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(0.01<P<0.05),且X4550/pVAX1-P32免疫组能诱导持续时间更长的抗体水平(P<0.01),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。
A novel oral live vector DNA vaccine encoding a P32 of goatpox virus without the transmembrane and hydrophobic domains,was constructed by using recombinant attenuated Salmonella typhimurium X4550.Its safety,stability,immunogenicity and immune effects were detected and evaluated comprehensively by continuous cultivation in vitro,megadose inoculation test,immunohistochemical test, ELISA, etc.The results showed that X4550/pVAX1-P32 was of safety and stability in vitro and in vivo for BALB/c mice,and the survival rate reached to 100% after oral immunization with 1× 1010 CFUX4550/pVAX1-P32.The X4550/pVAX1-P32 vaccine could be passaged stably above 100 generations and recombinant S.typhimurium was isolated from the spleen 10 days post-immunization.The attenuated S.typhimurium X4550 could delivery DNA to immunologic effector cells effectively and expressbioactive P32 protein.Tawny particles were detected in the spleen cells from the immunized mice by immunohistochemistry test.The results of ELISA showed that X4550/pVAX1-P32 induced a longer persistent period（P〈0.01） and higher antibody level than pVAX1-P32（0.01〈P〈0.05）.These results indicate that the live vector DNA vaccine has good immunogenicity and can induce a lasting and high-level immune response, providing a basis for development of novel vaccines against goatpox.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2017年第3期269-278,共10页
Chinese Veterinary Science
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)
