College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin 150030, China.
Virus Res. 2012 Aug;167(2):259-66. doi: 10.1016/j.virusres.2012.05.007. Epub 2012 May 19.
Porcine epidemic diarrhea virus (PEDV) is the causative agent of porcine epidemic diarrhea, a highly contagious enteric disease of swine. The Spike (S) protein is one of the main structural proteins of PEDV capable of inducing neutralizing antibodies in vivo. Herein, we generated three distinct DNA constructs in the eukaryotic expression plasmid pVAX1; one encoding the S protein [pVAX1-(PEDV-S)], the second encoding the N-terminal fragment (S1) [pVAX1-(PEDV-S1)] containing potent antigenic sites, and the third expressing the porcine interleukin-18 (pIL-18) [pVAX1-(IL-18)]. Immunofluorescence assays in BHK-21 cells demonstrated successful protein expression from all 3 constructs. Kunming mice were injected separately with each of these constructs or with a pVAX1-(PEDV-S1)/pVAX1-(IL-18) combination, an attenuated PEDV vaccine, or vector only control. Animals were examined for T lymphocyte proliferation, anti-PEDV antibodies, IFN-γ and IL-4 protein levels, and cytotoxic T cell function in mouse peripheral blood and spleen. In all cases, results showed that pVAX1-(PEDV-S) and the combination of pVAX1-(PEDV-S1) with pVAX1-(IL-18) induced the strongest responses; however, pIL-18 had no adjuvant effects when given in combination with pVAX1-(PEDV-S1).
猪流行性腹泻病毒(PEDV)是猪流行性腹泻的病原体,是一种高度传染性的猪肠道疾病。刺突(S)蛋白是 PEDV 的主要结构蛋白之一,能够在体内诱导中和抗体。在此,我们在真核表达质粒 pVAX1 中生成了三个不同的 DNA 构建体;一个编码 S 蛋白[pVAX1-(PEDV-S)],第二个编码包含有效抗原位点的 N 端片段(S1)[pVAX1-(PEDV-S1)],第三个表达猪白细胞介素-18(pIL-18)[pVAX1-(IL-18)]。BHK-21 细胞中的免疫荧光分析表明,所有 3 个构建体均成功表达了蛋白。昆明小鼠分别用这些构建体中的每一个或用减毒 PEDV 疫苗 pVAX1-(PEDV-S1)/pVAX1-(IL-18)组合、载体对照进行注射。检查动物的 T 淋巴细胞增殖、抗 PEDV 抗体、IFN-γ和 IL-4 蛋白水平以及小鼠外周血和脾脏中的细胞毒性 T 细胞功能。在所有情况下,结果表明 pVAX1-(PEDV-S)和 pVAX1-(PEDV-S1)与 pVAX1-(IL-18)的组合诱导了最强的反应;然而,当与 pVAX1-(PEDV-S1)联合使用时,pIL-18 没有佐剂作用。