Vero细胞制备流感病毒疫苗

Preparation of Influenza Vaccine with Vero Cells

目的利用转瓶培养Vero细胞制备流感病毒疫苗。方法将流感病毒接种于Vero细胞上培养,优化培养条件,收获的病毒液经灭活、超滤浓缩和层析纯化,检测病毒的血凝素(HA)滴度及血凝素含量。按此工艺制备流感疫苗,根据血凝素含量稀释为不同浓度,免疫小鼠,观察不良反应,并通过血凝抑制(HI)试验检测抗体水平。结果Vero细胞培养流感病毒的最佳条件为:维持液中胰酶浓度12.5～15μg/ml,pH值7.4～7.6,Vero细胞经多次洗换后再接种病毒,病毒在细胞上培养72～96h收毒。所有免疫小鼠均未出现不良反应,小鼠抗体阳转率均为100%,HI抗体滴度均不低于同剂量的阳性对照组。结论Vero细胞可用于流感病毒的培养和制备疫苗。

Objective To prepare influenza vaccine with Vero cells cultured in rolling bottle.Methods Influenza virus was inoculated onto Vero cells cultured in rolling bottles,and the condition for culture was optimized.The harvested virus liquid was inactivated,concentrated by ultrafiltration,purified by chromatography and determined for HA titer and content.Influenza vaccine was prepared by the procedure and diluted to various concentrations according to HA content,then used for immunization of mice.The immunized mice were observed for adverse reactions and determined for antibody levels by HI test.Results The condition for culture of influenza virus in Vero cells was optimized as follows：the trypsin concentration in maintain medium was 12.5～15 μg /ml;the pH value of maintain medium was 7.4 ～ 7.6;the virus was inoculated onto Vero cells after changing the maintain medium for several times and cultured for 72 ～ 96 h then harvested.No adverse reactions were observed in the immunized mice.All the antibody positive conversion rates of the mice were 100%.The HI titers of antibody in the mice immunized with the prepared vaccine were significantly higher than those with commercial vaccine.Conclusion Vero cells may be used for culture of influenza virus and preparation of influenza vaccine.