猴痘病毒蛋白的原核表达及免疫效果评价

Prokaryotic Expression and Evaluation of the Immune Effect of the Proteins of the Monkeypox Virus

猴痘是由猴痘病毒引起的一种人兽共患疾病。研发特异性预防和治疗猴痘的疫苗或药物成为目前全球研究重点之一。本研究以猴痘主要抗原蛋白其中的A29L、E8L、M1R为研究基础,加入His标签构建质粒、原核表达、纯化,并对表达蛋白进行SDS⁃PAGE及Western blot鉴定其特异性。后续将3种抗原蛋白与CpG⁃ODN 1826单佐剂、铝单佐剂或CpG与铝复合佐剂三次免疫小鼠,通过酶联免疫吸附实验及病毒蚀斑减少中和实验测定三价疫苗的结合抗体和中和抗体能力。结果表明,重组蛋白A29L、E8L、M1R诱导后均可溶性表达且表达量及纯度良好。三价疫苗以CpG⁃ODN 1826佐剂联合铝佐剂施行肌肉注射一组表现出较好的结合抗体和中和抗体能力。本研究为研发特异性针对猴痘的疫苗或药物提供基础。

Monkeypox is a zoonotic disease caused by infection with the monkeypox virus.The development of vaccines/drugs that prevent and treat monkeypox specifically has become a global research priority.In this study,A29L,E8L,and M1R,as the main antigen proteins of monkeypox,were used as the research subjects.A His tag was added to construct plasmids as well as to aid their prokaryotic expression and purification.The expressed protein was identified by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and western blotting.Subsequently,mice were immunized with these three antigen proteins combined with the adjuvant CpG⁃ODN 1826 or Al adjuvant.The ability to bind antibody and bind neutralizing antibody of the trivalent vaccine was determined by enzyme⁃linked immunosorbent assay and virus plaque reduction neutralization assay.The recombinant proteins A29L,E8L,and M1R were expressed in soluble form after induction,and the expression and purity were high.An intramuscular injection of a trivalent vaccine with CpG⁃ODN 1826 adjuvant combined with Al adjuvant showed a high ability to bind antibody and bind neutralizing antibody.This study provides a basis for the development of vaccines/drugs that target monkeypox specifically.