重组铜绿假单胞菌自组装纳米颗粒疫苗rePO-GroES的构建及免疫保护机制  

Recombinant self-assembled nanoparticle rePO-GroES confers protection against Pseudomonas aeruginosa in mice

作　　者：曾升 顾江 程新 魏玉洁 李宇航 高晨 崔红娟[1] 邹全明 ZENG Sheng;GU Jiang;CHEN Xin;WEI Yujie;LI Yuhang;GAO Chen;CUI Hongjuan;ZOU Quanming(Southwest University Medical Research Institute,Chongqing 400715,China;National Engineering Research Center of Immunological Products,College of Pharmacy,Army Medical University,Chongqing 400038,China)

机构地区：[1]西南大学医学研究院,重庆400715 [2]陆军军医大学药学与检验医学系国家免疫生物制品工程技术研究中心,重庆400038

出　　处：《免疫学杂志》2023年第8期696-701,713,共7页Immunological Journal

基　　金：国家重点研发专项(2021YFC2301405);国家自然科学基金(82271870)。

摘　　要：目的构建重组铜绿假单胞菌自组装纳米颗粒疫苗rePO-GroES,在肺炎模型中评价其保护作用并阐明其免疫保护机制。方法通过基因融合的方式,将编码铜绿假单胞菌保护性抗原PcrV-OprI(rePO)的编码基因连接至伴侣分子GroES基因的C端,构建并制备rePO-GroES蛋白;肌注rePO-GroES免疫小鼠后检测小鼠血清中特异性抗体、抗体亚型效价和脾细胞中分泌IFN-γ、IL-17A和IL-4的细胞比例;使用急性铜绿假单胞菌肺炎模型,评估rePO-GroES的保护效果。结果溶液中rePOGroES组装成分布均一的纳米颗粒,平均粒径约为28.2 nm;rePO-GroES免疫组小鼠生存率显著高于rePO免疫组和对照组,rePO-GroES免疫组体质量减轻幅度、感染状态评分、病理损伤程度、肺部细菌量和炎症反应均低于rePO免疫组和对照组;rePO-GroES免疫组小鼠血清中抗rePO IgGs滴度显著高于rePO免疫组和空白组,其中IgG1含量最高;rePO-GroES免疫组小鼠脾细胞中分泌IFN-γ、IL-17A和IL-4的细胞数均显著高于rePO免疫组和对照组。结论自组装纳米颗粒疫苗rePO-GroES安全性良好、免疫原性强、保护效果显著,是一个良好的铜绿假单胞菌候选疫苗。The aim of the study is to construct a recombinant Pseudomonas aeruginosa self-assembled nanoparticle vaccine rePO-GroES and to evaluate its protective effect in a pneumonia mouse model.The rePOGroES protein was constructed by fusing the gene encoding the protective antigen of Pseudomonas aeruginosa PcrVOprI(rePO)to the C-terminal of GroES.After mice were intramuscularly immunized with rePO-GroES,the titers of specific antibodies and antibody subtypes in sera and the proportion of cells secreting IFN-γ,IL-17A and IL-4 in spleen cells were detected.Furthermore,the protective effect of rePO-GroES was evaluated by using acute Pseudomonas aeruginosa pneumonia model.Data showed that rePO-GroES in aqueous phase was assembled into uniform nanoparticles with an average particle size of about 28.2 nm.The survival rate of mice vaccinated with rePO-GroES was significantly higher than that of mice immunized with rePO or PBS.The loss of body weight,infection score,pathological changes,lung bacterial load and inflammatory response in rePO-GroES-immunized mice were lower than those in the control group.The titer of rePO IgGs in sera of mice immunized with rePOGroES was significantly higher than that of mice immunized with rePO or PBS,and IgG1 was the predominant subtype.The level of splenocytes secreting IFN-γ,IL-17A and IL-4 in mice immunized with rePOGroES were significantly higher than those in the rePO group and control group.In conclusion,the self-assembled nanoparticle rePO-GroES has good safety and significant protective effect,which is a promising candidate vaccine for Pseudomonas aeruginosa.

关 键 词：铜绿假单胞菌 纳米颗粒 PcrV OprI GROES 

分 类 号：R392.9[医药卫生—免疫学]