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目的获得布鲁氏菌保护性抗原17.3kDa重组蛋白并以此构建亚单位疫苗。方法用PET32a(+)原核表达载体表达17.3kDa蛋白纯化后加氟氏佐剂肌肉注射免疫小鼠,三免后测定免疫功能进行免疫效果的评价。结果ELISA、WesternBlot检测到免疫鼠体内有特异性抗体产生,蛋白苗所诱导产生的IgG1抗体远远高于IgG2a;通过细胞因子和CD分子测定表明重组蛋白以诱发ThⅡ型免疫为主。结论所表达的布鲁氏菌17.3kDa蛋白苗具有诱导特异性细胞和体液免疫应答的能力,可作为潜在的布鲁氏菌新型疫苗或诊断抗原,有进一步研究的意义。
Objective To obtain a 17.3 kDa recombinant protein of Brucella protective antigen and construct a subunit vaccine. Methods Immunized mice were immunized with 17.3 kDa protein purified by PET32a (+) prokaryotic expression vector and intramuscularly injected with Freund ’s adjuvant. Immune function was evaluated after three immunizations. Results ELISA and Western blot showed that the specific antibody was produced in the immunized mice. The IgG1 antibody induced by the protein vaccine was much higher than that of the IgG2a antibody. The cytokines and CD molecules showed that the recombinant protein induced Th Ⅱ immunization. Conclusion The expressed 17.3 kDa protein vaccine of Brucella has the ability of inducing specific cellular and humoral immune responses and may serve as a potential new Brucella vaccine or diagnostic antigen for further research.