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为了解人类肠道病毒71型(EV71)重组保护性衣壳蛋白(rVP1)亚单位疫苗壳聚糖佐剂的作用,本研究利用rVP1蛋白与壳聚糖佐剂制备的疫苗口服免疫新西兰白兔,通过ELISA方法检测兔血清、粘膜洗液(小肠、鼻腔和肺)及粪便中特异性IgG、IgA抗体水平,中和试验检测血清中和抗体滴度,并通过抗原刺激体外培养的兔脾淋巴细胞分泌细胞因子(IFN-γ、IL-4)的水平评价rVP1口服免疫效果。结果显示,单纯口服rVP1蛋白仅可诱导产生较低水平的血清IgG和粘膜IgA抗体,而含佐剂疫苗组与单纯抗原免疫组差异显著,并可诱导产生中和抗体。细胞免疫检测结果显示rVP1含佐剂组兔脾淋巴细胞分泌IFN-γ和IL-4水平明显高于未加佐剂组。本研究为研制EV71VP1口服疫苗奠定了基础。
In order to understand the role of human recombinant enterovirus 71 (EV71) recombinant protective capsid protein (rVP1) subunit vaccine chitosan adjuvant, the present study orally immunized New Zealand white rabbits with a vaccine prepared by rVP1 protein and chitosan adjuvant The levels of IgG and IgA antibodies in rabbit serum, mucosal lotion (small intestine, nasal cavity and lung) and stool samples were detected by ELISA. Neutralization test was used to detect serum neutralizing antibody titers. Rabbit spleen lymphocytes The levels of cytokine secreted by cells (IFN-γ, IL-4) were evaluated for the effect of oral immunization with rVP1. The results showed that oral administration of rVP1 protein only induced low levels of serum IgG and mucosal IgA antibodies, while adjuvanted vaccine group and antigen-immunized group significant difference, and can induce the production of neutralizing antibodies. The result of cell immunization showed that the levels of IFN-γ and IL-4 secreted by spleen lymphocytes of rVP1-adjuvant group were significantly higher than that of the control group. This study lays the foundation for the development of EV71VP1 oral vaccine.