中国肿瘤生物治疗杂志2018，Vol.25Issue(4)：389-393,5.DOI:10.3872/j.issn.1007-385x.2018.04.012
嵌合抗原受体修饰的CD19-CAR-T的体外构建、扩增及初步功能鉴定
In vitro construction and amplification and primary functional analysis of antiCD19 chimeric antigen receptor (CD19-CAR) modified T cells
摘要
Abstract
Objective:To establish a chimeric antigen receptor (CAR) modified T cells specifically targeting CD19 molecule (CD19-CAR-T cells) and to testify their in vitro killing effect on target cells.Methods:CD19-CAR fragments yielded by PCR were constructed into pCDH-GFP lentiviral vectors by molecular cloning technology.The packaged lentiviral particles were transducted into CD3+ T cells of donors.Transduction efficiency was measured by flow cytometry and PCR.The in vitro cytotoxicity of obtained CD19-CAR-T cells against CD19+ Ramos cells was tested by 7-AAD staining.Results:The amplification folds of CD3+ T cells increased to (78.8± 23.2) folds after in vitro culture for 10 days,and about (58.3±5.4)％ cells expressing GFR About (57.4±9.3)％ CD19+Ramos cells were specifically killed by the CD 19-CAR-T cells in vitro at the E∶T ratio of 5∶ 1.Conclusion:This study successfully established an effective method for constructing and amplifying CD19-CAR-T cells in vitro,which showed profound efficiency and specific cytotoxity against CD19+ Ramos cells.And this report might provide an experimental evidence for clinical treatment ofCD19+ B cell neoplasmas.关键词
CD19嵌合抗原受体/白血病/淋巴瘤/细胞杀伤Key words
CD19 chimeric antigen receptor/leukemia/lymphoma/cytotoxicity分类
生物科学引用本文复制引用
李剑,刘根桃,朱学军,田芳,姜鹏君,孔祥图,吴坚,殷婷婷,邢芸,金亮,郝瑞栋..嵌合抗原受体修饰的CD19-CAR-T的体外构建、扩增及初步功能鉴定[J].中国肿瘤生物治疗杂志,2018,25(4):389-393,5.基金项目
江苏省社会发展-临床前沿技术资助项目(No.BE2016809) （No.BE2016809）
南京市科技发展计划项目资助(No.201503011).Project supported by the Foundation of the Jiangsu Social Development-Clinical Frontier Technology (No.BE2016809),and the Foundation of the Nanjing Science and Technology Development Plan (No.201503011) （No.201503011）
