作　　者： ; ; ; ; ; ; ; (赵武）;

机构地区： 华中农业大学动物医学院

出　　处： 《生物工程学报》 2005年第5期725-730,共6页

摘　　要： 为了提高表达GP5的猪繁殖与呼吸综合征病毒(PRRSV)DNA疫苗的免疫效应,将具有蛋白转导功能的牛疱疹病毒1型(BHV1)VP22基因插入到经过修饰具有更好免疫原性的PRRSV修饰型ORF5基因(ORF5M)上游,构建VP22和ORF5M融合表达的真核表达质粒pCIVP22ORF5M。经间接免疫荧光试验(IFA)和Westernblot检测证实体外表达后,免疫BALBc小鼠,检测小鼠免疫后的GP5特异性ELISA抗体、抗PRRSV中和抗体和脾淋巴细胞增殖反应,并与非融合的真核表达质粒pCIORF5M进行比较。结果显示,融合表达VP22GP5的DNA疫苗pCIVP22ORF5M诱导的体液免疫和细胞免疫反应均明显高于非融合表达的DNA疫苗pCIORF5M,表明蛋白转导相关蛋白BHV1VP22能显著增强表达GP5的PRRSVDNA疫苗的免疫效应,有效发挥了基因免疫佐剂效应;这为研制PRRSV高效DNA疫苗奠定了基础,同时也为其它疾病的高效新型疫苗研究提供了思路。 To enhance the immuogenicity of DNA vaccines expressing the GP5 protein of Porcine Reproductive and Respiratory Syndrome Virus （PRRSV）, the tegument protein VP22 （encoded by VP22 gene） of Bovine Herpesvirus 1 （BHV-1）,which has been demonstrated to exhibit the unusual protein transduction property, was fused to N-terminus of GP5 of DNA vaccine construct pCI-ORF5M, resulting in pCI-VP22-ORF5M expressing VP22-GP5 fusion protein. The expression of VP22-GP5 fusion protein was confirmed by both indirect immunofluorescence assay （IFA） and Western blot. To investigate its immunogenicity, BALB/c mice were immunized with the fusion expression plasmid pCI-VP22-ORFSM and non-fusion expression plasmid pCI-ORFSM,respectively. The GPS-specific ELISA antibodies, neutralizing antibodies and lymphocyte proliferative responses were evaluated at various time points after primary immunization. The results showed that GPS-specific ELISA antibodies, neutralizing antibodies,and lymphocyte proliferative responses induced by DNA vaccine pCI-VP22-ORFSM were higher significantly than those of DNA vaccine pCI-ORFSM,indicating that fusion expression with BHV-1 VP22 significantly enhances the immuogenicity of DNA vaccine expressing the PRRSV GP5 protein, and that this strategy may also be useful to develop more efficient DNA vaccines against other pathogens.

关 键 词： 牛疱疹病毒 型 猪繁殖与呼吸综合征病毒 基因 蛋白转导 疫苗

领　　域： [农业科学] [农业科学] [农业科学]