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探讨载有日本脑炎病毒(Japanese encephalitis virus,JEV)prME蛋白与粒细胞-巨噬细胞集落刺激因子(Granulocyte-macrophage colony-stimulating factor,GM-CSF)编码基因重组子(命名为pJME/GM-CSF)的壳聚糖纳米颗粒的免疫佐剂效应及其机制。本研究采用复凝聚法制备壳聚糖-pJME/GM-CSF纳米颗粒;免疫组化法检测肌肉注射部位浸润细胞的类型,流式细胞仪检测不同免疫原免疫鼠后脾脏DC表型和功能的变化;乳酸脱氢酶释放法检测CTL活性。结果显示制备的壳聚糖-pJME/GM-CSF纳米颗粒鉴定正确;pJME/GM-CSF募集包括非成熟树突状细胞、巨噬细胞、粒细胞到注射部位,增加脾脏树突状细胞表面MHCII的表达,抗原摄取和递呈功能,增强疫苗诱导的细胞免疫反应,壳聚糖-pJME/GM-CSF纳米颗粒可进一步扩大pJME/GM-CSF的上述作用。这些结果表明壳聚糖可作为DNA疫苗肌注免疫时的传送载体,能够增强pJME/GM-CSF疫苗的细胞免疫应答。
To investigate the expression of prME protein of Japanese encephalitis virus (JEV) and granulocyte-macrophage colony-stimulating factor (GM-CSF) gene (named as pJME / GM- CSF) chitosan nanoparticle immunoadjuvant effect and its mechanism. In this study, chitosan-pJME / GM-CSF nanoparticles were prepared by complex coacervation, the type of infiltrating cells at the site of intramuscular injection was detected by immunohistochemistry, the phenotype and function of spleen DCs were detected by flow cytometry Changes; lactate dehydrogenase release assay CTL activity. The results showed that the prepared chitosan-pJME / GM-CSF nanoparticles were identified correctly; pJME / GM-CSF recruited include immature dendritic cells, macrophages and granulocytes to the site of injection and increased the surface MHCII of spleen dendritic cells The effect of pJME / GM-CSF on chitosan-pJME / GM-CSF nanoparticles can be further extended by the expression of the antigen, antigen uptake and presentation function, and the enhancement of the vaccine-induced cellular immune response. These results indicate that chitosan can be used as a delivery vehicle for intramuscular immunization of DNA vaccines and can enhance the cellular immune response of pJME / GM-CSF vaccines.