山羊痘病毒P32抗原基因口服活载体DNA疫苗的构建

发布时间：2018-05-08 19:03

本文选题：山羊痘病毒 + P基因　；参考：《中国兽医科学》2017年03期

【摘要】：以减毒鼠伤寒沙门菌X 4550为载体,构建编码缺失跨膜区和疏水区的山羊痘病毒P32基因的新型口服活载体D N A疫苗,通过体外传代培养、大剂量接种试验、免疫组织化学试验、ELISA等对其安全性、稳定性、免疫原性及免疫效果等进行综合检测评价。结果表明,X4550/pVAX1-P32在体外可稳定传代100代以上,小鼠口服1×10~(10)CF U X4550/pVAX1-P32后存活率为100%,接种后第10天仍能从脾中分离到X4550/pVAX1-P32,表明X4550/pVAX1-P32在体内外均具有良好的安全性和稳定性。免疫组织化学试验结果显示免疫小鼠脾细胞内有黄褐色粗颗粒,表明X4550能将pVAX1-P32有效递呈至免疫效应细胞并在其内表达具有生物活性的P32蛋白。ELISA结果显示,X4550/pVAX1-P32免疫组的抗体水平显著高于pVAX1-P32免疫组(0.01P0.05),且X4550/pVAX1-P32免疫组能诱导持续时间更长的抗体水平(P0.01),表明活载体疫苗X4550/pVAX1-P32具有良好的免疫原性,能够诱导机体产生持久的、高水平的免疫应答,这为羊痘新型疫苗的研究和应用奠定了基础。
[Abstract]:Using attenuated Salmonella typhimurium X 4550 as vector, a novel oral live vector D N A vaccine encoding goat pox virus P32 gene with missing transmembrane and hydrophobic regions was constructed. The safety, stability, immunogenicity and immune effect were evaluated by Elisa. The results showed that X4550 / pVAX1-P32 could be passed through more than 100 passages in vitro, and the survival rate of mice was 100 after taking 1 脳 10~(10)CF U X4550/pVAX1-P32. X4550 / pVAX1-P32 could still be isolated from the spleen on the 10th day after inoculation, indicating that X4550/pVAX1-P32 had good safety and stability in vivo and in vitro. Immunohistochemical test showed that the spleen cells of immunized mice had yellow brown coarse granules. The results showed that X4550 could effectively present pVAX1-P32 to immune effector cells and express biologically active P32 protein. Elisa results showed that the antibody level of X4550 / pVAX1-P32 immunized group was significantly higher than that of pVAX1-P32 immunized group (0.01P0.05N), and X4550/pVAX1-P32 immunized group could induce longer duration. The antibody level of P0.01a indicated that the live vector vaccine X4550/pVAX1-P32 had good immunogenicity. It can induce persistent and high level immune response, which lays the foundation for the research and application of new vaccine of sheep pox.
【作者单位】：新疆大学生命科学与技术学院;中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室;
【基金】：新疆维吾尔自治区科技支疆项目(210-60879) 新疆维吾尔自治区高层次引进人才项目(1101/45051)
【分类号】：S852.4

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