狂犬病病毒糖蛋白的犬2型腺病毒活载体重组疫苗安全评价  

Biosafety Evaluation of Canine Adenovirus type-2 Expressing Rabies Virus Glycoprotein in Dogs

作　　者：韩乃君[1] 刘晔[2] 扈荣良[2] 张乐萃[1] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109 [2]军事医学科学院兽医研究所

出　　处：《青岛农业大学学报（自然科学版）》2008年第3期236-241,共6页Journal of Qingdao Agricultural University(Natural Science)

摘　　要：本试验用狂犬病病毒糖蛋白-犬2型腺病毒重组疫苗口服或注射免疫500条受试犬,免疫一次,采样测定其遗传稳定性、在免疫犬体内的分布、存留时间、排泄规律、高剂量接种对免疫犬的影响,检测疫苗生产人员、试验人员以及试验区周围动物体内犬腺病毒特异性抗体以及免疫犬体内诱导产生的抗体水平和消长规律,监测试验区域外源基因存留情况。通过以上方法来评价重组疫苗环境释放阶段的安全性。结果表明,重组疫苗体外传代79次、体内传代11次后,核酸序列和生物特性未发生改变。重组病毒在免疫犬大肠和小肠短暂存留,在犬粪便中短期存在。高倍剂量免疫受试犬无急性、亚急性和慢性毒性作用,无致畸作用,子代犬均发育正常。重组疫苗生产人员和试验人员体内未检测出犬腺病毒特异性抗体;检测试验区周围除个别犬由于自然感染犬腺病毒野毒株而带有低水平的犬腺病毒抗体(但无狂犬病抗体),其余动物均无犬腺病毒抗体。诱导的抗体平均水平在国际最低保护水平(0.5IU/ml)以上,抗体持续时间至少为54周。试验表明重组疫苗以口服和注射两种途径免疫,均具有较高的安全性和较好的免疫效果。To evaluate the biosafety in the enviroment release of the recombinant canine adenovirus type -2 expressing the rabies virus glycoprotein, 500 unvaccinated dogs were split into two groups, 250 in each group. Group 1 was intramuscularly injected with 1 ml （ 107.75 TCID50 /ml） and Group 2 was orally inoculated with 3 ml （ 107.75 TCID50/ml）, respectively. At the vaccination and after, the dog sera were collected weekly or biweekly and the following items were evaluated ： the genetic stability of the recombinant vaccine in cell passage and in dogs ; the existence and sustaining of the recombinant virus in the body and in the feces of the immunized dogs; the toxic effect of the recombinant virus on dogs immunized with overdoes; the sepcific antibody against the recombinant virus in the bodies of producers, the experimenters and animals around the experimental area; the contamination of the exogenous （ i. e. , the glycoprotein） gene in the environment around the experimental area during the experiment and 6 months after the experiment; the neutralizing antibody level in dogs and the kinetics of the neutralizing antibody. Results showed that after 79 passages in MDCK cells and 11 passages in dogs, no deletion or recombination of the foreign gene in the recombinant virus was detected. And no specific nucleotide sequence of the recombinant virus was assayed in the bacteria isolated from the tissues of the immunized dogs. No contamination of the recombinant virus and the glycoprotein gene was found out in the environment. Increasing the amount of the recombi- nant virus did no harm to the dogs and no toxicity was observed. No specific antibody against the recombinant virus was assayed in the producers, experimenters and the animals around the experimental area except for several dogs naturally infected with the canine adenovirus. Neutralizing antibody was detectable monthly after the intramuscular and oral vaccination, respectively. The antibody could persist for 54 weeks with an effective level of protection

关 键 词：狂犬病 重组疫苗 环境释放 安全性评价 免疫研究 

分 类 号：S852.65[农业科学—基础兽医学]