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目的用刚地弓形虫ROP2-P30复合重组蛋白疫苗免疫BALB/c小鼠观察对机体的影响,为研制安全有效的弓形虫疫苗奠定基础。方法PCR方法从刚地弓形虫基因组中扩增出ROP2和P30片段,将这两个片段分别亚克隆至pET-30a原核表达载体中,表达出ROP2-P30复合重组蛋白。用蛋白疫苗免疫BALB/c小鼠,ELISA检测免疫小鼠后体液中抗体和细胞因子的变化,观察攻击试验小鼠的的死亡率。结果ROP2-P30复合重组蛋白疫苗免疫BALB/c小鼠诱导机体产生大量的IgM、IgG抗体和IFN-γ、IL-2及IL-4细胞因子。攻击试验表明,复合重组蛋白免疫组和对照组相比,小鼠的存活时间明显延长。结论ROP2-P30复合重组蛋白免疫BALB/c小鼠诱导其产生高水平的体液和细胞免疫应答,该复合重组蛋白是抗刚地弓形虫感染的候选疫苗之一。
OBJECTIVE: To investigate the effects on organism of BALB / c mice immunized with ROP2-P30 recombinant recombinant protein vaccine and to lay a foundation for the development of a safe and effective Toxoplasma gondii vaccine. METHODS: ROP2 and P30 fragments were amplified from the genome of Toxoplasma gondii by PCR. The two fragments were subcloned into the prokaryotic expression vector pET-30a respectively to express the ROP2-P30 recombinant protein. BALB / c mice were immunized with protein vaccine. The changes of antibodies and cytokines in the humoral fluid after immunization were detected by ELISA and the mortality of mice in the challenge experiment was observed. Results The BALB / c mice immunized with ROP2-P30 recombinant recombinant protein vaccine induced a large amount of IgM, IgG antibodies and IFN-γ, IL-2 and IL-4 cytokines. The attack test showed that the survival time of the mice in the recombinant recombinant protein immunized group was significantly longer than that in the control group. Conclusion ROP2-P30 recombinant recombinant protein can induce high level of humoral and cellular immune responses in BALB / c mice. The recombinant recombinant protein is one of the candidate vaccines against T. gondii infection.