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目的为了应对流感流行多亚型并存的局面,进行了多表位核酸疫苗设计,并观察其免疫小鼠的细胞和体液免疫应答反应。方法设计并合成了含有H3、H9的HA和H5N1NA中和表位,M、NP基因保守序列CTL表位的多表位基因盒(Epi)。采用pVAX载体对H5HA、H7HA及Epi进行融合表达。经肌肉注射免疫6～8周龄雌性Balb/c小鼠,ELISA法检测小鼠血清中的针对H3579亚型流感抗体。三免后2周,分离脾淋巴细胞,进行T淋巴细胞转化试验和T淋巴细胞亚类数量检测。结果免疫小鼠获得了针对H3579亚型流感的体液和细胞免疫反应。结论完整HA抗原结合多表位的DNA疫苗模式的成功,预示了该DNA疫苗可能可以有效应对当前多种亚型流感并存的局面,并为其他种类流感疫苗的研究奠定了基础。
Objective To cope with the co-existence of influenza subtypes, the multi-epitope nucleic acid vaccine was designed and the cellular and humoral immune responses of the immunized mice were observed. Methods The multi-epitope gene box (Epi) containing H3, H9 HA and H5N1NA neutralizing epitopes and M, NP conservative sequence CTL epitopes was designed and synthesized. H5HA, H7HA and Epi were expressed by pVAX vector. Female Balb / c mice of 6-8 weeks old were immunized by intramuscular injection, and antibodies against H3579 subtype influenza in mice serum were detected by ELISA. Two weeks after the three immunizations, splenic lymphocytes were isolated and tested for T lymphocyte transformation and T lymphocyte subsets. Results The mice were immunized with the humoral and cellular immune responses against H3579 subtype influenza. Conclusion The success of the DNA vaccine model with intact HA antigen and polyepitope indicates that the DNA vaccine may effectively cope with the current coexistence of multiple subtypes of influenza and lays the foundation for the research of other kinds of influenza vaccines.