The specific killing effect of anti-CD19 CAR-T cells on K562CD19+tumor cells
Objective To construct a lentivirus vector expressing anti-CD19 chimeric antigen receptors and its engineered T cells and investigate the specific cytotoxicity of CAR-T cells on CD19 positive B-cell lymphoma cells. Methods Technology of molecular cloning was employed to construct anti-CD19 CAR with self-cleavage enhanced green fluorescent protein, and then to package lentivirus with packaging and envelope plasmids. Using lentivirus delivery system, anti-CD19 CAR were overexpressed on primary-T cells. Followed by the identify of anti-CD19 CAR expression on T cells using flow cytometry, ELISA and LDH assay were used to detect the IL-2 secretion and killing effect of the CAR-T cells, respectively. Results We successfully constructed the CD19-specific CAR gene recombined lentivirus and established CD19-specific CAR-engineered T cells. Results from flow cytometry evaluated with EGFP suggested that anti-CD19 CAR were stably and efficiently expressed on T cell surface. The secretion of IL-2 from CAR-T cells was significantly more than the primary-T cells(P < 0.01), and the cell killing activity of CAR-T cells increased with the E:T change, especially the killing effect rate over 50% at E:T = 10:1 (P < 0.05). Conclusion We successfully establish the CD19-specific CAR-engineered T cells with specifically and efficiently killing effect on the target cells.
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