论文部分内容阅读
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。
Objective To investigate the immunoprotection of GapC protein of Staphylococcus aureus (S. aureus) and S. typhimurium flagellin fusion protein. Methods Immunized mice were inoculated subcutaneously with purified Flic-GapC fusion protein twice a week for 3 weeks. The serum IgG antibody titers in mice were determined by indirect ELISA. Two weeks after the second immunization, the splenic lymphocytes were isolated and lymphocyte proliferation experiments were performed. The specific secretion of IFN-γ and IL-4 were measured by Elispot assay Immunized mice were intraperitoneally challenged with S. aureus Wood46 three weeks after the two immunizations and observed for one week and the number of mice killed was recorded. Results The fusion protein could induce specific immune response in mice. The highest IgG antibody titer in serum was 1:4 000 after 14 days of immunization (2). The number of lymphocyte stimulating index, the number of IFN-γ and IL-4 secreting The control group increased (P <0.05), the protection rate of S. aureus reached 70%. Conclusion Flic-GapC fusion protein has good immunogenicity and immunoprotection and can be used as a vaccine target for S. aureus for further study.