CHO细胞簇集试验在无细胞百日咳疫苗质量控制中的应用

Application of Chinese hamster ovary cell clustering assay in quality control of acellular pertussis vaccine

目的探讨CHO细胞簇集试验在无细胞百日咳疫苗质量控制中的应用。方法采用CHO细胞簇集试验比较分析不同来源的CHO细胞对百日咳毒素（pertussis toxin,PT）簇集活性。将豚鼠随机分为进口疫苗Lot 1~3实验组、国产疫苗Lot 4实验组、参考疫苗对照组,同时设未免疫对照组。分别于第0和21天,经豚鼠腹部皮下注射0.5 ml疫苗。于初免后第28天,经心脏采血,分离血清,采用ELISA法检测PT抗体滴度,CHO细胞簇集试验检测中和抗体滴度。结果 PT对不同来源的CHO细胞簇集能力存在一定差异,其中以来自ATCC的CHO细胞的簇集最敏感。各无细胞百日咳疫苗在豚鼠动物模型上均能诱导较好的PT抗体免疫反应,进口疫苗与参考疫苗及国产疫苗相比,差异均无统计学意义（P〉0.05）;两种方法的检测结果具有一定的相关性,Spearman相关系数为0.474。结论证实了CHO细胞簇集试验用于百日咳疫苗纯化过程中质量控制和PT免疫原性分析的适用性,为加强和完善我国无细胞百日咳疫苗的质量控制奠定了基础。

Objective To investigate the application of Chinese hamster ovary（CHO）cell clustering assay in quality control of acellular pertussis vaccine. Methods The clustering activities of CHO cells from different origins induced by pertussis toxin（PT） were compared by CHO cell clustering test. Guinea pigs were randomly divided into Lot 1 ~ 3（imported vaccines）,Lot 4（domestic vaccine）and reference vaccine groups,using those unimmunized as control. The guinea pigs were immunized with 0. 5 ml of vaccine on day 0 and 21,of which the serum samples were collected on day 28 and determined for PT antibody titer by ELISA,and for neutralizing antibody titer by CHO cell clustering assay. Results The clustering abilities of PT to CHO cells from various origins showed a certain difference,of which that to the cells from ATCC was the highest. Various acellular pertussis vaccines induced high antibody titers against PT in guinea pigs. The antibody titers induced by imported vaccines showed no significant differences with those by reference and domestic vaccines（P〉 0. 05）. The determination results by ELISA and by CHO cell clustering assay showed a certain correlation,with a Spearman correlation coefficient of 0. 474. Conclusion The study demonstrated that CHO cell clustering assay was suitable for the quality control during purification of pertussis vaccine and analysis of immunogenicity of PT,which laid a foundation of strengthening and improvement of quality control of acellular pertussis vaccine.