Specific recombinant proteins of porcine epidemic diarrhea virus are immunogenic, revealing their potential use as diagnostic markers
Specific recombinant proteins of porcine epidemic diarrhea virus are immunogenic, revealing their potential use as diagnostic markers
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猪流行性腹泻病毒的特异性重组蛋白具有免疫原性，揭示了其作为诊断标记物的潜在用途
DOI：
10.1016/j.vetmic.2019.108387
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发表时间：
2019-09-01
影响因子：
3.3
通讯作者：
Huang, Yao-Wei
中科院分区：
文献类型：
--
作者：
Lei, Xi-Mei;Yang, Yong-Le;Huang, Yao-Wei
鉴于猪流行性腹泻（PED）具有高度传染性和急性发作的特点，尤其是对仔猪而言，迫切需要开发快速、灵敏的诊断检测方法。目前，猪流行性腹泻病毒（PEDV）特定辅助蛋白和非结构蛋白的诊断潜力（若存在的话）尚未得到研究。为了确定并比较哪些病毒蛋白可用作诊断抗原，我们通过酶联免疫吸附试验（ELISA）和/或蛋白质免疫印迹分析，检测了全病毒（WV）颗粒以及一组在细菌和/或哺乳动物细胞中单独表达的PEDV结构蛋白和非结构蛋白[刺突蛋白亚基1（S1）、开放阅读框3（ORF3）的C端部分（ORF3C）、包膜蛋白（E）、非结构蛋白1（Nsp1）、Nsp2、Nsp3的酸性结构域（Ac）以及Nsp3的ADP - 核糖 - 1 - 单磷酸酶结构域（ADRP）]与PEDV感染猪血清的反应性。蛋白质免疫印迹结果显示，血清抗体与S1蛋白的相互作用相较于ORF3C、E和Ac更为灵敏和特异。此外，我们通过ELISA分析了851份不同年龄腹泻猪的血清样本，大多数样本对WV、S1、ORF3C和E蛋白呈现免疫反应。最早在一周龄仔猪中观察到IgG抗体应答，S1、ORF3C、E和WV抗原的抗体发生过程和血清转化模式相似。另外，PEDV感染后，断奶仔猪的中和抗体模式与IgA更为相似。总体而言，这些数据为宿主对不同病毒蛋白的免疫反应提供了更可靠的信息，这将有助于开发新型血清学检测方法以及设计能更好激发保护性免疫的疫苗。
Given the highly contagious and acute nature of porcine epidemic diarrhea (PED), especially in piglets, there is an urgent need for the development of rapid and sensitive diagnostic assays. The diagnostic potentials of specific porcine epidemic diarrhea virus (PEDV) accessory and nonstructural proteins, if any, have not yet been investigated. In order to determine and compare which of the viral proteins may be useful as diagnostic antigens, whole virus (WV) particles and a panel of structural and nonstructural PEDV proteins [spike subunit 1 (S1), the C-terminal part of ORF3 (ORF3C), envelope (E), nonstructural protein 1 (Nsp1), Nsp2, Ac (acidic domain of Nsp3), and ADRP (ADP-ribose-1-monophosphatase domain of Nsp3), expressed individually in bacterial and/or mammalian cells] were tested for reactivity with sera from PEDV-infected pigs by ELISA and/or western blot analysis. According to western blots, serum antibody interactions with the S1 protein were relatively more sensitive and specific than ORF3C, E and Ac. Furthermore, a total of 851 serum samples from diarrheal pigs of different ages were analyzed by ELISA, with most showing immune-reactivity towards the WV, S1, ORF3C, and E proteins. The earliest IgG antibody response was observed in the one-week-old piglets, with similar antibody ontogeny and patterns of seroconversion for S1, ORF3C, E, and WV antigens. In addition, the pattern of neutralizing antibody was more similar to that of IgA in weaning piglets after PEDV infection. Collectively, these data provide more reliable information on the host immune response to different viral proteins, which will be useful for development of novel serological assays and for design of vaccines that better stimulate protective immunity.