不同佐剂对猪圆环病毒衣壳蛋白免疫原性的影响

试验旨在研究猪圆环病毒不同佐剂疫苗的制备及其对免疫原性的影响。将去除信号肽的PCV2（porcine circovirus type 2，PCV2）Cap蛋白基因连接在pET-28a载体上，进行诱导表达，用脱氧胆酸钠（DOC）和低浓度尿素对表达产物进行溶解，制备氢氧化铝胶体佐剂、脂质体佐剂、弗氏佐剂、白油佐剂、蜂胶佐剂，并与纯化的PCV2-Cap蛋白混合制成亚单位疫苗，以商品化的灭活疫苗作为阳性对照，免疫小鼠，并使用ELISA方法检测动物血清中抗体及细胞因子含量的变化，评估其免疫保护效果。结果显示，表达产物以包涵体的形式存在，使用DOC溶解包涵体可省略蛋白复性步骤，且纯化方法简单，获得纯度较高的衣壳蛋白；ELISA检测结果表明PCV2-Cap蛋白能诱导产生特异性较高的抗体；水系佐剂制备的亚单位疫苗具有较高的免疫活性，为亚单位疫苗的商品化提供重要的数据支持。

Effects of Different Adjuvants on the Immunogenicity of PCV2-Cap Protein

The study was aimed to prepare vaccines with different adjuvants,and research its effects on immunogenicity.The PCV2 Cap gene with its signal peptide removed was connected to pET-28a vector,and then was induced to express,using sodium deoxycholate（DOC）and low concentration of urea to dissolve the inclusion body.Different adjuvants,such as alum-based adjuvants,liposome adjuvants,propolis adjuvants,white oil adjuvants and Freund adjuvant were prepared,together with protein purified to make up subunit vaccines,and commercial inactivated vaccine as positive control,immuning mice,and ELISA method were used to detect changes in concentrations of animal serum antibody and cytokines,evaluated the immune protective effect.Results showed that the expression product in the form of inclusion body,using the DOC could omit dissolving inclusion body protein renaturation steps,and the purification method was simple,the capsid protein obtained had high purity.ELISA assays showed that PCV2-Cap had good immunogenicity.And we found that the water system adjuvants had high immune activity,this could provide important previous experimental data for the commercialization of the subunit vaccine.