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目的:完善A族链球菌(GAS)多价疫苗并探究其免疫保护效果。方法:构建GAS的多肽疫苗F7M6,将其中的M6多肽克隆作为对照,表达纯化后,将两种蛋白连同本室保存的GAS M1蛋白分别免疫小鼠,末次免疫后10 d,半数小鼠行ELISPOT检测IL-4和IFN-γ,ELISA检测各组的Ig G、Ig G1、Ig G2a;同时,以F7M6蛋白为诊断抗原检测其与抗链“O”阳性人血清的特异性结合;致死量的M1GAS攻毒并计算存活率。结果:无论是诱导细胞免疫,还是诱导抗体产生,F7M6组的水平均为最高;攻毒后F7M6蛋白的生存率为66.7%。F7M6蛋白检测抗链“O”阳性血清的阳性检出率为95%。结论:GAS的多肽疫苗F7M6能诱导特异性细胞和体液免疫应答,对GAS感染具有良好的保护效果,且F7M6的多价表位涵盖于时下流行的大部分GAS的血清型中。
Objective: To improve the polyvalent vaccine of Group A streptococci (GAS) and explore its immunoprotective effect. Methods: The GAS polypeptide vaccine F7M6 was constructed and the M6 polypeptide clone was used as a control. After expression and purification, the two proteins were immunized with the GAS M1 protein stored in our laboratory respectively. Ten days after the last immunization, half of the mice were subjected to ELISPOT IL-4 and IFN-γ were detected by enzyme-linked immunosorbent assay (ELISA). The Ig G, Ig G1, Ig G2a of each group were detected by ELISA. Meanwhile, the specific binding of anti-F7M6 protein to anti- The amount of M1GAS was challenged and the survival calculated. Results: The levels of F7M6 group were the highest in either induced cell or induced antibody. The survival rate of F7M6 protein was 66.7% after challenge. The positive detection rate of F7M6 protein detection anti-chain “O” positive serum was 95%. CONCLUSION: The GAS polypeptide vaccine F7M6 induces specific cellular and humoral immune responses with good protection against GAS infection, and the multivalent epitope of F7M6 is present in most serogroups of GAS that are prevalent today.