评价肺炎链球菌疫苗免疫效果的功能性抗体检测方法的建立  

Establishment of functional antibody detection method for evaluation of potency of pneumococcal vaccine

作　　者：李江姣[1] 杜慧竟[1] 石继春[1] 徐苗[1] 叶强[1] 

机构地区：[1]中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《中国医药导报》2014年第20期4-8,共5页China Medical Herald

基　　金：国家高技术研究发展计划(863计划)项目(编号2012AA02A402);中国食品药品检定研究院中青年发展研究基金(编号2012B15)

摘　　要：目的建立肺炎球菌疫苗免疫效果评价的功能性抗体滴度检测方法。方法参考美国阿拉巴马大学的多重调理吞噬实验(MOPA)方法,构建并鉴定HL-60效应细胞库、肺炎球菌工作菌种库和补体;在此基础上对5份血清样本进行调理吞噬活性检测。结果 HL-60细胞分化后表面标志表达量CD35为71.78%,CD71为10.97%,活细胞比例为72.96%。13个血清型的肺炎链球菌工作菌种的冻存复活率均高于90%;抗生素抗性与敏感性与预期结果一致;工作稀释度均≥1000。补体的非特异性杀菌率均≤70%。血清样本的杀菌曲线均为标准的S型,平行孔间的差异不超过3倍。结论效应细胞、工作菌种、补体及血清样本的检测结果均符合阿拉巴马大学参考方法的标准要求,成功建立了基于MOPA的功能性抗体检测方法。Objective To establish the functional antibody detection method for evaluation of immune effect of pneumococcus vaccine. Methods The effector cell cultures, working bacteria stocks and complement, which were important components of multiplexed opsonophagocytic killing assay（MOPA） method were constructed and detected for applicability according to the reference method published by WHO reference laboratory in US University of Alabama at Birmingham（UAB）. The opsonophagocytic titers of five sera samples were determined using the established MOPA method.Results After differentiation of HL-60 cells, the expression of cell surface markers CD35 and CD71 were 71.78% and10.97%, respectively, and the proportion of live cells was 72.96%. The percentage recovery of working bacteria stocks of all serotypes were more than 90%. The antibiotic sensitivity and resistance of the frozen assay stocks were accordance with the expected results. The optimal dilution factor for all the assay stocks were more than 1000. The nonspecific killing rates of complement for all serotypes were less than 70%. The sample＇s killing curves for all serotypes were sigmoid shape, and the opsonophagocytic titers of the duplicates were differ by less than 3-fold. Conclusion The detection results of the effector cell cultures, working bacteria stocks, complement and sera samples meet the requirements of reference method of UAB, indicating the MOPA method is successfully established in this study.

关 键 词：肺炎链球菌 疫苗 功能抗体 调理吞噬试验 

分 类 号：R392[医药卫生—免疫学]