甲型H1N1流感病毒裂解疫苗动物体内的安全性和免疫原性

Safety and immunogenicity of influenza A H1N1 virus split vaccine in animals

目的 评价甲型H1N1流感病毒裂解疫苗在动物体内的安全性和免疫原性。方法 按《中国药典》三部（2005版）方法进行异常毒性试验。将小鼠随机分为7组,每组10只,实验组小鼠分别经小鼠后腿胫前肌肌肉注射不同血凝素含量（15、30、45μg/0.5 ml）的甲型H1N1流感病毒裂解疫苗,同时设疫苗对照组（相应规格的季节性H1N1流感病毒裂解疫苗）及阴性对照组（PBS）,注射剂量均为0.1 ml/只,间隔21 d加强免疫1次,分别于初免后0、21、28、35及42 d采集眼静脉血,分离血清,检测血凝抑制（hemagglutination inhibition,HI）抗体水平。结果 异常毒性试验结果符合《中国药典》三部（2005版）判定标准。与阴性对照组比较,初免后21、28、35、42 d,各剂量实验组小鼠血清抗体水平均明显升高,21 d即上升到较高水平,28 d达到峰值（P均〈0.01）;初免后0、21、28、35、42 d,除35 d外,均高于相应剂量疫苗对照组（P均〉0.05）。初免后21 d,15、30、45μg/0.5 ml剂量实验组小鼠血清中HI抗体阳转率分别为80%、90%和90%,HI抗体保护率分别为100%、90%和90%,与相应剂量疫苗对照组相比,差异均无统计学意义（P均〉0.05）;初免后28、35、42 d,15、45μg/0.5 ml剂量实验组HI抗体阳转率维持在80%以上,30μg/0.5 ml剂量实验组HI抗体阳转率略低,而各剂量实验组HI抗体保护率均在90%～100%之间。结论 甲型H1N1流感病毒裂解疫苗在昆明小鼠中具有良好的安全性和免疫原性。

Objective To evaluate the safety and immunogenicity of influenza A H1N1 virus split vaccine in animals. Methods Abnormal toxicity test was performed according to the requirements in Chinese Pharmacopoeia （Volume Ⅲ, 2005 edition）. Kunming mice were randomly divided into seven groups, ten for each. The mice in three test groups were injected i. m. with influenza A H1N1 virus split vaccine at hemagglutinin （HA） contents of 15, 30 and 45 μg/0. 5 ml, while those in three parallel control groups with seasonal influenza H1N1 virus split vaccine at the corresponding HA contents, and those in negative control group with PBS, respectively. The vaccines were given at a dosage of 0. 1 ml for 2 times at an interval of 21 d. Venous blood samples were collected 0, 21, 28, 35 and 42 d after the first immunization, from which sera were separated and determined for hemagglutination inhibition （HI） antibody level. Results The abnormal toxicity test result met the requirements in Chinese Pharmacopoeia （Volume HI, 2005 edition）. The serum antibody levels in three test groups 21, 28, 35 and 42 d after the first immunization increased significantly as compared with those in negative control group, which reached a high level on 21 d and a peak value on 28 d （each P 〈 0. 01）. However, the antibody levels in test groups 0, 21, 28 and 42 d were higher than those immunized with the control vaccine at the cor-responding HA contents （P 〉 0. 05）. The serum HI antibody positive rates of mice 21 d after immunization with test vac- cine at HA contents of 15, 30 and 45 μg/0. 5 ml were 80%, 90% and 90%, while the protection rates were 100%, 90% and 90%, respectively, which showed no significant difference with those in parallel control groups （each P 〉 0. 05 ）. The HI antibody positive rates of mice 28, 35 and 42 d after immunization with 15 and 45 μg/0. 5 ml were more than 80%, while that with 30 μg/0. 5 ml was slightly low. However, the protection rates of HI antibody in three test groups were 90% - 100%. Conclusion Influenza A H1N1 virus split vaccine showed good safety and immunogenicity in Kunming mice.