SARS冠状病毒灭活疫苗研究初报

目的研制出安全、有效、质量可控的SARS冠状病毒灭活疫苗;同时,制备能用于SARS被动免疫和治疗用的异源高效价特异性抗体,用于SARS的免疫防治.方法采用细胞培养法,从非典型肺炎患者标本中分离获得病毒株,通过电镜、血清学以及RT-PCR技术进行病毒鉴定.运用细胞规模化培养技术大量制备SARS冠状病毒.用温度灭活法和甲醛灭活法对SARS冠状病毒进行灭活处理,筛选灭活条件.将不同类型和不同剂量的佐剂与灭活病毒加以配伍,优化SARS灭活疫苗;同时,以小鼠、兔、猴等为实验对象,进行灭活疫苗的免疫原性评价.用灭活疫苗免疫马,获取异源(马)高免血清,并进行应用效果评价.结果从分离得到的SARS冠状病毒中,优选出适合疫苗制作的SARS病毒株2株-F69和Y3,初步建立了大规模培养SARS病毒的技术体系,病毒TCID50达到106以上.确定了SARS病毒的灭活方法,优选出灭活疫苗的佐剂,并已生产出一定量的SARS灭活疫苗.将灭活疫苗初步用于小鼠、兔等的免疫接种,对其体液免疫和细胞免疫效果进行了初步检测.结论试验研制的SARS灭活疫苗具有较强的免疫原性,设计的灭活疫苗生产技术路线是可行的,进一步的研究工作结束后,可望进行规模化开发生产.

Preiliminary study on inactivated SARS coronavirus vaccine

Objective To develop safe, effective and quality- controllable inactivated vaccine of SARS coronavirus, and prepare high titer specific heleroantibody for use of SARS passive immunity and therapy. Methods SARS coronavirus was isolated from SARS patients by cell culture and identified by electron microscope, immunohematology methods and RT- PCR. Large scale cell culture model was established to get large quantity of virus. SARS coronavirus was treated by thermal inactivation and formalin inactiva-tion, and optimal inactivation factors were determined. The inactivated coronavirus was blended with different dose of defferent adjuvants before immunogenicity assay in mouse and rabbit and the best was selected. High liter specific heteroantibody was prepared by immune horse and was evaluated. Results More than 10 strains from about 100 isolated coronavirus strains were steadily cultured, and F69 and zhonger- Y3 were selected as vaccine slrains. Coronavirus culture with tiler of about 106~108 TCID50 were obtained by the established large- scale culture model. Optimal inactivation methods and the best adjuvant were determined. Inactivated vaccine were prepared and applied to mouse and rabbit, from which respective antibodies were oblained. The results indicated that strong immunogenicity of the inactivated vaccine was remained. Conclusion The prepared vaccine is of high immunogenicity so that the technique for producing inactivated vaccine is feasible.