Benzonase酶降解Vero细胞DNA的狂犬病疫苗动物安全性评价

Evaluation of safety of rabies vaccine prepared by degradation of Vero cell DNA with Benzonase in animals

目的对Benzonase非限制性核酸内切酶降解Vero细胞DNA的狂犬病疫苗进行动物安全性评价。方法依据《化学药物刺激性、过敏性和溶血性研究技术指导原则》,对经Benzonase酶处理的冻干人用狂犬病疫苗(Vero细胞)进行豚鼠全身主动过敏性试验、体外溶血性试验、大鼠急性毒性试验和家兔肌肉刺激试验,评价疫苗的安全性。结果狂犬病疫苗豚鼠过敏反应发生率为50%,5 min后恢复正常;体外溶血结果为阴性;大鼠急性毒性试验未见异常;家兔肌肉刺激性试验在注射部位肌肉局部出现了极轻度到轻度的间质炎性细胞浸润、肌肉组织变性坏死及间质纤维组织增生,给药后2周,上述病变程度明显缓解。结论利用Benzonase非限制性核酸内切酶降解Vero细胞DNA的狂犬病疫苗(酶残留量低于0.2 ng/ml)安全性良好,不影响其临床使用。

Objective To evaluate the safety of rabies vaccine prepared by degradation of Vero cell DNA with Benzonase in animals. Methods According to the Technical Guideline for Irritation, Aanphylaxis and Haemalysis of Chemical Drugs, the freeze-dried rabies vaccine （Veto cells） for human use after treatment with Benzonase was subjected to systemic active anaphyla^is test and in vitro hemolysis test in guinea pigs, acute toxicity test in rats and muscle stimulation test in rabbits to evaluate the safety. Results The incidence of anaphylactic reactions of the vaccine in guinea pigs was 50% and the animals with reactions were recovered to normal 5 rain later. The result of hemolysis test was negative. No abnormality was observed in acute toxicity test. Extremely mild or mild interstitial inflammatory cell infiltration, musculature degeneration and necrosis as well as desmoplasia were observed in injection sites of rabbits, of which the degrees were relieved 2 weeks later. Conclusion The rabies vaccine prepared by degradation of Vero cell DNA with Benzonase, with a residual Benzonase content of less than 0. 2 ng/ml, showed high safety and was suitable for clinical application.