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目的在Vero细胞中传代培养水痘-带状疱疹病毒(varicella-herpes zoster virus,VZV),并制备抗水痘血清。方法将VZV(北京株VZV84-2)在Vero细胞中传代培养,待细胞病变(CPE)稳定后制备抗原,分别免疫家兔(分为1组和2组)和豚鼠(仅1组),均免疫4次,每次间隔14 d,其中除家兔1组第3、4次免疫经耳静脉注射不含佐剂的免疫原外,其他均经背部皮下免疫含弗氏完全佐剂的免疫原。于末次免疫后2周经颈动脉采血,分离血清,进行无菌试验、中和抗体效价检测及特异性干扰试验。结果 VZV(北京株84-2)在Vero细胞中传至第5代时,CPE可达80%~90%,传至第10代时,CPE稳定。用含兔及豚鼠血清培养基制备的免疫原的蛋白含量分别为2.1和1.7 mg/ml。抗血清无菌检测合格;家兔1组、豚鼠组、家兔2组的血清中和抗体效价分别为1∶128、1∶256、1∶64;抗水痘血清对麻疹(沪191株)、腮腺炎(S79株)、风疹(BRDⅡ株)疫苗的滴定无干扰。结论成功将VZV在Vero细胞中进行传代培养,并制备了抗水痘血清,但效价较低,有待进一步提高。
Objective To subcultivate varicella-herpes zoster virus (VZV) in Vero cells and prepare anti-varicella serum. Methods VZV (Beijing strain VZV84-2) was subcultured in Vero cells. After stabilization of cytopathic effect (CPE), antigens were prepared and immunized rabbits (divided into groups 1 and 2) and guinea pigs Immunization 4 times, each interval of 14 d, except rabbits in group 1 and group 3 were immunized via the orthotopic intravenous injection of adjuvant-free immunogens, the other were subcutaneously immunized with Freund’s complete adjuvant immunogen . Serum was collected from the carotid artery 2 weeks after the last immunization and the serum was separated for sterility test, neutralizing antibody titer and specific interference test. Results VZV (Beijing strain 84-2) reached the fifth generation in Vero cells, and the CPE reached 80% -90%. The CPE was stable at passage 10. The protein content of the immunogen prepared with rabbit and guinea pig serum medium was 2.1 and 1.7 mg / ml, respectively. Serum neutralizing antibody titers in rabbits 1 group, guinea pig group 2 and rabbits 2 were respectively 1:128, 1:256, 1:64; Anti-chickenpox serum against measles (Shanghai strain 191) , Mumps (S79), rubella (BRDII strain) titration of the vaccine without interference. Conclusion VZV was successfully subcultured in Vero cells and anti-chickenpox serum was prepared, but its titer was low and needs to be further improved.