鼠伤寒沙门氏菌鞭毛蛋白可增强DnaJ蛋白的免疫反应  

Salmonella typhimurium Flagellin as adjuvants can enhance immune response of DnaJ protein

作　　者：严明[1] 刘宇思[1] 张帅[1] 胥文春[1] 王虹[1] 张雪梅[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国免疫学杂志》2014年第8期1023-1027,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.31070819);重庆市科委自然科学基金(No.cstc2012jjA10009)项目

摘　　要：目的:明确鞭毛蛋白的黏膜佐剂作用,并研究其与DnaJ蛋白联合免疫对肺炎链球菌感染的保护作用。方法:含有重组质粒pET-28(a)/鞭毛蛋白基因的大肠杆菌DE3经IPTG诱导后表达鞭毛蛋白,纯化后用于后期动物实验如下:将C57BL/6小鼠随机分为四组通过鼻腔分别滴入鞭毛蛋白(10μg)与DnaJ蛋白(10μg)混合剂(实验组)、DnaJ蛋白(10μg)(对照组1)、DnaJ蛋白(10μg)与GST蛋白(10μg)混合剂(对照组2),每组均30μl。通过检测抗体的效价、亚型以及细胞因子水平来评估鞭毛蛋白能否提高肺炎链球菌DnaJ蛋白诱导的免疫效应;然后用肺炎链球菌D39(5×107CFU)进行鼻腔攻毒,观察小鼠的生存率,以评估其保护效应。结果:实验组较对照组产生更高效价的血清IgG(其中主要产生IgG1)和更高水平的细胞因子IFN-γ、IL-17A和IL-4;另一方面实验组对致死性D39感染的保护率达60%,DnaJ蛋白免疫组的保护率为50%。结论:鞭毛蛋白与DnaJ蛋白联合应用可以增强宿主对DnaJ蛋白的免疫反应,并且对致死剂量的肺炎链球菌感染有保护作用,提示鞭毛蛋白可以作为肺炎链球菌DnaJ蛋白疫苗的佐剂应用。To make sure the role of flagellin as mucosal adjuvants and the protective effect of streptococcus pneumonia infection when combined with DnaJ protein.Methods：Recombinant plasmid pET-28 （ a）/flagella was transferred to E.coli BL21（DE3）.Over-expression of flagella was induced by IPTG and purification for animal study.All the C57BL/6 mice were randomly divided into three groups ,then respectively intranasally given the mixture of flagellin and DnaJ protein （ experimental group ） , DnaJ protein（ control group 1 ） and the mixture of GST and DnaJ protein （ control group 2 ）.The serum IgG and its subtype , cytokines secreted by mice spleen cells were detected by ELISA.At last all the C57BL/6 mice were intranasally immunized with Streptococcus pneumoniaD39.The protective effect by survival times were evaluated.Results： The mice of experimental group could secrete high level of serum IgG and cytokines IFN-γ,IL-4 and IL-17A.What more,the survival rate of mice in experimental group was 60%,a significant statistical difference with the control group.Conclusion：The flagellin as an adjuvant can reinforce the immune response of DnaJ protein and have better protection of resistance D 39 infection.We suggest that flagellin can be used as protein vaccine adjuvants.

关 键 词：鞭毛蛋白 DnaJ蛋白 肺炎链球菌 佐剂 

分 类 号：R378.14[医药卫生—病原生物学]