Zhang Zhenchao, Wang Shuai, Li Charles, Liu Liheng
School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang, Henan, 453003, PR China.
Animal Biosciences and Biotechnology Laboratory, Agricultural Research Service, United States Department of Agriculture, Beltsville, MD 20705, USA.
Parasite. 2017;24:48. doi: 10.1051/parasite/2017047. Epub 2017 Dec 1.
The apicomplexan protozoans Eimeria spp. cause coccidioses, the most common intestinal diseases in chickens. Coccidiosis is associated with significant animal welfare issues and has a high economic impact on the poultry industry. Lack of a full understanding of immunogenic molecules and their precise functions involved in the Eimeria life cycles may limit development of effective vaccines and drug therapies. In this study, immunoproteomic approaches were used to define the antigenic protein repertoire from the total proteins of unsporulated Eimeria tenella oocysts. Approximately 101 protein spots were recognized in sera from chickens infected experimentally with E. tenella. Forty-six spots of unsporulated oocysts were excised from preparative gels and identified by matrix-assisted laser desorption ionization time-of-flight MS (MALDI-TOF-MS) and MALDI-TOF/TOF-MS. For unsporulated oocysts, 13 known proteins of E. tenella and 17 homologous proteins to other apicomplexan or protozoan parasites were identified using the 'Mascot' server. The remaining proteins were searched against the E. tenella protein sequence database using the 'Mascot in-house' search engine (version 2.1) in automated mode, and 12 unknown proteins were identified. The amino acid sequences of the unknown proteins were searched using BLAST against non-redundant sequence databases (NCBI), and 9 homologous proteins in unsporulated oocyst were found homologous to proteins of other apicomplexan parasites. These findings may provide useful evidence for understanding parasite biology, pathogenesis, immunogenicity and immune evasion mechanisms of E. tenella.
顶复门原生动物艾美耳球虫属会引发球虫病,这是鸡类中最常见的肠道疾病。球虫病与重大的动物福利问题相关,且对家禽业有着巨大的经济影响。对艾美耳球虫生命周期中免疫原性分子及其精确功能缺乏全面了解,可能会限制有效疫苗和药物疗法的研发。在本研究中,采用免疫蛋白质组学方法从未孢子化的柔嫩艾美耳球虫卵囊中定义抗原蛋白库。在用柔嫩艾美耳球虫进行实验性感染的鸡的血清中,大约识别出101个蛋白点。从未孢子化卵囊的制备凝胶中切下46个点,通过基质辅助激光解吸电离飞行时间质谱(MALDI - TOF - MS)和MALDI - TOF/TOF - MS进行鉴定。对于未孢子化卵囊,使用“Mascot”服务器鉴定出13种已知的柔嫩艾美耳球虫蛋白和17种与其他顶复门或原生动物寄生虫同源的蛋白。其余蛋白使用“Mascot内部”搜索引擎(版本2.1)在自动模式下针对柔嫩艾美耳球虫蛋白序列数据库进行搜索,鉴定出12种未知蛋白。使用BLAST针对非冗余序列数据库(NCBI)搜索未知蛋白的氨基酸序列,发现未孢子化卵囊中9种同源蛋白与其他顶复门寄生虫的蛋白同源。这些发现可能为理解柔嫩艾美耳球虫的寄生虫生物学特性、发病机制、免疫原性和免疫逃避机制提供有用证据。