Ni Junli, Chen Xiangjie, Sun Yunyun, Zhu Yibin, Cai Haiming, Song Yongle, Lv Minna, Li Juan, Liao Shenquan, Qi Nanshan, Sun Mingfei, Zhang Jianfei, Gu Youfang, Liu Xinchao, Lin Xuhui
Guangdong Province Key Laboratory of Livestock Disease Prevention, Key Laboratory of Avian Infuenza and Other Major Poultry Diseases Prevention and Control, Ministry of Agriculture and Rural Afairs, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China; College of Animal Science and Technology, Anhui Science and Technology University, Fengyang 233100, China.
Guangdong Province Key Laboratory of Livestock Disease Prevention, Key Laboratory of Avian Infuenza and Other Major Poultry Diseases Prevention and Control, Ministry of Agriculture and Rural Afairs, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China.
Poult Sci. 2025 Feb;104(2):104721. doi: 10.1016/j.psj.2024.104721. Epub 2024 Dec 31.
Avian coccidiosis, caused by protozoan parasites of the genus Eimeria, is a globally prevalent and highly pathogenic disease that poses a serious threat to the poultry industry, resulting in significant economic losses. However, the mechanism by which Eimeria species invade host cells remains unclear. Previous studies have identified rhoptry neck protein 2 (RON2) from Eimeria tenella as a critical factor in host cell invasion, but a comprehensive understanding of the role of EtRON2 in host cell invasion and its relationship with E. tenella invasion is lacking. To address this gap, this study focused on the secreted protein EtRON2 from E. tenella and its interaction with host cell receptors. The receptor interacting with the EtRON2 protein was identified through a GST pull-down assay, followed by mass spectrometry, and the interaction was further validated through subcellular co-localization analysis. Furthermore, sporozoites and host cells were treated with a specific antibody targeting the EtRON2/annexin A2 interaction, and the invasion rate of E. tenella was assessed using RT-qPCR to analyze the effect of inhibiting this interaction on host cell invasion by E. tenella. Annexin A2 protein, located on the surface of the host cell membrane, was screened, and the results of the sporozoite invasion assay revealed that inhibiting the interaction between these two proteins reduced F-actin aggregation. Understanding the interaction between the EtRON2 protein and its receptor during parasite invasion could help elucidate the function of the EtRON2 receptor and provide a theoretical foundation for further studies on the invasion mechanisms of E. tenella and the prevention and control of avian coccidiosis.
禽球虫病由艾美耳属原生动物寄生虫引起，是一种全球流行且致病性很强的疾病，对家禽业构成严重威胁，导致重大经济损失。然而，艾美耳属物种侵入宿主细胞的机制仍不清楚。先前的研究已确定来自柔嫩艾美耳球虫的棒状体颈部蛋白2（RON2）是宿主细胞入侵的关键因素，但对柔嫩艾美耳球虫RON2（EtRON2）在宿主细胞入侵中的作用及其与柔嫩艾美耳球虫入侵的关系仍缺乏全面了解。为填补这一空白，本研究聚焦于柔嫩艾美耳球虫分泌蛋白EtRON2及其与宿主细胞受体的相互作用。通过谷胱甘肽S-转移酶（GST）下拉试验鉴定与EtRON2蛋白相互作用的受体，随后进行质谱分析，并通过亚细胞共定位分析进一步验证这种相互作用。此外，用靶向EtRON2/膜联蛋白A2相互作用的特异性抗体处理子孢子和宿主细胞，使用逆转录定量聚合酶链反应（RT-qPCR）评估柔嫩艾美耳球虫的入侵率，以分析抑制这种相互作用对柔嫩艾美耳球虫宿主细胞入侵的影响。筛选位于宿主细胞膜表面的膜联蛋白A2蛋白，子孢子入侵试验结果表明，抑制这两种蛋白之间的相互作用可减少F-肌动蛋白聚集。了解寄生虫入侵过程中EtRON2蛋白与其受体之间的相互作用，有助于阐明EtRON2受体的功能，并为进一步研究柔嫩艾美耳球虫的入侵机制及禽球虫病的防控提供理论基础。
