State Key Laboratory of Animal Nutrition, Beijing Institute of Animal Science and Veterinary Medicine, Chinese Academy of Agricultural Sciences, Beijing 100193, China; College of Animal Science and Technology, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China.
J Virol Methods. 2013 Nov;193(2):525-30. doi: 10.1016/j.jviromet.2013.07.031. Epub 2013 Jul 27.
Peste des petits ruminants (PPR), an acute and highly contagious disease, affects sheep, goats, and some small ruminants. The hemagglutinin (H) glycoprotein of the PPR virus (PPRV) is considered important for inducing protective immune responses. In this study, a suicidal DNA vaccine based on the Semliki Forest virus (SFV) replicon was constructed and tested for its ability to induce immunogenicity in a mouse model. For this, the H gene of PPRV was cloned and inserted into pSCA1, an SFV replicon vector. The resultant plasmid named pSCA1-H was then transfected into BHK-21 cells following which the antigenicity of the expressed protein was confirmed by Western blotting and immunofluorescence. The pSCA1-H plasmid was then injected intramuscularly into BALB/c mice thrice at 2-week intervals. To evaluate the immunogenicity of pSCA1-H, specific antibodies and neutralizing antibodies against PPRV-H were measured using an indirect enzyme-linked immunosorbent assay and a microneutralization test, respectively. Cell-mediated immune responses were also examined using a lymphocyte proliferation assay. The results showed that pSCA1-H could express the H protein in BHK-21 cells. Specific antibodies, neutralizing antibodies, and lymphocyte proliferation responses were all induced in mice. Thus, this suicidal DNA vaccine could be a promising new approach for vaccine development against PPR.
小反刍兽疫(PPR)是一种急性、高传染性疾病,影响绵羊、山羊和一些小反刍动物。PPRV 的血凝素(H)糖蛋白被认为对诱导保护性免疫反应很重要。在这项研究中,构建了一种基于 Semliki Forest 病毒(SFV)复制子的自杀 DNA 疫苗,并在小鼠模型中测试其诱导免疫原性的能力。为此,克隆了 PPRV 的 H 基因并插入到 SFV 复制子载体 pSCA1 中。将得到的质粒命名为 pSCA1-H,然后转染 BHK-21 细胞,通过 Western blot 和免疫荧光确认表达蛋白的抗原性。然后将 pSCA1-H 质粒每隔 2 周肌肉内注射到 BALB/c 小鼠 3 次。为了评估 pSCA1-H 的免疫原性,使用间接酶联免疫吸附试验和微量中和试验分别测量针对 PPRV-H 的特异性抗体和中和抗体。还使用淋巴细胞增殖试验检查了细胞介导的免疫反应。结果表明,pSCA1-H 可以在 BHK-21 细胞中表达 H 蛋白。在小鼠中诱导了特异性抗体、中和抗体和淋巴细胞增殖反应。因此,这种自杀 DNA 疫苗可能是针对 PPR 的疫苗开发的一种有前途的新方法。