Eimeria tenella Eimeria-specific protein that interacts with apical membrane antigen 1 (EtAMA1) is involved in host cell invasion

文献类型： 外文期刊

第一作者： Li, Cong

作者： Li, Cong;Zhao, Qiping;Zhu, Shunhai;Wang, Qingjie;Wang, Haixia;Yu, Shuilan;Yu, Yu;Liang, Shashan;Zhao, Huanzhi;Huang, Bing;Dong, Hui;Han, Hongyu;Yu, Yu;Liang, Shashan

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关键词： Eimeria tenella; Apical membrane antigen 1; Eimeria-specific protein

期刊名称：PARASITES & VECTORS （ 影响因子：3.876； 五年影响因子：3.959 ）

ISSN： 1756-3305

年卷期： 2020 年 13 卷 1 期

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收录情况： SCI

摘要： Background Avian coccidiosis is a widespread, economically significant disease of poultry, caused by severalEimeriaspecies. These parasites have complex and diverse life-cycles that require invasion of their host cells. This is mediated by various proteins secreted from apical secretory organelles. Apical membrane antigen 1 (AMA1), which is released from micronemes and is conserved across all apicomplexans, plays a central role in the host cell invasion. In a previous study, some putativeEtAMA1-interacting proteins ofE. tenellawere screened. In this study, we characterized one putativeEtAMA1-interacting protein,E. tenella Eimeria-specific protein (EtEsp). Methods Bimolecular fluorescence complementation (BiFC) and glutathione S-transferase (GST) fusion protein pull-down (GST pull-down) were used to confirm the interaction betweenEtAMA1 andEtEspin vivoandin vitro.The expression ofEtEsp was analyzed in different developmental stages ofE. tenellawith quantitative PCR and western blotting. The secretion ofEtEsp protein was tested with staurosporine when sporozoites were incubated in complete medium at 41 degrees C. The localization ofEtEsp was analyzed with an immunofluorescence assay (IFA). Anin vitroinvasion inhibition assay was conducted to assess the ability of antibodies againstEtEsp to inhibit cell invasion byE. tenellasporozoites. Results The interaction betweenEtAMA1 andEtEsp was confirmed with BiFC and by GST pull-down. Our results show thatEtEsp is differentially expressed during distinct phases of the parasite life-cycle. IFA showed that theEtEsp protein is mainly distributed on the parasite surface, and that the expression of this protein increases during the development of the parasite in the host cells. Using staurosporine, we showed thatEtEsp is a secreted protein, but not from micronemes. In inhibition tests, a polyclonal anti-rEtEsp antibody attenuated the capacity ofE. tenellato invade host cells. Conclusion In this study, we show thatEtEsp interacts withEtAMA1 and that the protein is secreted protein, but not from micronemes. The protein participates in sporozoite invasion of host cells and is maybe involved in the growth of the parasite. These data have implications for the use ofEtAMA1 orEtAMA1-interacting proteins as targets in intervention strategies against avian coccidiosis.

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