pVAX-Hsp70-HSV2gD DNA疫苗阳离子脂质体的制备  

Preparation of pVAX-Hsp70-HSV2gD DNA Vaccine Cationic Liposome

作　　者：樊建勇[1] 杨慧兰[1] 关蕾[1] 王颖[1] 仕瑶慧[1] 

机构地区：[1]广州军区总医院皮肤科,广东广州510010

出　　处：《中国皮肤性病学杂志》2006年第9期526-528,共3页The Chinese Journal of Dermatovenereology

基　　金：国家自然科学基金资助项目(编号:30371290);广东省医学科研基金资助课题(A2006492)

摘　　要：目的 制备pVAX-Hsp70-HSV2gD DNA疫苗阳离子脂质体，为基因免疫和基因治疗寻找一种简单、经济、有效的DNA投递系统。方法 采用逆向蒸发法，用卵磷脂、胆固醇、十八胺制备成阳离子脂质体包封pVAX-Hsp70-HSV2gD DNA疫苗，在透射电镜下观察脂质体颗粒的大小，应用紫外分光光度计检测DNA疫苗的包封率，免疫组化检测其转染真核细胞的效果。结果 制备的脂质体呈圆形、粒径大小较一致，平均粒径为580nm，DNA疫苗脂质体包封率达到50％～55％，能有效转染真核细胞。结论 成功制备了pVAX-Hsp70-HSV2gD DNA疫苗阳离子脂质体。该方法作为提高基因免疫效果的简单、经济、有效的手段之一，为其进一步的研究打下了基础。Objective To prepare pVAX-Hsp70-HSV2gD DNA vaccine cationic liposomes and obtain a better gene carrier to improve the effects of gene delivery. Methods Cationic liposomes made from lecithin, cholesterol and Eighteenth Amic by reverse evaporation technique were used for encapsulating pVAX-Hsp70-HSV2gD DNA vaccine. The shapes and sizes of the liposomes granule were observed by transmission electron microscope and the encapsulation efficiency of the liposome was detected by speetrophotometer. The expressed protein was characterized by immunochistocbemistry method. Results The liposomes were round and the diameter of granules was unanimous ,and the average size of complexes was 580 nm. The encapsulation efficiency of pVAX-Hsp70-HSV2gD DNA vaccine liposome reaches 50%～55%. And the DNA vaccine cationic liposome could be effectively transfeeted into mammalian cells. Conclusion The successful preparation and expression of pVAX-Hsp70-HSV2gD DNA vaccine cationic liposome laid a solid foundation for its further research and DNA cationic liposome complexes prepared by reverse evaporation technique could be applied in DNA delivery systerm.

关 键 词：单纯疱疹病毒 逆向蒸发 阳离子脂质体 包封率 DNA疫苗 

分 类 号：R394[医药卫生—医学遗传学]