流行性出血热病毒传代细胞双价纯化疫苗研究  

Advances on development of purified bivalent vaccine against Epidemic Haemorrhagic Fever (HFRS) prepared on VERO cells

作　　者：杭长寿[1] 解燕乡[1] 王世文[1] 霍子威[1] 张全福[1] 

机构地区：[1]中国预防医学科学院病毒学研究所,100052

出　　处：《医学研究通讯》2001年第5期2-4,共3页Bulletin of Medical Research

基　　金：生物技术领域"九五"国家医学技术攻关专题研究内容

摘　　要：目的我国的流行性出血热主要由汉滩型(HTN)和汉城型(SEO)病毒引起,疫苗的免疫接种是控制和预防本病发生的关键之举。现有的疫苗虽有较好的近期防病效果,但副反应多、抗体水平低,而且由于需要大量的动物种群以提供原代细胞的来源,造成环境污染而疫苗质量又难以控制。本专题的主要研究内容和目的就是用Vero细胞作为生产疫苗的基质,同时用现代较先进的浓缩纯化技术和检测手段,研制成精制双价疫苗。方法两型病毒分别在Vero细胞传代适应至高滴度,大量培养收获上清作为疫苗原液,采用超滤技术浓缩50～100倍,然后通过Sepharose4 FF柱纯化。认真测定抗原和蛋白含量、牛血清含量、细胞DNA含量及必要的试验,精心配制成Vero双价疫苗。结果按我国“生物制品规程”和出血热灭活疫苗临床试验最低标准进行全面系统鉴定,完全符合质量要求。正待批准进行人体临床试验。结论本品有可能作为现有疫苗的替代品。Objective Haemorrhagie Fever with Renal Syndrome (HFRS) in China is mainly caused by two serotypes (or genotypes) of Hantavirus: Hantaan virus and Seoul vims. Vaccination is the key method for the disease control and prevention . The existing vaccines have relatively good short - term prevention effect, but simultaneously with many side - effects and lower antibody titre response. For the use of large amount of animal species to prepare preliminary cell cultures, the environment may be polluted and the quality of vaccines is difficult to guarantee. The objective of the project is to develop purified bivalent vaccine by using Vero eell lines as the basic substitutes, and with more advanced techniques to concentrate, purify and detect. Method Two sero - types of viruses were adapted and passaged on Vero cells to reach their high titres respectively. The supernatant was harvested as the original fluid vaccines.The original fluid vaccines were concentrated 50 to 100 folds with ultrafiltering techniques, and then purified by Sepharose 4 FF column. After detecting contents of antigen and protein, residual contents of bovine sera and cell DNA, and finishing other detections, the two sero - types of vaccines were mixed and the bivalent vaccine was elaborately prepared. Result According to the National Regulations for Biological Products and the essential standards of Killed Haemorrhagic Fever Vaccines for clinical trials, the purified bivalent Vero vaccine was stricdy and systematically detected.The quality was completely in accordance with the Regulations and the Standards . The vaccine is now ready for the approval of clinical trials. Conclusion The vaccine may be the substitution one for the exiting vaccines.

关 键 词：流行性出血热 双价纯化疫苗 VERO细胞 研究 

分 类 号：R512.8[医药卫生—内科学]