Structure optimization and function evaluation of chimeric antigen receptor T cells based on LCK recruitment and activation
10.13431/j.cnki.immunol.j.20240005
- VernacularTitle:基于LCK募集和活化的嵌合抗原受体T细胞的结构优化及其功能研究
- Author:
Lin ZOU
1
,
2
;
Siyin CHEN
;
Li DU
;
Kun TAO
Author Information
1. 400016,重庆医科大学基础医学院免疫学教研室
2. 400016,肿瘤免疫基础与转化研究重庆市重点实验室
- Keywords:
Chimeric antigen receptor T cell;
Lymphocyte-specific protein tyrosine kinase;
Cytotoxicity
- From:
Immunological Journal
2024;40(1):33-38
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to enhance the activation level and efficiency of second-generation chimeric antigen receptor-T(CAR-T)cells in killing tumor cells by modifying the lymphocyte-specific protein tyrosine kinase(LCK)structure of second-generation CAR,so as to lay the foundation for the preparation of novel second-generation CAR-T cells.Using genetic engineering techniques,lentiviral vectors for CD137-LCK2 CAR and CD137-PYAP2 CAR targeting human epidermal growth factor receptor 2(HER2)were constructed,and CD137-LCK2 and CD137-PYAP2 CAR-T cells were prepared by lentiviral packaging and infection of activated T cells.The differences between them in terms of CAR expression level,activating molecule CD 137 expression,phenotypic distribution,cytokines secretion,and killing effect on HER2-expressing tumor cells were detected and compared using flow cytometry,enzyme-linked immunosorbent assay,and cytotoxicity assay,respectively.Compared with the second-generation CD137 CAR-T cells,the structurally modified CD137-LCK2 and CD137-PYAP2 CAR-T cells targeting HER2 can increase the expression of activating molecule CD 137,increase the secretion of cytokine IFN-γ,and enhance the killing ability of target antigens;at the same time,these cells can be maintained in the memory state and can rapidly activate proliferation and differentiation after stimulation by antigen.In conclusion,CD137-LCK2 and CD137-PYAP2 CAR-T cells targeting HER2 are expected to activate and exert anti-tumor effects more effectively than second-generation CD 137 CAR-T cells.
