脂质体-鱼精蛋白-DNA复合物的制备及对树突状细胞的成熟诱导  

Preparation of liposomes-protamine-DNA complexes for maturation induction on dendritic cells

作　　者：李攀[1] 陈思穆[1] 龚涛[1] 张志荣[1] 孙逊[1] 

机构地区：[1]四川大学华西药学院靶向药物与释药系统教育部重点实验室,四川成都610041

出　　处：《华西药学杂志》2013年第3期246-248,共3页West China Journal of Pharmaceutical Sciences

摘　　要：目的研究脂质体-鱼精蛋白-DNA复合物(LPD)的制备方法及对树突状细胞(DC2.4)的成熟诱导作用。方法薄膜-超声分散法制备空白的阳离子脂质体,再与鱼精蛋白-DNA复合物室温孵育形成LPD;测定其粒径和电位。流式细胞仪检测鼠源树突状细胞系DC2.4的甘露糖受体表达水平;用DC2.4表面标记分子CD80、CD40、CD86和MHC-2的表达水平,考查LPD对树突状细胞的成熟诱导作用。结果 LPD的最佳比例为DDAB-鱼精蛋白-DNA(2∶1.5∶1);LPD粒径为84.28±0.56 nm,Zeta电位为27.33±1.23 mV。通过FITC-ManBSA的结合作用检测到DC2.4表面有83%的甘露糖受体表达。LPD明显上调DC2.4表面标记分子的表达水平(P<0.05)。结论 LPD制备工艺简单,是一个良好的疫苗佐剂。OBJECTIVE To study the maturation induction of preparation of liposomes-protamine-DNA complexes（LPD） on dendritic cells（DC2.4）.METHODS Thin film-sonication dispersion technology was used to prepare blank cationic liposomes,which were then incubated with protamine-DNA complexes at room temperature to form LPD.Size distribution and Zeta potential were measured respectively.The expression of mannose receptor on dendritic cells was confirmed by ligand assay using flow cytometry.The maturation activation ability of LPD was performed on dendritic cells（DC2.4） by detecting the expression level of cell surface markers,including CD80,CD40,CD86 and MHC-2 using flow cytometry.RESULTS The optimized LPD composed of DDAB-protamine-DNA with a ratio of 2：1.5：1 had a size of 84.28±0.56 nm and Zeta potential of 27.33±1.23 mV.High expression（83%） of mannose receptor on DC2.4 cell surface was confirmed by binding assay of FITC-ManBSA.More importantly,LPD significantly enhanced the expression of cell surface makers of DC2.4.CONCLUSION LPD prepared by a convenient method is a potent immunoadjuvant.

关 键 词：脂质体-鱼精蛋白-DNA复合物 树突状细胞 成熟诱导 

分 类 号：R94[医药卫生—药剂学]