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携带类NADC30猪繁殖与呼吸综合征病毒样颗粒共表达的重组伪狂犬病病毒的构建及免疫原性分析

The Construction and Immunogenicity Analyses of Recombinant Pseudorabies Virus With NADC30-Like Porcine Reproductive and Respiratory Syndrome Virus-Like Particles Co-expression.

作者信息

Zhao Jun, Zhu Ling, Xu Lei, Li Fengqing, Deng Huidan, Huang Yao, Gu Sirui, Sun Xianggang, Zhou Yuancheng, Xu Zhiwen

机构信息

College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China.

Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China.

出版信息

Front Microbiol. 2022 Mar 2;13:846079. doi: 10.3389/fmicb.2022.846079. eCollection 2022.

DOI:10.3389/fmicb.2022.846079
PMID:35308386
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8924499/
Abstract

Porcine reproductive and respiratory syndrome (PRRS) and pseudorabies (PR) are highly infectious swine diseases and cause significant financial loss in China. The respiratory system and reproductive system are the main target systems. Previous studies showed that the existing PR virus (PRV) and PRRS virus (PRRSV) commercial vaccines could not provide complete protection against PRV variant strains and NADC30-like PRRSV strains in China. In this study, the PRV variant strain XJ and NADC30-like PRRSV strain CHSCDJY-2019 are used as the parent for constructing a recombinant pseudorabies virus (rPRV)-NC56 with gE/gI/TK gene deletion and co-expressing NADC30-like PRRSV GP5 and M protein. The rPRV-NC56 proliferated stably in BHK-21 cells, and it could stably express GP5 and M protein. Due to the introduction of the self-cleaving 2A peptide, GP5 and M protein were able to express independently and form virus-like particles (VLPs) of PRRSV in rPRV-NC56-infected BHK-21 cells. The rPRV-NC56 is safe for use in mice; it can colonize and express the target protein in mouse lungs for a long time. Vaccination with rPRV-NC56 induces PRV and NADC30-like PRRSV specific humoral and cellular immune responses in mice, and protects 100% of mice from virulent PRV XJ strain. Furthermore, the virus-neutralizing antibody (VNA) elicited by rPRV-NC56 showed significantly lower titer against SCNJ-2016 (HP-PRRSV) than that against CHSCDJY-2019 (NADC30-like PRRSV). Thus, rPRV-NC56 appears to be a promising candidate vaccine against NADC30-like PRRSV and PRV for the control and eradication of the variant PRV and NADC30-like PRRSV.

摘要

猪繁殖与呼吸综合征（PRRS）和伪狂犬病（PR）是高度传染性的猪病，在中国造成了巨大的经济损失。呼吸系统和生殖系统是主要的靶系统。先前的研究表明，现有的伪狂犬病病毒（PRV）和猪繁殖与呼吸综合征病毒（PRRSV）商业疫苗无法对中国的PRV变异株和NADC30样PRRSV株提供完全保护。在本研究中，以PRV变异株XJ和NADC30样PRRSV株CHSCDJY - 2019为亲本，构建了缺失gE/gI/TK基因并共表达NADC30样PRRSV GP5和M蛋白的重组伪狂犬病病毒（rPRV）- NC56。rPRV - NC56在BHK - 21细胞中稳定增殖，且能稳定表达GP5和M蛋白。由于引入了自我切割的2A肽，GP5和M蛋白能够独立表达，并在rPRV - NC56感染的BHK - 21细胞中形成PRRSV病毒样颗粒（VLP）。rPRV - NC56对小鼠使用安全；它可以在小鼠肺中定殖并长时间表达靶蛋白。用rPRV - NC56免疫可诱导小鼠产生PRV和NADC30样PRRSV特异性体液免疫和细胞免疫应答，并能100%保护小鼠免受强毒PRV XJ株的攻击。此外，rPRV - NC56诱导产生的病毒中和抗体（VNA）针对SCNJ - 2016（HP - PRRSV）的效价比针对CHSCDJY - 2019（NADC30样PRRSV）的效价显著降低。因此，rPRV - NC56似乎是一种有前景的候选疫苗，可用于防控和根除变异PRV和NADC30样PRRSV。

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