站内搜索
最新录用更多>>
DC靶向适配体修饰的载铜绿假单胞菌DNA疫苗递送系统的构建及体外评价
    点此下载全文
引用本文:黄仕琴,石敏,何颖娜,岳瀚勋,余娴.DC靶向适配体修饰的载铜绿假单胞菌DNA疫苗递送系统的构建及体外评价[J].中国药科大学学报,2019,50(6):743-752.
Cite:HUANG Shiqin,SHI Min,HE Yingna,YUE Hanxun,YU Xian.Construction and in vitro evaluation of DC-targeted aptamer-modified Pseudomonas aeruginosa DNA vaccine delivery system[J].J China Pharm Univ,2019,50(6):743-752.
摘要点击次数: 56
全文下载次数: 58
作者单位
黄仕琴 重庆医科大学附属第二医院I期临床试验研究室 
石敏 重庆医科大学附属第二医院I期临床试验研究室 
何颖娜 河北中医学院药学院 
岳瀚勋 重庆医科大学药学院生物化学与分子药理学重点实验室 
余娴 重庆医科大学附属第二医院I期临床试验研究室 
基金项目:重庆市卫生计生委重点项目资助(No.2016ZDXM010)
中文摘要:构建了一种DC靶向适配体修饰的铜绿假单胞菌(Pseudomonas aeruginosa,PA)DNA疫苗递送系统。采用乙醇注入法制备阳离子脂质体,静电吸附法制备载pVAX1-OprF-VP22的阳离子脂质体(Lip-pOprF-VP22),探讨不同DOTAP/pDNA质量比的Lip-pOprF-VP22对pVAX1-OprF-VP22的包封效果、对DC2.4的细胞毒性及转染率,筛选最佳质量比的Lip-pOprF-VP22测定其粒径及Zeta电位;后插法制备DC靶向适配体修饰的载pVAX1-OprF-VP22的阳离子脂质体(Apt-Lip-pOprF-VP22),检测其转染DC2.4后OprF蛋白的表达量及对小鼠骨髓来源树突状细胞(bone marrow-derived dendritic cells,BMDCs)成熟的影响。结果表明,Lip-pOprF-VP22随着DOTAP/pDNA质量比增加包封率逐渐增加,当质量比为5∶1时即能很好的包封pVAX1-OprF-VP22;当Lip-pOprF-VP22作用于DC2.4 24 h或48 h后,不同质量比的Lip-pOprF-VP22对DC2.4的存活率均在80%以上;当DOTAP/pDNA质量比由2∶1增加到10∶1,转染率表现为先增加、后降低的趋势,其中DOTAP/pDNA质量比为4∶1、5∶1时转染率相对较高;当DOTAP/pDNA质量比为5∶1时,Lip-pOprF-VP22粒径为(171.67±1.27)nm,Zeta电位为(11.30±0.57)mV;Apt-Lip-pOprF-VP22转染DC2.4后可表达更多OprF蛋白且可明显促进BMDCs的成熟。
中文关键词:铜绿假单胞菌  DNA疫苗  DC靶向  适配体  OprF
 
Construction and in vitro evaluation of DC-targeted aptamer-modified Pseudomonas aeruginosa DNA vaccine delivery system
Abstract:This study aimed to construct a DC-targeted aptamer-modified Pseudomonas aeruginosa(PA)DNA vaccine delivery system. The cationic liposome was prepared by ethanol injection method. The cationic liposome loading pVAX1-OprF-VP22(Lip-pOprF-VP22)was prepared by electrostatic adsorption method. The encapsulation efficiency of Lip-pOprF-VP22 with different mass ratios of DOTAP/pDNA on pVAX1-OprF-VP22, cytotoxicity and transfection rate to DC2. 4 in vitro were discussed. The particle size and zeta potential of Lip-pOprF-VP22 with best mass ratio were tested. Aptamer-modified cationic liposome loading pVAX1-OprF-VP22(Apt-Lip-pOprF-VP22)was prepared by post-insertion method. The expression of OprF protein after transfection of DC2. 4 and its effect on the maturation of bone marrow-derived dendritic cells(BMDCs)were detected. Data showed that as the mass ratio of DOTAP/pDNA increased, the encapsulation efficiency of Lip-pOprF-VP22 on pVAX1-OprF-VP22 was gradually increased. When the mass ratio was 5 ∶1, pVAX1-OprF-VP22 was encapsulated well. When Lip-pOprF-VP22 with different mass ratios was applied to DC2. 4 for 24 h or 48 h, the survival rates of DC2. 4 were all above 80%. When the mass ratio of DOTAP/pDNA increased from 2 ∶1 to 10 ∶1, the transfection rate increased first and then decreased. When the mass ratios of DOTAP/pDNA were 4 ∶1 and 5 ∶1, the transfection rates were relatively high. When the mass ratio of DOTAP/pDNA was 5 ∶1, the particle size of Lip-pOprF-VP22 was(171. 67±1. 27)nm, and the Zeta potential was(11. 30±0. 57)mV. Furthermore, Apt-Lip-pOprF-VP22 can express more OprF protein and significantly promote the maturation of BMDCs. In conclusion, Apt-Lip-pOprF-VP22 can target to DC and promote the maturation of DC.
keywords:Pseudomonas aeruginosa  DNA vaccine  DC-targeted  aptamer  OprF
查看全文  查看/发表评论  下载PDF阅读器
分享按钮