Sun Leqiang, Tang Yajie, Yan Keji, Chen Huanchun, Zhang Huawei
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070 Hubei China.
Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070 Hubei China.
Anim Dis. 2021;1(1):22. doi: 10.1186/s44149-021-00021-9. Epub 2021 Oct 8.
Porcine epidemic diarrhea (PED) caused by the porcine epidemic diarrhea virus (PEDV), is a severe infectious and devastating swine disease that leads to serious economic losses in the swine industry worldwide. An increased number of PED cases caused by variant PEDV have been reported in many countries since 2010. S protein is the main immunogenic protein containing some B-cell epitopes that can induce neutralizing antibodies of PEDV. In this study, the construction, expression and purification of Pseudomonas aeruginosa exotoxin A (PE) without domain III (PEΔIII) as a vector was performed for the delivery of PEDV S-A or S-B. PE(ΔIII) PEDV S-A and PE(ΔIII) PEDV S-B recombinant proteins were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis. The immunogenicity of PEDV S-A and PEDV S-B subunit vaccines were evaluated in mice. The results showed that PEDV-S-B vaccine could not only induce specific humoral and Th1 type-dominant cellular immune responses, but also stimulate PEDV-specific mucosal immune responses in mice. PEDV-S-B subunit vaccine is a novel candidate mucosal vaccine against PEDV infection.
由猪流行性腹泻病毒(PEDV)引起的猪流行性腹泻(PED)是一种严重的传染性和毁灭性猪病,在全球养猪业中导致严重的经济损失。自2010年以来,许多国家报告了由变异PEDV引起的PED病例数量增加。S蛋白是主要的免疫原性蛋白,含有一些可诱导PEDV中和抗体的B细胞表位。在本研究中,构建、表达并纯化了无结构域III的铜绿假单胞菌外毒素A(PE)作为载体,用于递送PEDV S-A或S-B。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白质免疫印迹分析证实了PE(ΔIII) PEDV S-A和PE(ΔIII) PEDV S-B重组蛋白。在小鼠中评估了PEDV S-A和PEDV S-B亚单位疫苗的免疫原性。结果表明,PEDV-S-B疫苗不仅能诱导特异性体液免疫和以Th1型为主的细胞免疫反应,还能刺激小鼠产生PEDV特异性黏膜免疫反应。PEDV-S-B亚单位疫苗是一种新型的抗PEDV感染黏膜疫苗候选物。