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目的评价产肠毒素大肠埃希菌(enterotoxigenic Escherichia coli,ETEC)K99-987P-F41融合蛋白在小鼠体内的免疫效果。方法制备ETEC融合蛋白K99-987P-F41亚单位疫苗、ETEC全菌体灭活疫苗、ETEC天然菌毛K99-987P-F41混合蛋白亚单位疫苗及ETEC重组菌毛K99-987P-F41混合蛋白亚单位疫苗,分别免疫小鼠,同时以免疫PBS的小鼠作为对照。ELISA法检测小鼠血清中IgG、IL-4和IFNγ的水平及小鼠粪便和小肠冲洗液中分泌型免疫球蛋白A(secreted immunogobulin A,sIgA)的含量;MTT法检测小鼠脾淋巴细胞增殖情况,并进行攻毒试验。结果 ETEC融合蛋白K99-987P-F41重组亚单位疫苗组小鼠血清中的抗ETEC的特异性IgG抗体、IL-4、IFNγ、粪便和小肠冲洗液中的sIgA、小鼠脾脏B和T淋巴细胞增殖能力均显著高于PBS对照组(P<0.05);ETEC融合蛋白K99-987PF41重组亚单位疫苗对小鼠的保护率为80%,与其他3种疫苗的小鼠保护率相近,而PBS对照组为0。结论 ETEC融合蛋白K99-987P-F41可有效诱导小鼠产生较高滴度的特异性抗体,可作为ETEC重组亚单位疫苗候选蛋白。
Objective To evaluate the immunogenicity of K99-987P-F41 fusion protein of enterotoxigenic Escherichia coli (ETEC) in mice. Methods ETEC fusion protein K99-987P-F41 subunit vaccine, ETEC whole cell inactivated vaccine, ETEC natural pili K99-987P-F41 mixed protein subunit vaccine and ETEC recombinant pili K99-987P-F41 mixed protein subunit Vaccine, respectively, were immunized mice, while immunized PBS mice as a control. The levels of IgG, IL-4 and IFNγ in sera of mice and the secreted immunogobulin A (sIgA) in mouse feces and intestinal fluid were detected by ELISA. The proliferation of mouse splenic lymphocytes was detected by MTT assay The situation, and conduct a challenge test. Results Anti-ETEC specific IgG antibodies, IL-4, IFNγ in sera from mice immunized with ETEC fusion protein K99-987P-F41 recombinant subunit vaccine, sIgA in feces and small bowel irrigation fluid, mouse spleen B and T lymphocytes (P <0.05). The protection rate of ETEC fusion protein K99-987PF41 recombinant subunit vaccine in mice was 80%, which was similar to that of the other three vaccines, while the PBS control Group is 0. Conclusion ETEC fusion protein K99-987P-F41 can effectively induce specific titers of antibodies in mice and can be used as a candidate protein of ETEC recombinant subunit vaccine.