评价肺炎链球菌多糖结合疫苗效力的调理吞噬试验条件的优化  

Optimization of Condition for Opsonophagocytic Assay for Evaluation of Potency of Pneumococcus Polysaccharide Conjugate Vaccine

作　　者：刘倩[1] 陈晓航[1] 任克明[1] 张轶[1] 王玺[1] 沈荣[1] 

机构地区：[1]兰州生物制品研究所第三研究室,兰州730046

出　　处：《中国生物制品学杂志》2010年第11期1248-1253,共6页Chinese Journal of Biologicals

基　　金：2006-2009年甘肃省科技攻关计划项目(2GS063-A43-011)

摘　　要：目的优化评价肺炎链球菌多糖结合疫苗效力的调理吞噬试验(Opsonophagocytic assay,OPA)的条件。方法参考美国阿拉巴马大学的WHO指定实验室于2007年1月发布的A.02版实验规程,优化OPA的试验条件。分别以抗性和非抗性肺炎链球菌菌株作为靶细菌检测其对应型结合物受试血清的调理吞噬效价后,再与相应的ELISA效价比较,分析二者的相关性。结果 0.8%二甲基甲酰胺(DMF)诱导HL-60细胞分化4d后,细胞表面标志表达量分别为:CD11b和CD35≥55%、CD71≤15%,活细胞比例≥65%;各型肺炎链球菌工作种子批的复活率≥80%;补体的非特异性杀伤率为18%;各血清型均表现出良好的杀菌活性,且OPA法与ELISA法测定的血清抗体效价比较,相关系数和斜率非常接近。结论优化的OPA条件符合该试验要求,可用于肺炎链球菌荚膜多糖结合疫苗的效力评价。Objective To optimize the condition for opsonophagocytic assay（OPA）for evaluation of potency of pneumococcus polysaccharide conjugate vaccine.Methods The experimental condition for OPA was optimized according to the protocol for opsonophagocytic assay against streptococcus pneumoniae（version A.02,January 2007）published by WHO reference laboratory in US University of Alabama.The opsonophagocytic titers of drug-resistant and drug-sensitive pneumococcus strains against the sera of mice immunized with pneumococcus polysaccharide conjugates of the corresponding types were determined and compared with the corresponding ELISA titers,between which the relationship was analyzed.Results After HL-60 cells were differentiated for 4 d under induction of 0.8% DMF,both the expression of cell surface markers CD11b and CD35 were not less than 55%,and that of CD71 was not more than 15%,and the proportion of live cells was not less than 65%.The recovery rate of working seed lot of pneumococcus of various types were not more than 80%,while the non-specific killing rate of complement was 18%.All the serotypes showed high bactericidal activity,and similar correlation coefficients and slopes were observed between OPA and ELISA titers.Conclusions The optimized condition met the requirements for OPA and might be used for the evaluation of potency of pneumococcus polysaccharide conjugate vaccine.

关 键 词：肺炎链球菌 疫苗 结合 效力 调理吞噬试验 

分 类 号：R378.14[医药卫生—病原生物学] R392.33[医药卫生—基础医学]