Fusion protein of cytokine combined with chimeric antigen receptor and its application
A chimeric antigen receptor and fusion protein technology, applied in the field of biomedicine, can solve the problems of lack of safe and deliberate targets, complex tumor immune microenvironment, poor treatment effect, etc. Tumor effect, good anti-tumor effect
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[0070] 1 Construction of T2-mCAR molecules co-expressing IL-7 and CCL3
[0071] The mouse CAR gene constructed based on the retroviral vector backbone can be efficiently transduced into mouse T cells. Based on the original mouse second-generation CAR molecule T2-m28z, we constructed a co-expression mouse protein IL -7 and CCL3 mCAR fusion protein. Mouse IL-7 and CCL3 proteins are connected to the 3' end of the CAR gene without a stop codon by using a cleavable fusion protein adapter sequence (2A peptide, P2A). Since P2A can cut itself independently of proteases, The two structural domains are broken to achieve the result of co-expression of the three molecules. So far, we have constructed the murine CAR structure of T2-m28z IL7CCL3 ( 7P3 ), its amino acid sequence is shown in SEQ ID NO: 12, and its structure is shown in figure 1 shown.
[0072] 2 Preparation of CAR T cells co-expressing IL-7 and CCL3
[0073] The present invention prepares mCAR T cells according to the ex...
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