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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 preparation of liposome-protamine-DNA complex (LPD) and the induction of dendritic cell (DC2.4) maturation. Methods Blank cationic liposomes were prepared by membrane-ultrasonic dispersion and incubated with protamine-DNA complex at room temperature to form LPD. The particle size and potential were determined. Flow cytometry was used to detect the expression level of mannose receptor in dendritic cell line DC2.4. The expressions of CD80, CD40, CD86 and MHC-2 on DC2.4 surface were detected to evaluate the effects of LPD on dendritic cells Mature induction. Results The optimal ratio of LPD was DDAB-protamine-DNA (2:1.5:1). The LPD particle size was 84.28 ± 0.56 nm and the zeta potential was 27.33 ± 1.23 mV. 83% of the mannose receptor expression on the DC2.4 surface was detected by the binding of FITC-ManBSA. LPD significantly upregulated the expression of DC2.4 surface marker (P <0.05). Conclusion LPD preparation process is simple, is a good vaccine adjuvant.