据表明，BbAMA-1 的结构域 I 和 II 的中心区域可能是抗巴贝斯虫病疫苗的候选物。
