微载体培养MEK和Vero细胞试制甲肝灭活疫苗  

Preparation of inactivated hepatitis A vaccine with MEK and Vero cells cultured on microcarriers

作　　者：井申荣[1] 尹卫东[2] 万宗举[3] 李成明[3] 吴炳元[1] 姜述德[1] 

机构地区：[1]中国医学科学院医学生物学研究所,云南昆明650118 [2]唐山怡安生物工程公司,河北唐山063000 [3]中国药品生物制品检定所,北京100050

出　　处：《免疫学杂志》2000年第4期257-260,共4页Immunological Journal

摘　　要：目的探索微载体培养细胞大量制备甲肝病毒抗原及其灭活疫苗的可行性。方法使用 Cytodex- 1培养恒河猴胚肾细胞和 Vero细胞制备 HAV ,经过初步纯化、甲醛灭活、吸附佐剂 ,制成甲肝灭活疫苗 ,免疫昆明种小白鼠 ,测定免疫原性。结果 HAV X株和 W株抗原滴度分别为 1∶ 2 5 6、1∶ 12 8,感染滴度 (log TCID5 0 / m l)分别为 8.5 0、8.17,与静止培养获得的滴度相当。小鼠抗 HAV抗体第 45 d达到峰值 ,滴度分别为 1∶ (96 .0± 78.4)、1∶ (12 8.0± 70 .1)。结论实验性甲肝灭活疫苗具有良好的免疫原性 ,应用微载体培养细胞制备甲肝灭活疫苗是可行的。ObjectiveTo investigate the feasibility of producing HAV and inactivated vaccine with cells cultured on microcarriers.Methods Macaca mulatta embryo kidney(MEK) and Vero cells cultured on Cytodex 1 grew confluent and were inoculated with HAV strains.The harvested virus was purified and inactivated by formaldehyde.Mice were immunized with two strains of experimental inactivated vaccines adsorbed to adjuvant.The immune response was detected by determination of antibody against HAV with ELISA method.Results Four weeks post inoculation with HAV strain X on MEK cells,strain W on Vero cells,HAV antigenic titers reached to 1∶256 in MEK cells and 1∶128 in Vero cells,and infective titers (logTCID 50 /ml) were 8.50 and 8.17,respectively.The HAV titers were almost the same as that obtained from stationary culture.The anti HAV titers after 45 days in immunizing mice with inactivated HAV vaccine strains X and W were up to 1∶ (96.0 ±78.4) and 1∶(128.0±70 1),respectively.ConclusionThe experimental inactivated HAV vaccine has high immunogeni city. It is feasible to produce HAV antigens and inactivated HAV vaccine with microcarrier culture techniques. [

关 键 词：微载体 MEK细胞 VERO细胞 灭活疫苗 甲型肝炎 

分 类 号：R392-33[医药卫生—免疫学]