Construction and identification of a stable CT26 cell line expressing CD19-FLUC-GFP
Solid tumors lack well-defined targets for chimeric antigen receptor T-cell(CAR-T)therapy.Therefore,introducing a known target molecule,CD19,into solid tumor cell lines via lentiviral transduction to investigate the cytotoxicity of CD19 CAR-T cells can potentially support CAR-T cell therapy against solid tumors.In this study,a stable colon cancer CT26 cell line,CT26-CD19-FLUC-GFP,expressing CD19,firefly luciferase(FLUC),and green fluorescent protein(GFP),was constructed using a triple-plasmid lentiviral system.The growth characteristics of this cell line were consistent with those of the CT26 cell line.Subsequent flow cytometry analysis confirmed stable expression of CD19 and GFP in CT26-CD19-FLUC-GFP cells after serial passaging up to the 5th,10th,and 22nd generations.Further validation revealed significantly higher levels of CD19 mRNA and FLUC expression in CT26-CD19-FLUC-GFP cells continuously passaged up to the 22nd generation compared to the control CT26 cells.In comparison to T cells,CD19 CAR-T cells demonstrated substantial cytotoxicity against CT26-CD19-FLUC-GFP cells and MC38-CD19 cells.One week after intraperitoneal implantation of CT26-CD19-FLUC-GFP cells into mice,FLUC expression in the peritoneal region could be detected.These results indicate the successful establishment of a stable CT26 cell line expressing CD19-FLUC-GFP,which can be specifically targeted by CD19 CAR-T cells.
万方数据
维普
