Structure optimization and function evaluation of chimeric antigen receptor T cells based on LCK recruitment and activation
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.
Chimeric antigen receptor T cellLymphocyte-specific protein tyrosine kinaseCytotoxicity
万方数据
维普
