小分子药物提高慢病毒对人γδ T细胞感染效率的研究
The Study of Improving Efficiency of Lentivirus Infection in Human γδ T Cells with Small Molecular Drugs
梁卉彤 1林沁汝 1杨辛毅 1杨金龙 1朱豫琪 1梁玥 1李敏 1成逸鹏 1朱焕章1
作者信息
- 1. 复旦大学生命科学学院,上海 200438
- 折叠
摘要
γδT细胞识别靶细胞表面不依赖于主要组织相容性复合体(MHC)分子呈递的抗原,高效杀伤靶细胞且不引起移植物抗宿主病(GvHD),作为细胞平台在同种异体CAR细胞疗法开发中具有良好的应用前景.慢病毒(Lentivirus)载体是CAR-T细胞制备中常用的基因递送手段,为实现γδ T细胞中基于慢病毒载体的高效基因递送,本研究探索能够提高慢病毒滴度以及γδT细胞慢病毒感染效率的小分子药物.结果表明:组蛋白去乙酰化酶抑制剂M344和微管抑制剂Nocodazole可有效提高慢病毒滴度,其中当浓度为0.625 μmol/L时,M344可提高慢病毒滴度至2倍以上.TBK1/IKK epsilon抑制剂BX-795可提高γδ T细胞中慢病毒感染效率,当感染复数(MOI)为5时,BX-795可将感染效率从3.5％提升至10.7％,且未产生细胞毒性作用.本研究将为γδT细胞中基于慢病毒载体的基因递送优化提供一种新的策略,进一步推动γδT细胞在同种异体CAR细胞研究中的应用.
Abstract
γδ T cells can recognize antigens on target cells independently of major histocompatibility complex(MHC),kill target cells effectively without causing graft versus host disease(GvHD),which has great application prospect in allogenic CAR cell therapy development as platform.Lentivirus vector is a common gene delivery approach in CAR-T cells manufacture.To achieve efficient gene delivery based on lentivirus vector in γδ T cells,this study explored small molecule drugs that can improve lentivirus titer and lentivirus transduction efficiency in γδ T cells.The results showed that histone deacetylase inhibitor M344 and microtubule inhibitor Nocodazole could effectively improve lentivirus titer.Lentivirus titer was increased to more than 2 times with 0.625 μmol/L M344 treatment.TBK1/IKK epsilon inhibitor BX-795 increased the efficiency of lentivirus transduction in γδ T cells,which raised from 3.5％to 10％when multiplicity of infection(MOI)was 5 without causing toxic effects on γδ T cells.This study provides a new strategy for optimizing gene delivery based on lentivirus vector in γδ T cells,and further promotes the application of γδ T cells in allogenic CAR cells therapy.
关键词
γδT细胞/慢病毒载体/M344/诺考达唑/BX-795Key words
γδ T cells/lentivirus vector/M344/Nocodazole/BX-795引用本文复制引用
出版年
2024
NETL
NSTL
万方数据