PEG3000修饰的脂质体作为流感疫苗佐剂的研究

Effect of PEG3000 modified liposomes as adjuvants for influenza vaccine

采用冻融-冻干法制备目标产品,用MTT法以刺激指数作为主要评价指标,通过单因素方差分析确定最佳处方;最佳处方制备的目标产品,通过腹腔免疫小鼠7、14和28 d后,用MTT法和表面标记法以刺激指数和CD4+/CD8+比值为评价指标进行细胞免疫原性研究;用微量血凝抑制法以抗体滴度比为评价指标检测体液免疫原性,探讨聚乙二醇(PEG)3000修饰的脂质体作为流感疫苗佐剂的最佳制备工艺和免疫原性。结果表明:PEG3000占卵磷脂的摩尔百分比为4%时,脂质体成膜后加入PEG3000制备的流感疫苗脂质体冻干粉为最佳处方;在1个免疫周期中, PEG3000修饰的流感疫苗脂质体组和其他组有显著差异(P<0.05)。这说明最佳处方制备的PEG3000修饰流感疫苗脂质体免疫原性增强且免疫时效延长。

The target product was prepared by freeze-thaw-lyophilization method,and the stimulation index(SI)was used as the main evaluation index by MTT method,and the best prescription was determined by one-way analysis of variance.The target product that was optimally formulated was intraperitoneally administered,and mice were immunized for 7,14,and 28 days,the cell immunogenicity was explored by the SI of MTT method and the CD4+/CD8+ratio of surface-labeled.Humoral immunogenicity using antibody titer ratio as an evaluation index was investigated by micro-hemagglutination inhibition method.This study explored the optimal preparation process and immunogenicity of PEG3000 modified liposomes as adjuvants for influenza vaccines.The results showed that the best preparation method was to add PEG3000 with a molar ratio of 4%to lecithin after liposome formation;the lyophilized powder of influenza vaccine liposome prepared by adding PEG3000 after liposome film formation was the best prescription.In an immunization period,the PEG3000 post-filming liposomes group was significantly different from the other groups(P<0.05).The data revealed that the PEG3000 modified influenza vaccine liposome immunity was significantly enhanced and prolonged immune circulation.