重组猪流行性腹泻病毒 Nsp9 的表达及其免疫原性。

Expression and immunogenicity of recombinant porcine epidemic diarrhea virus Nsp9.

机构信息

Institute of Pathogenic Microorganism and College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang, Jiangxi, China; Nanchang Key Laboratory of Animal Virus and Genetic Engineering, Jiangxi Agricultural University, Nanchang, 330045, China.

出版信息

Virology. 2023 Oct;587:109861. doi: 10.1016/j.virol.2023.109861. Epub 2023 Aug 3.

DOI:10.1016/j.virol.2023.109861

PMID:37572518

Abstract

Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea, vomiting, dehydration, and high mortality in newborn piglets, which leads to significant economic losses. Coronavirus nonstructural protein 9 (Nsp9) is an essential RNA binding protein for coronavirus replication, which renders it a promising candidate for developing antiviral drugs and diagnosis targeting PEDV. In this study, PEDV Nsp9 protein fused with MBP protein and His-tag were expressed and purified in Escherichia coli. Furthermore, immunization of MBP-Nsp9 enhances both humoral and cellular immunity responses as compared with that of His-Nsp9 protein. Finally, the swine immunization showed that Nsp9 protein could stimulate the swine immunity system to carry out humoral immunity, and the generated antibody could inhibit the proliferation of PEDV in Vero cells. Altogether, our data provide direct evidence for the immunogenicity of PEDV Nsp9, which sheds light on the future developments of anti-PEDV drugs and vaccines for PED prevention.

摘要

猪流行性腹泻病毒（PEDV）可引起新生仔猪急性腹泻、呕吐、脱水和高死亡率，给养殖业造成了巨大的经济损失。冠状病毒非结构蛋白 9（Nsp9）是冠状病毒复制所必需的 RNA 结合蛋白，使其成为开发抗 PEDV 药物和诊断方法的有前途的候选物。在本研究中，将 PEDV Nsp9 蛋白与 MBP 蛋白和 His 标签融合在大肠杆菌中表达和纯化。此外，与 His-Nsp9 蛋白相比，MBP-Nsp9 免疫可增强体液和细胞免疫反应。最后，猪免疫试验表明，Nsp9 蛋白可刺激猪免疫系统产生体液免疫，所产生的抗体可抑制 PEDV 在 Vero 细胞中的增殖。总的来说，我们的数据为 PEDV Nsp9 的免疫原性提供了直接证据，为开发抗 PEDV 药物和预防 PED 的疫苗提供了新的思路。