不同裂解工艺制备的甲型H1N1流感病毒裂解疫苗的透射电子显微镜观察  

Transmission electron microscopy of influenza A H1N1 virus split vaccines prepared by various cleavage procedures

作　　者：李楠[1] 王亚军[2] 张雪梅[2] 郝镯[1] 温志英 张赐强[2] 李树民[1] 李晓波[2] 李吉平[1] 

机构地区：[1]中国人民解放军军事医学科学院军事兽医研究所吉林省人兽共患病预防与控制重点实验室,吉林长春130122 [2]长春生物制品研究所有限责任公司,吉林长春130062

出　　处：《中国生物制品学杂志》2014年第3期296-298,304,共4页Chinese Journal of Biologicals

摘　　要：目的透射电子显微镜观察不同工艺制备的甲型H1N1流感病毒裂解疫苗形态，筛选最佳裂解工艺，为疫苗的大规模生产提供参考依据。方法应用透射电镜对4种不同裂解工艺制备的甲型H1N1流感病毒疫苗进行形态观察，采用中和试验分析其免疫原性，异常毒性试验分析其安全性。结果透射电镜观察显示4种样品均无完整病毒，样品1裂解程度最大，囊膜破裂，多数为病毒碎片；样品2大多数囊膜未破损，缺少纤突；样品3较完整的病毒颗粒略小（约30-40nm），纤突清晰；样品4大多数病毒颗粒保留着纤突，但纤突已不整齐清晰。样品2组小鼠中和抗体滴度最低，而样品1、3、4组较高，与样品2组相比，差异明显，其中以样品3组最高；样品2组小鼠、豚鼠体重增加不明显，而样品1、3、4组小鼠、豚鼠体重增加明显，与样品2组相比差异显著，其中以样品3组最明显。结论电镜观察可直观反映疫苗裂解程度；3种裂解工艺均可用于疫苗生产，其中样品3具有较好的免疫原性和安全性，其工艺可作为最佳裂解工艺用于后续疫苗生产。Objective To observe the morphological characteristics of influenza A H1N1 virus split vaccine by transmission electron microscopy, optimize the procedure for cleavage and provide a basis for large scale production of the vaccine. Methods The influenza A H1N1 virus split vaccines prepared by four cleavage procedures were observed by transmission electron microscopy, of which the immunogenicity was analyzed by neutralization test, while the safety by abnormal toxicity test. Results No intact virus particles were observed in all the vaccines prepared by four cleavage procedures. Sample 1 showed the highest split leveL, which were virus debris due to breakage of membrane. Most of membrane of sample 2 showed no breakage, and no spike was observed. The virus particles in sample 3 were slightly smaller than intact virus particles, which were at sizes of about 30 - 40 nm, and clear spikes were observed. However, irregular and unclear spikes were observed in most virus particles in sample 4. The neutralizing antibody titers of mice immunized with sample 2 was the lowest, while those in the other three groups were significantly higher, of which that of mice immunized with sample 3 was the highest. The bodyweights of mice and guinea pigs immunized with sample 2 showed no significant increase, while those in the other three groups increased significantly, of which that of animals immunized with sample 3 showed the most significant increase. Conclusion Electron microscopy reflected the cleavage level of vaccine directly. Three procedures may be used for vaccine production. The vaccine prepared by procedure 3 showed high immunogenicity and safety, indicating that the procedure may be used as optimal cleavage procedure for subsequent vaccine production.

关 键 词：流感病毒A型 H1N1亚型 疫苗 裂解工艺 透射电子显微镜 

分 类 号：R373.13[医药卫生—病原生物学] R392-33[医药卫生—基础医学]