Li Xunliang, Li Pengchong, Cao Liyan, Bai Yunyun, Chen Huijie, Liu He, Ren Xiaofeng, Li Guangxing
College of Veterinary Medicine, Key Laboratory for Laboratory Animals and Comparative Medicine of Heilongjiang Province, Northeast Agricultural University, 600 Changjiang Road, Xiangfang District, Harbin 150030, China.
Fushun Committee of Agriculture, East of Linjiang Street, Shuncheng District, Fushun 113006, China.
J Vet Med Sci. 2019 Oct 18;81(10):1438-1444. doi: 10.1292/jvms.18-0682. Epub 2019 Sep 2.
Transmissible gastroenteritis (TGE), caused by transmissible gastroenteritis virus (TGEV), is a highly infectious disease in pigs. Vaccination is an effective approach to prevent TGEV infection. Here, we evaluated the potential of TGEV S1 as a DNA vaccine and porcine interleukin (pIL)-12 as an adjuvant in a mouse model. A DNA vaccine was constructed with the TGEV S1 gene to induce immune response in an experimental mouse model; pIL-12 was chosen as the immunological adjuvant within this DNA vaccine. The pVAX1-(TGEV-S1) and pVAX1-(pIL-12) vectors were transfected into BHK-21 cells and expressed in vitro. Experimental mice were separately immunized with each of the recombinant plasmids and controls through the intramuscular route. The lymphocytes isolated from the blood and spleen were analyzed for proliferation, cytotoxic activities, and populations of CD4 and CD8 cells. The titers of TGEV S1 in an enzyme-linked immunosorbent assay (ELISA) and TGEV neutralizing antibodies and the concentrations of interferon (IFN)-γ and IL-4 were also analyzed in the serum. The plasmids pVAX1-(TGEV-S1) and pVAX1-(pIL-12) could be expressed in BHK-21 cells, and the combination of pVAX1-(TGEV-S1) and pVAX1-(pIL-12) could induce a significant increase in all markers. pIL-12 could act as an immunological adjuvant in the DNA vaccine for TGEV-S1. Furthermore, the DNA vaccine prepared using TGEV-S1 and porcine IL-12 could induce excellent humoral and cellular immune responses.
由传染性胃肠炎病毒(TGEV)引起的传染性胃肠炎(TGE)是猪的一种高度传染性疾病。疫苗接种是预防TGEV感染的有效方法。在此,我们在小鼠模型中评估了TGEV S1作为DNA疫苗以及猪白细胞介素(pIL)-12作为佐剂的潜力。构建了一种携带TGEV S1基因的DNA疫苗,以在实验小鼠模型中诱导免疫反应;pIL-12被选作该DNA疫苗中的免疫佐剂。将pVAX1-(TGEV-S1)和pVAX1-(pIL-12)载体转染到BHK-21细胞中并在体外表达。实验小鼠通过肌肉注射途径分别用每种重组质粒和对照进行免疫。分析从血液和脾脏中分离的淋巴细胞的增殖、细胞毒性活性以及CD4和CD8细胞群体。还分析了血清中酶联免疫吸附测定(ELISA)中的TGEV S1滴度、TGEV中和抗体以及干扰素(IFN)-γ和白细胞介素-4的浓度。质粒pVAX1-(TGEV-S1)和pVAX1-(pIL-12)可在BHK-21细胞中表达,并且pVAX1-(TGEV-S1)和pVAX1-(pIL-12)的组合可使所有标志物显著增加。pIL-12可作为TGEV-S1 DNA疫苗中的免疫佐剂。此外,使用TGEV-S1和猪IL-12制备的DNA疫苗可诱导出色的体液和细胞免疫反应。