小分子药物提高慢病毒对人γδT细胞感染效率的研究  

The Study of Improving Efficiency of Lentivirus Infection in HumanγδT Cells with Small Molecular Drugs

作　　者：梁卉彤 林沁汝 杨辛毅 杨金龙 朱豫琪 梁玥 李敏[1] 成逸鹏 朱焕章[1] LIANG Huitong;LIN Qinru;YANG Xinyi;YANG Jinlong;ZHU Yuqi;LIANG Yue;LI Min;CHENG Yipeng;ZHU Huanzhang(School of Life Sciences,Fudan University,Shanghai 200438,China)

机构地区：[1]复旦大学生命科学学院,上海200438

出　　处：《复旦学报（自然科学版）》2024年第4期492-501,共10页Journal of Fudan University：Natural Science

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γδT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0.625μmol/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γδT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。γδT 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 inγδT cells,this study explored small molecule drugs that can improve lentivirus titer and lentivirus transduction efficiency inγδT 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 0.625μmol/L M344 treatment.TBK1/IKK epsilon inhibitor BX-795 increased the efficiency of lentivirus transduction inγδT cells,which raised from 3.5%to 10%when multiplicity of infection(MOI)was 5 without causing toxic effects onγδT cells.This study provides a new strategy for optimizing gene delivery based on lentivirus vector inγδT cells,and further promotes the application ofγδT cells in allogenic CAR cells therapy.

关 键 词：ΓΔT细胞 慢病毒载体 M344 诺考达唑 BX-795 

分 类 号：Q3[生物学—遗传学]