Specific cytotoxicity of a novel HER2-based chimeric antigen receptor modified T lymphocytes against HER2-positive tumor cells
10.3760/cma.j.issn.0529-5807.2017.10.011
- VernacularTitle: 靶向HER2嵌合抗原受体修饰T细胞的构建及其特异性杀伤肿瘤的作用
- Author:
Haijun TANG
1
;
Yuqin LIU
;
Xiaocui BIAN
;
Hailiang FENG
;
Pei GU
;
Hao SUN
;
Chunxia ZUO
;
Fangying ZHOU
;
Jing LIU
Author Information
1. Department of Pathology and Cell Resource Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China
- Publication Type:Journal Article
- Keywords:
Genes, erbB-2;
T-lymphocytes;
Molecular targeted therapy
- From:
Chinese Journal of Pathology
2017;46(10):714-720
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct the third generation chimeric antigen receptor based on a novel humanized anti-HER2 H1-2 scFv, and to investigate the specific cytotoxicity of H1-2 CAR modified T lymphocytes(CAR-T) against HER2+ tumor cells.
Method:The expression cassette of the third generation CAR gene and anti-HER2 H1-2 scFv were constructed and cloned into lentivirus transfer plasmid, and then the third generation H1-2 CAR was transduced into human T lymphocytes using lentivirus.Enzyme linked immunosorbent assay was used to detect the expression of cytokines IL2, and LDH release assay was used to detect the cytotoxic effect of the H1-2 CAR-T.Finally, NOD/SCID mice and HER2+ breast cancer cell line SKBR3 were used to detect the anti-tumor effect of H1-2 CAR-T in vivo.
Results:The third generation H1-2 CAR was successfully constructed.H1-2 CAR-T secreted high dose of IL2 after confrontation with HER2+ breast cancer cells.In vitro, the cytolytic rate of H1-2 CAR-T on high expression HER2+ tumor cells was significantly higher than that in low expression HER2 or non-expression HER2 tumor cells. At the efficacy to target ratio of 20, the cytolytic rate of H1-2 CAR-T against breast cancer cell SK-BR-3 could reach (90.1±2.8)%, while the cytolytic rate of H1-2 CAR-T against HER2- breast cancer cell MDA-MB-231 was only (13.5±4.7)%. In the mouse xenograft tumor model, H1-2 CAR-T cells inhibited breast cancer growth in vivo.At the end of the experiments, the average tumor weight in the H1-2 CAR-T cell treatment group was (0.7±0.1) g, the non-transfected T cell therapeutic group was (1.2±0.2) g, and the PBS group was (1.2±0.2) g. There was significant difference between the H1-2 CAR-T therapeutic group and the non-transfected T cell therapeutic group (P<0.05). However, there was no significant difference between the non-transfected T cell therapeutic group and the PBS treatment group (P>0.05).
Conclusion:The HER2-sepcific H1-2 CAR-T cells specifically kill HER2 positive cells, and further studies on CAR-T cells for the treatment of HER2+ cancers are useful.
