The aim of this study was to investigate the functional characteristics and in vitro specific killing effect of EGFRvⅢ CAR-T cells co-expressing interleukin-15 and chemokine CCL19,in order to optimize the multiple functions of CAR-T cells and improve the therapeutic effect of CAR-T cells targeting EGFRvⅢ on glioblastoma(GBM).The recombinant lentivirus plasmid was obtained by genetic engineering,transfected into 293T cells to obtain lentivirus and infected T cells to obtain the fourth generation CAR-T cells targeting EGFRvⅢ(EGFRvⅢ-IL-15-CCL19 CAR-T).The expression rate of CAR molecules,proliferation,chemotactic ability,in vitro specific killing ability and anti-apoptotic ability of the fourth and second generation CAR-T cells(EGFRvⅢ CAR-T)were detected by flow cytometry,cell counter,chemotaxis chamber and apoptosis kit.The results showed that compared with EGFRvⅢ CAR-T cells,EGFRvⅢ-IL-15-CCL19 CAR-T cells successfully secreted IL-15 and CCL19,and had stronger proliferation,chemotactic ability and anti-apoptosis ability in vitro(all P<0.05),while there was no significant difference in killing ability in vitro.Therefore,CAR-T cells targeting EGFRvⅢ and secreting IL-15 and CCL19 are expected to improve the therapeutic effect of glioblastoma and provide an experimental basis for clinical trials.
chimeric antigen receptor T cellthe class Ⅲ variant of the epidermal growth factor receptorglioblastomainterleukin-15C-C chemokine ligand 19
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