摘要
目的研制新的流行性乙型脑炎传代细胞疫苗。方法将流行性乙型脑炎弱毒株ＳＡ１４１４２适应于Ｖｅｒｏ细胞，作纯化灭活疫苗，比较不同培养方法病毒在Ｖｅｒｏ细胞的增殖规律，对病毒的浓缩和纯化条件进行研究，并将所制备的疫苗按不同蛋白浓度免疫动物。结果发现普通转瓶培养，可长时间维持病毒的高滴度。在感染病毒后，以无牛血清的培养基替代含牛血清的培养基，不仅可维持病毒的高滴度增殖，而且可多次收获，确立了应用８％ＰＥＧ８０００浓缩病毒液，１５％～６０％的蔗糖密度梯度超速离心纯化的工艺，每剂量为０５μｇ纯化疫苗的中和抗体水平，即可达到与现有商品疫苗相一致的滴度。结论ＳＡ１４１４２株制备的Ｖｅｒｏ细胞纯化疫苗，有望成为一种新的反应轻、效价高的疫苗。
Objective To develop a kind of new Japanese Encephalitis(JE) vaccine prepared on Vero cells. Methods A JE attenuated virus strain SA14142 was adapted on Vero cells for preparation of purified inactivated vaccine. Comparison of the growth curves of SA14142 in roller bottle and in spinner flask was made.After the virus inoculation and absorption on Vero cells for 2 hours, cultures were replaced with serumfree MEM, the culture supernatants were harvested on day 2,4,6 after inoculation. The virus fluids were pooled, concentrated by 8% PEG, and purified on 15%60% sucrose density gradients. The purified virus was inactivated with 0.02% formalin. Results It showed that virus titer was higher and maintained longer in roller bottle. Mice vaccinated twicely with 0.5 g dose of the purified inactivated vaccine induced neutralizing antibody titers equal to that of the mice vaccinated with primary hamster kidney inactivated vaccine. Conclusion This Vero cell prepared purified and inactivated JE vaccine made by SA14142 strain could be used for human as a kind of new JE vaccine.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
1999年第2期191-193,共3页
Chinese Journal of Experimental and Clinical Virology
关键词
流行性乙型脑炎
VERO细胞
疫苗
弱毒株
Japanese encephalitis(JE)Vero cellsVaccineJE attenuated virus strain
