【摘要】 目的　研究SARS冠状病毒棘突蛋白受体结合部位S1的免疫原性 ,为SARS的实验诊断和新型疫苗的研究提供依据。方法　用克隆有哺乳动物细胞密码子优化的SARS CoVS1基因的质粒pcDNA3 1 S1或P S1Ig转染 2 93T细胞 ,用细胞的上清液纯化S1蛋白。以pcDNA3 1 S1质粒对BALB c小鼠进行 2次基因免疫 ,以纯化的S1蛋白进行加强免疫。用ELISA法检测小鼠抗SARS CoV的特异性IgG抗体 ,并在VeroE6细胞上做体外中和实验 ,检测中和抗体。结果　S1蛋白诱导小鼠产生抗SARS CoV的特异性抗体 ;1∶14 99 6 8稀释的S1蛋白免疫的小鼠血清可保护 5 0 %的细胞对 10 0 0TCID50 的病毒攻击 ,而阴性对照血清不能保护细胞对病毒的感染。结论　SARS冠状病毒棘突蛋白受体结合部位S1能有效诱导机体产生具有高效保护作用的中和抗体免疫反应 ,可望发展成为理想的SARS棘突蛋白亚单位疫苗。更多还原

【Abstract】 Objective To study the antigenicity of SARS associated coronavirus (CoV) spike S1 (12-672Aa) domain Methods BALB/c mice were immunized with a plasmid bearing codon-optimized SARS-CoV (Tor2 strain) S1 domain and then boosted with purified S1 protein; the SARS-CoV specific IgG antibody was tested by ELISA and neutralization antibody was determined by in vitro microneutralization assay Results S1 domain of SARS-CoV spike, which has been demonstrated harboring the receptor binding domain, successfully elicited SARS-CoV specific IgG antibody in mouse after combined immunization with DNA and purified S1 protein; the antibody elicited solely by S1 could potently neutralize SARS-CoV (HKU-39849) in vitro, 50% of 1 000 TCID 50 SARS-CoV challenged cells were protected from viral infection by a 1∶1 49968 dilution of mice sera immunized with S1 protein, but negative control sera showed no protection Conclusion S1 domain of SARS-CoV spike protein, which is responsible for receptor binding, can efficiently and sufficiently induce highly potent neutralizing antibody in mice This result suggested that S1 domain could be an effective subunit vaccines against SARS-CoV更多还原