金黄色葡萄球菌GapC蛋白与鼠伤寒沙门氏菌Flic融合蛋白的构建与免疫原性研究  

Construction and immunogenicity of a fusion protein of Staphylococcus aureus GapC and Salmonella typhimurium Flic

作　　者：赵达[1] 王鹤[1] 梁宏儒[1] 胡旭[1] 姜东君[1] 尹辉[1] 高佳滨[1] 陈为宏[1] 乔波[1] 朱战波[1] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,大庆163319

出　　处：《中国人兽共患病学报》2014年第3期250-254,共5页Chinese Journal of Zoonoses

基　　金：黑龙江省重大科技攻关项目(No.GA09B301-2)~~

摘　　要：目的探讨金黄色葡萄球菌(Staphylococcus aureus,S.aureus)GapC蛋白与鼠伤寒沙门氏菌(Salmonella typhimurium,S.typhimurium)鞭毛蛋白融合蛋白的免疫保护作用。方法将纯化的Flic-GapC融合蛋白背部皮下接种免疫小白鼠,间隔3周免疫两次。应用间接ELISA方法测定小鼠血清中IgG抗体滴度;在二次免疫后2周,分离小鼠脾脏淋巴细胞,进行淋巴细胞增殖实验,利用Elispot方法检测IFN-γ和IL-4特异性分泌细胞;在二免后3周,用S.aureus Wood46株对免疫小鼠进行腹腔注射攻毒,观察1周并记录小鼠死亡数目。结果融合蛋白能够诱导小鼠产生特异性的免疫应答,二免后14d血清中IgG抗体滴度达到最高(1∶64 000),淋巴细胞刺激指数、分泌IFN-γ和IL-4的细胞数与对照组相比升高(P<0.05),对S.aureus的保护率达到70%。结论 Flic-GapC融合蛋白具有良好的免疫原性和免疫保护作用,可作为S.aureus的疫苗靶向进行深入研究。In order to investigate the immunoprotection of fusion protein of Staphylococcus aureus GapC protein and Salmonella typhimurium Flic protein, mice were immunized twice with purified Flic-GapC protein subcutaneously at a three week interval. The IgG antibody titers in immunized mice were measured with indirect ELISA. Two weeks after the secondary immunization, spleen lymphocytes were isolated to conduct lymphocyte proliferation experiment. Finally, the specific IFN-y- secreting ceils and IL-4-secreting cells were detected by Elispot assay. Three weeks after the secondary immunization, immu- nized mice were challenged with S. aureus strain Wood46 by intraperitoneal injection to evaluate the immunoprotection. After the one-week observation, the number of deaths in mice was recorded. Results showed that the Flic-GapC fusion protein could elicit mice producing specific immune response, and the antibody titer reached its peak at day 14 after boosted immunization （1 ： 64 000）. The spleen lymphocyte stimulation index as well as the cells secreting IFN-y and IL-4 increased significantly compared with that in the control group （P〈0.05）. Protection rate was 70% against S. aureus. The results above indicate that Flic-GapC fusion protein has a good immunogenicity and immunoprotection, and could become one S. aureus vaccine target in further study.

关 键 词：GapC蛋白 鞭毛蛋白 融合蛋白 免疫保护作用 

分 类 号：R378.1[医药卫生—病原生物学]