靶向DC DNA疫苗的研制及抗鼠眼角膜HSV-1感染的初步研究  

Experimental study of construction of DC-targeting DNA vaccine and its prophylactic effect on herpes simplex keratitis

作　　者：汤如[1] 胡凯[2] 

机构地区：[1]东南大学医学院眼科学系,江苏南京210009 [2]南京大学医学院附属鼓楼医院眼科,江苏南京210008

出　　处：《现代医学》2016年第11期1546-1550,共5页Modern Medical Journal

摘　　要：目的:构建靶向树突状细胞(DC)的DNA疫苗pRSC-NLDC145·gD-IL-21,针对抵抗鼠眼角膜单纯疱疹病毒1型(HSV-1)的感染,探讨其是否能产生更强的免疫应答反应。方法:通过基因克隆的方法构建靶向DC的重组质粒pRSC-NLDC145·gD-IL-21,对其进行鉴定,并和壳聚糖包裹组成纳米DNA疫苗,分别以靶向组pRSC-NLDC145·gD-IL-21+壳聚糖、非靶向组pRSC-gD-IL-21+壳聚糖、pRSC+壳聚糖和壳聚糖免疫小鼠3次,间隔2周,检测各疫苗对小鼠免疫应答的水平。结果:靶向DC的重组质粒可在真核细胞内进行蛋白水平的表达,且和纳米壳聚糖包裹形成包封率较高的靶向DNA疫苗组,较非靶向组在鼠体内产生了更强的免疫应答反应。结论:眼表黏膜接种靶向DC的纳米DNA疫苗pRSC-NLDC145·gD-IL-21,在小鼠单纯疱疹病毒性角膜炎(HSK)动物模型中显示了更强的免疫保护反应。Objective： To construct DC-targeting DNA vaccine pRSC-NLDC145·gD-IL-21 and to study its ability to induce immune response and to resist the HSV-1 infection of cornea in mice. Methods： The recombinant plasmid of DC-targeting DNA vaccine pRSC-NLDC145·gD-IL-21 was constructed through gene cloning method. After being identified by molecular methods, we constructed chitosan nanoparticles, and then the vaccine pRSC- NLDC145 · gD- IL- 21 ＋ chitosan, pRSC- gD- IL- 21 ＋ chitosan, pRSC ＋ chitosan and chitosan were respectively inoculated into BALB/c mice for 3 times with 2 weeks interval. The levels of immune response were detected 2 weeks after the last immunization. Results： The data presented showed that the targeted genes were successfully expressed in the transfected 293T cells. The agarose gel electrophoresis analysis confirmed that the nanoparticles could be combined with plasmids through electrostatic interactions. Compared with the control mice, the DC- targeting DNA vaccine pRSC-NLDC145·gD-IL-21 ＋ Chitosan induced mice generated stronger immune response. Conclusion： These findings indicate that the DC- targeting DNA vaccine pRSC- NLDC145·gD- IL- 21 ＋ chitosan may induce a stronger immune response in immunized mice.

关 键 词：单纯疱疹病毒1型 树突状细胞靶向DNA疫苗 壳聚糖 单纯疱疹病毒性角膜炎 小鼠 

分 类 号：R772.21[医药卫生—眼科] R392-33[医药卫生—临床医学]