The invention relates to a preparation method of CAR (Cheimeric
Antigen Receptors)-T cells for co-expressing IL18 (
Interleukin 18) and CCL19 (
Chemokine C-C motif ligand 19)
protein and a targeting MUC1
gene. The preparation method comprises the following steps: transfecting DC<4+> / CD<+8> positive cells by
lentivirus containing IL18 and CCL19
protein coding sequences and targeting MUC1 CAR; screening and culturing amplified and transfected T cells to obtain the CAR-T cells for co-expressing IL18 and CCL19
protein and the targeting MUC1
gene. According to the preparation method provided by the invention, the proliferation capability and the life cycle of the CAR-T cells are enhanced through expressing third
cell signals IL18 and CCL19, improving the
osmosis effect of the CAR-T cells in solidtumors and maintaining the killing effect of the CAR-T cells on the
solid tumors. Furthermore, the co-expressed IL18 and CCL19 are connected through a 2A short
peptide and can be expressed respectively.