载铜绿假单胞菌DNA疫苗温敏水凝胶系统的构建及体内评价

石敏; 雍琴; 何颖娜; 黄仕琴; 赵轩; 余娴

doi:10.11665/j.issn.1000-5048.20210207

载铜绿假单胞菌DNA疫苗温敏水凝胶系统的构建及体内评价

1.
重庆医科大学附属第二医院Ⅰ期临床试验研究室，重庆 400010
2.
河北中医学院药学院，石家庄 050200
3.
重庆市江津区中心医院，重庆 402260

基金项目: 国家自然科学基金资助项目(No.82072327)；重庆市卫生计生委重点项目资助(No.2016ZDXM010)

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出版历程
- 收稿日期: 2020-11-03
- 修回日期: 2021-03-08
- 刊出日期: 2021-04-24

Construction and in vivo evaluation of a thermosensitive hydrogel system loading with Pseudomonas aeruginosa DNA vaccine

1.
Phase I Clinical Trial Center, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010
2.
College of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang 050200
3.
Department of Pharmacy, Central Hospital of Jiangjin District, Chongqing 402260, China

Funds: This study was supported by the National Natural Science Foundation of China (No. 82072327) and the Health Commission of Chongqing (No. 2016ZDXM010)

摘要

摘要: 构建载铜绿假单胞菌（Pseudomonas aeruginosa，PA） DNA疫苗的PLGA-PEG-PLGA温敏水凝胶系统，评价该系统的体内免疫效力。通过简单物理混合法制备PA DNA疫苗的PLGA-PEG-PLGA温敏水凝胶，测量其胶凝温度；评价其体外毒性、体外累计释放率；通过凝胶体积变化评价水凝胶的体内降解；将小鼠分为对照组（PBS）、水凝胶组（Hydrogel）、in vivo-jetPEI/质粒DNA组（in vivo-jetPEI/pDNA）、水凝胶+in vivo-jetPEI/质粒DNA组（Gel+in vivo-jetPEI/pDNA），免疫3次，每次间隔10 d，末次免疫2周后处死小鼠，检测血清特异性IgG抗体、脾淋巴细胞增殖情况及细胞上清液中IFN-γ水平。结果显示，PLGA-PEG-PLGA水凝胶具有温敏性，相变温度为（32 ± 0.5） ℃；体外对DC 2.4细胞无明显毒性；体外释放7 d时约释放80%的质粒DNA；PLGA-PEG-PLGA在体内具有可降解性，大约15 d几乎完全降解；体内免疫实验表明in vivo-jetPEI/pDNA及Gel+in vivo-jetPEI/pDNA组小鼠脾淋巴细胞明显增殖、特异性IgG抗体及IFN-γ水平升高，且水凝胶可增强DNA疫苗诱导的免疫反应水平。结果表明，载PA DNA疫苗的温敏水凝胶系统是研发PA疫苗有前景的一种新策略。
- PLGA-PEG-PLGA /
- 温敏水凝胶 /
- 铜绿假单胞菌 /
- DNA疫苗 /
- 免疫
Abstract: A thermosensitive hydrogel system consisting of PLGA-PEG-PLGA hydrogel and Pseudomonas aeruginosa DNA vaccine was constructed and the immune efficacy of the system was evaluated. The PLGA-PEG-PLGA thermosensitive hydrogel containing Pseudomonas aeruginosa DNA vaccine was prepared by a simple physical mixing method. The gelation temperature, cytotoxicity, and release curve in vitro were tested.The degradability of hydrogel in vivo was evaluated. The mice were divided into control group (PBS), hydrogel group (Hydrogel), in vivo-jetPEI/plasmid DNA group (in vivo-jetPEI/pDNA), and hydrogel + in vivo-jetPEI/plasmid group (Gel+in vivo-jetPEI/pDNA). Mice were immunized three times with a ten-day interval. Two weeks after the last immunization, the mice were sacrificed. The proliferation of splenic lymphocytes, serum specific IgG antibodies and IFN-γ in supernatant of splenic lymphocytes were detected. The gelation temperature of PLGA-PEG-PLGA hydrogel was (32 ± 0.5) ℃. There was no obvious toxicity to cells in vitro, and about 80% of plasmid DNA was released after 7 days in vitro. PLGA-PEG-PLGA hydrogel was biodegradable, and degraded almost completely after 15 days in vivo. The spleen lymphocytes proliferated; the levels of specific IgG antibodies and IFN-γ of in vivo-jetPEI/pDNA and Gel+in vivo-jetPEI/pDNA groups increased. The hydrogel could enhance the immune response induced by DNA vaccine.Results suggest that the thermosensitive hydrogel system consisting of PLGA-PEG-PLGA hydrogel and Pseudomonas aeruginosa DNA vaccine is a promising new strategy for the development of PA vaccine.
- PLGA-PEG-PLGA /
- thermosensitive hydrogel /
- Pseudomonas aeruginosa /
- DNA vaccine /
- immune

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参考文献(26)

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载铜绿假单胞菌DNA疫苗温敏水凝胶系统的构建及体内评价

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出版历程

Construction and in vivo evaluation of a thermosensitive hydrogel system loading with Pseudomonas aeruginosa DNA vaccine

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出版历程

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