SARS-CoV基因疫苗的构建及免疫原性初步研究

Development of SARS-CoV vaccines and evaluation of their immunogenicity

目的构建SARS病毒E、M、N基因疫苗,研究其免疫原性。方法通过PCR扩增获得SARS-CoVE、M、N基因片断,将其分别克隆入真核表达载体pVAX1,将所得真核表达质粒pVAX1-E、pVAX1-M和pVAX1-N分别肌肉注射小鼠和兔子,用ELISA法检测血清抗体,观察实验期间动物的体重变化,处死后对重要脏器进行病理检查。结果pVAX1-E、pVAX1-M和pVAX1-N基因测序结果与基因库中公布的一致。3种基因疫苗肌肉注射免疫小鼠和兔子后均能诱导产生特异性抗SARS-CoV抗体;动物一般状况和体重变化与对照组差异无显著性;重要脏器无明显病理改变。结论pVAX1-E、pVAX1-M和pVAX1-N疫苗能够诱导机体产生特异的体液免疫应答,具有较好的免疫原性和安全性,为SARS疫苗的研制奠定了良好基础。

[Objective] To develop vaccines of envelop gene（E）, membrane gene（M） and nucleocapsid（N） gene from SARS-CoV, and to evaluate their immunogenicity. [Method] E, M and N genes were obtained by PCR, and then cloned into eukaryotic expression vector pVAX1. Mice and rabbits were immunized by intramuscular injection with the recombinant plasmids. The antigen-specific antibodies were determined by ELISA. The weight change of the animals were observed. Their organs were taken, and pathological changes were observed. [Results] The sequences of recombinant plasmids were identical to those in the Genebank. Of the animals immunized with pVAX1-E, pVAX1-M and pVAX1-N DNA vaccines, the level of specific antibodies against SARS-CoV increased significantly in sera. The weight change and pathological result of the animal immunized with DNA vaccines showed slight difference with negative control. [Conclusion] The pVAX1-E, pVAX1-M and pVAX1-N DNA vaccines can induce specific humoral immune responses in the mice and rabbits. they＇ve shown good immunogenicity and safety. They could be used as candidate vaccines against SARS.