Key Laboratory of Bio-Resources and Eco-Environment, Ministry of Education, College of Life Science. Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, Chengdu 610064, China.
Key Laboratory of Bio-Resources and Eco-Environment, Ministry of Education, College of Life Science. Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, Chengdu 610064, China; Sichuan Animal Science Academy (SASA), Chengdu 610066, China.
Vet Microbiol. 2024 Nov;298:110241. doi: 10.1016/j.vetmic.2024.110241. Epub 2024 Aug 30.
Porcine rotavirus (PoRV), a member of the Reoviridae family, constitutes a principal etiological agent of acute diarrhea in piglets younger than eight weeks of age, and it is associated with considerable morbidity and mortality within the swine industry. The G5 genotype rotavirus strain currently predominates in circulation. To develop a safe and effective porcine rotavirus vaccine, we generated an insect cell-baculovirus expression system, and successfully expressed these three viral proteins and assembled them into virus-like particles (VLPs) co-displaying VP2, VP6, and VP7. Transmission electron microscopy (TEM) analysis revealed that the VP2-VP6-VP7 VLPs exhibited a "wheeled" morphology resembling that of native rotavirus particles, with an estimated diameter of approximately 65 nm. To evaluate the immunogenicity and protective efficacy of these VP2-VP6-VP7 VLPs, we immunized BALB/C mice with four escalating doses of the VLPs, ranging from 5 to 40 μg of VLP protein per dose. ELISA-based assessments of PoRV-specific antibodies and T cell cytokines, including IL-4, IL-2, and IFN-γ, demonstrate that immunization with VP2-VP6-VP7 VLPs can effectively elicit both humoral and cellular immune responses in mice, resulting in a notable induction of neutralizing antibodies. On days 4, 6, 8, and 10 post-infection (dpi), the VLP-vaccinated group exhibited significantly reduced levels of PoRV RNA copy numbers when compared to the PBS controls. Histological examination of the duodenum, ileum, and kidneys revealed that VP2-VP6-VP7 VLPs provided effective protection against PoRV induced intestinal injury. Collectively, these findings indicate that the VLPs generated in this study possess strong immunogenicity and suggest the considerable promise of the VLP-based vaccine candidate in the prevention and containment of Porcine Rotavirus infections.
猪轮状病毒(PoRV)是呼肠孤病毒科的一个成员,是 8 周龄以下仔猪急性腹泻的主要病因之一,它与养猪业的高发病率和死亡率密切相关。目前,G5 基因型轮状病毒株在循环中占主导地位。为了开发安全有效的猪轮状病毒疫苗,我们构建了昆虫细胞-杆状病毒表达系统,成功表达了这三种病毒蛋白,并将它们组装成共同展示 VP2、VP6 和 VP7 的病毒样颗粒(VLPs)。透射电子显微镜(TEM)分析显示,VP2-VP6-VP7 VLPs 呈现出类似于天然轮状病毒颗粒的“轮状”形态,直径约为 65nm。为了评估这些 VP2-VP6-VP7 VLPs 的免疫原性和保护效力,我们用 4 个递增剂量的 VLPs(每个剂量 5 到 40μg 的 VLP 蛋白)对 BALB/C 小鼠进行免疫。ELISA 法检测 PoRV 特异性抗体和 T 细胞细胞因子(包括 IL-4、IL-2 和 IFN-γ)表明,VP2-VP6-VP7 VLPs 免疫可有效诱导小鼠产生体液和细胞免疫应答,显著诱导中和抗体。在感染后第 4、6、8 和 10 天(dpi),与 PBS 对照组相比,VP2-VP6-VP7 VLPs 免疫组的 PoRV RNA 拷贝数显著降低。对十二指肠、回肠和肾脏的组织学检查表明,VP2-VP6-VP7 VLPs 可有效防止 PoRV 引起的肠道损伤。综上所述,这些结果表明,本研究中产生的 VLPs 具有很强的免疫原性,基于 VLP 的候选疫苗在预防和控制猪轮状病毒感染方面具有很大的应用前景。