Protective efficacy of PLGA microspheres loaded with divalent DNA vaccine encoding the ompA gene of Aeromonas veronii and the hly gene of Aeromonas hydrophila in mice
Protective efficacy of PLGA microspheres loaded with divalent DNA vaccine encoding the ompA gene of Aeromonas veronii and the hly gene of Aeromonas hydrophila in mice
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DOI：
10.1016/j.vaccine.2013.08.053
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发表时间：
2013-11-19
期刊：
影响因子：
5.5
通讯作者：
Zhao, Baohua
中科院分区：
文献类型：
--
作者：
Gao, Shanshan;Zhao, Na;Zhao, Baohua
以乳酸-乙醇酸共聚物（PLGA）为载体，构建了编码维氏气单胞菌外膜蛋白A（ompA）和嗜水气单胞菌溶血素（hly）的二价融合DNA疫苗。将ompA基因和hly基因分别克隆到原核表达载体pET-28 a中，构建成重组质粒pET-28 a-ompA-hly。通过水包油包水（W/O/W）包封完成ompA-hly抗原模块在PLGA微球上的负载。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质印迹法检测pET-28 a-ompA-hly的分子量和特异性。该微球的平均粒径为100-150 μ m，负载效率（LE）为68.8%。通过腹膜内或胃内施用接受负载ompA-hly抗原的PLGA微球的小鼠产生强烈且持续的IgG应答，其显著高于（p < 0.05）单独pET-28 a-ompA-hly抗原所实现的应答。负载OmpA-hly抗原的PLGA微球疫苗独特地赋予针对攻击感染的持久（30天）无菌免疫。结果表明，ompA-hly抗原PLGA微球疫苗是一种合格的无菌保护性免疫载体系统。和A. Veronii感染。(C)2013爱思唯尔有限公司保留所有权利。
In the present study, poly (lactic-co-glycolic) acid (PLGA) was used as a carrier for a divalent fusion DNA vaccine encoding the Aeromonas veronii outer membrane protein A (ompA) and Aeromonas hydrophila hemolysins (hly) protein. The recombinant pET-28a-ompA-hly was constructed by inserting the ompA gene and hly gene into a pET-28a expression vector. Loading of ompA-hly antigen module on PLGA microspheres were accomplished by water-in-oil-in-water (W/O/W) encapsulation. The molecular weight and specificity of pET-28a-ompA-hly were detected by dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting. The microspheres showed an average particle size of 100-150 mu m and a loading efficiency (LE) of 68.8%. Mice received ompA-hly antigen-loaded PLGA microspheres by intraperitoneal or intragastric administration mounted strong and sustained IgG response, which was significantly higher (p < 0.05) than those achieved by pET-28a-ompA-hly antigen alone. OmpA-hly antigen-loaded PLGA microsphere vaccine uniquely conferred a long lasting (30 days) sterile immunity against challenge infection. Results indicated that ompA-hly antigen-loaded PLGA microsphere vaccine is a qualified candidate vector system for sterile protective immunity against A. hydrophila and A. veronii infections. (C) 2013 Elsevier Ltd. All rights reserved.