水痘-带状疱疹病毒在Vero细胞中的传代培养及抗水痘血清的制备

Subculture of varicella-zoster virus in Vero cells and preparation of antiserum

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 subculture varicella-zoster virus（VZV） in Vero cells and prepare the antiserum. Methods VZV 84-2 strain was subcultured in Vero cells, from which antigen was extracted after stable CPE appeared. Rabbits and guinea pigs were immunized with the prepared antigen for 4 times, each at an interval of 14 d. The rabbits were divided into two groups. Adjuvant-free antigen was immunized s.c. to the rabbits in group 1 for the first and the second doses,while the antigen containing complete Freund adjuvant was immunized i.p. to those in the same groups for the third and the forth doses. However, the rabbits in group 2 and the guinea pigs were immunized s.c. with the antigen containing complete Freund adjuvant. The serum samples of animals in various groups were collected 2 weeks after the last immunization for sterility test, determination of neutralizing antibody titer and specific interference test. Results CPE appeared in 80% ~ 90% of Vero cells in which VZV 84-2 strain was subcultured to passage 5, and was stable after passage 10. The protein contents of antigens prepared by using the media containing rabbit and guinea pig sera were 2. 1and 1. 7 mg / ml respectively. The antisera were qualified in sterility. The serum neutralizing antibody titers of rabbits in groups 1 and 2 were 1 ∶ 128 and 1 ∶ 256 respectively, while that of guinea pigs was 1 ∶ 64. The antiserum against VZV showed no interference to the titration of measles（Shanghai 191 strain）, mumps（S79 strain）and rubella（BRDⅡ strain）viruses. Conclusion VZV was subcultured in Vero cells successfully, and antiserum against VZV was prepared, of which the tier was relatively low.