Messenger RNA-based vaccines represent new tools with prophylactic and therapeutic potential characterized by high flexibility of application for infectious diseases. Pseudorabies virus (PRV) is one of the major viruses affecting the pig industry. PRV has serious effects in piglets, sows, and growing-fattening pigs and can lead to huge economic losses. In this study, an envelope glycoprotein D (gD) gene-based specific mRNA vaccine was generated, and a mouse model was used to investigate the protective efficacy of the vaccine. The gD mRNA vaccine and the recombinant plasmid pVAX-gD were transfected into BHK21 cells, and the antigenicity of the expressed proteins was detected by Western blot analysis. Groups of mice were vaccinated with the gD mRNA vaccine, pVAX-gD, and PBS. T cell immune responses were measured by flow cytometry or ELISA and serum neutralization tests every two weeks. The challenge with the PRV-XJ strain was performed eight weeks after the primary immunization, and the response was monitored for 15 days. The levels of specific and neutralizing antibodies in the gD mRNA vaccine group were significantly increased in 8 weeks compared to those in the control group, and cytokine levels, including that of IFN-γ/IL-2, were considerably higher than those in the control animal. Additionally, the proportion of CD4⁺/CD8⁺ cells in peripheral lymphocytes was remarkably increased. Our data demonstrate that mRNA is a promising and effective tool for the development of vaccines. The PRV-gD-based mRNA vaccine can elicit an efficient neutralizing antibody response and induce effective protection in mice in defense against PRV infection.

基于信使RNA的疫苗代表了具有预防和治疗潜力的新工具，其特点是对传染病的应用具有高度灵活性。伪狂犬病病毒（PRV）是影响养猪业的主要病毒之一。 PRV 对仔猪、母猪和生长育肥猪有严重影响，可导致巨大的经济损失。在这项研究中，产生了一种基于包膜糖蛋白D（gD）基因的特异性mRNA疫苗，并利用小鼠模型研究了该疫苗的保护功效。将gD mRNA疫苗和重组质粒pVAX-gD转染BHK21细胞，Western blot分析检测表达蛋白的抗原性。各组小鼠分别接种 gD mRNA 疫苗、pVAX-gD 和 PBS。每两周通过流式细胞术或 ELISA 以及血清中和试验测量 T 细胞免疫反应。初次免疫后八周进行 PRV-XJ 毒株的攻击，并监测反应 15 天。与对照组相比，gD mRNA疫苗组的特异性抗体和中和抗体水平在8周内显着升高，细胞因子水平，包括IFN-γ/IL-2，显着高于对照组动物。另外，外周淋巴细胞中CD4 ⁺ /CD8 ⁺细胞的比例显着增加。我们的数据表明 mRNA 是疫苗开发的一种有前途且有效的工具。基于 PRV-gD 的 mRNA 疫苗可以引发有效的中和抗体反应，并诱导小鼠有效防御 PRV 感染。