抗阿尔茨海默病重组腺相关病毒的制备与活性研究

Preparation and activity analysis of recombinant adeno-associated virus against Alzheimer’s disease

目的:制备可表达抗阿尔茨海默病单链抗体AT8-scFv的重组腺相关病毒AAV8-AT8-scFv,并探究其体内外活性。方法:利用293T细胞对重组病毒进行包装与纯化;SDS-PAGE、RT-qPCR对重组病毒的理化性质进行表征;细胞感染、Western blot法检测重组病毒的体外感染活性;以BALB/c小鼠为动物模型进行免疫,ELISA法、活体成像检测重组病毒的体内表达情况。结果:成功制备重组腺相关病毒AAV8-AT8-scFv,该病毒具有较好的感染能力,并可在小鼠体内长期稳定表达。结论:成功获得具有良好生物学活性的重组腺相关病毒,为阿尔茨海默病的免疫治疗提供新思路。

Objective:To prepare and analyze activity of a recombinant adeno-associated viral AAV8-AT8-scFv,which could express single-chain antibody AT8-scFv against Alzheimer’s disease.Methods:Recombinant viruses were obtained and purified using 293T cells;physical and chemical properties of the recombinant viruses were characterized by SDS-PAGE and RT-qPCR;In vitro infection activity of the recombinant virus was determined by cell infection assay and Western blot;BALB/c mice were used as animal model,and in vivo expression of the recombinant virus was detected by ELISA and vivisection imaging system.Results:AAV8-AT8-scFv was successfully obtained.The recombinant virus has good biological activity,and can be well expressed for a long time in mice.Conclusion:Recombinant adeno-associated virus with good biological activity was successfully obtained,which providing new ideas for immunotherapy of Alzheimer’s disease.