重组2型脊髓灰质炎病毒样颗粒的制备及免疫原性初步研究
Chinese Journal of Microbiology and Immunology(2021)
Abstract
目的:构建共表达P1和3CD的重组杆状病毒,利用昆虫细胞重组表达2型脊髓灰质炎病毒(poliovirus type 2,PV2)的病毒样颗粒(virus-like particle, VLP),制备PV2-VLP进行初步免疫原性研究。方法:基于粉纹夜蛾细胞(High 5)密码子偏好性,将PV2的P1基因和3CD基因进行序列优化,并对P1基因进行稳定性突变,构建突变型rBac-PV2-P1s-3CD和野生型rBac-PV2-P1-3CD杆状病毒。Western blot分别检测目的蛋白表达量,离子交换层析纯化PV2-VLP,透射电镜观察VLP高级结构确定组装效率。免疫大鼠评估免疫原性。结果:成功构建了可稳定表达P1s蛋白和3CD蛋白的重组杆状病毒。Western blot结果表明热稳定性突变后VLP产量相比野生型有所提高。电镜观察发现直径约30 nm的三维结构,表明VLP成功组装。动物实验结果表明,重组制备的PV2-VLP具有免疫原性,且能有效刺激产生中和抗体。结论:用昆虫细胞-杆状病毒表达系统可成功制备有效的VLP类疫苗,为PV-VLP疫苗的研制提供参考。
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Key words
Poliovirus,Virus-like particles,Insect cells,Recombinant baculovirus
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