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以编码结核分枝杆菌分泌蛋白Ag85B、ESAT 6和MPT6 3的基因为插入片段 ,构建核酸疫苗联合免疫小鼠。以原核表达纯化的抗原为检测物 ,检测了抗原特异性的抗体和γ 干扰素的形成。研究表明 ,组合核酸疫苗第 3次免疫后 2 1天 ,实验小鼠血清中Ag85B抗体滴度达到 10 5以上 ,Ag85B抗原刺激产生的特异性γ 干扰素达到 (17.0± 7.0 )u/ml。组合疫苗虽然没有提高小鼠血清中ESAT 6及MPT6 3蛋白的特异性抗体滴度 ,但仍显著刺激产生了两种特异性的 γ 干扰素。攻毒实验表明 ,经组合疫苗免疫后小鼠肺部结核杆菌数量显著下降。肺部切片显示 ,免疫小鼠病理状况较对照组有明显改善。因此 ,研究提示Ag85B等组合核酸疫苗具有较好的结核病预防效果。
The DNA vaccine-co-immunized mice were constructed with the genes encoding Mycobacterium tuberculosis secreted proteins Ag85B, ESAT6 and MPT6 3 as inserts. The prokaryotic expression of the purified antigen as a test substance, detection of antigen-specific antibodies and interferon gamma formation. The results showed that the titer of Ag85B antibody in the serum of the experimental mice reached more than 105 on the 21st day after the third immunization with the combination nucleic acid vaccine and that of the Ag85B antigen reached (17.0 ± 7.0) u / ml. Although the combination vaccine did not increase the specific antibody titers of ESAT 6 and MPT6 3 proteins in mouse serum, two specific IFN-γs were still significantly stimulated. The challenge experiment showed that the number of M. tuberculosis in lungs of mice decreased significantly after the combination vaccine immunization. Lung sections showed that the pathological conditions in immunized mice were significantly improved compared with the control group. Therefore, the research suggests that the combination of Ag85B and other nucleic acid vaccine has a good TB prevention effect.