Preparation method of CIK (cytokine-induced killer) blocked by PD-1 for cancer therapy

Abstract

The invention discloses a preparation method of CIK (cytokine-induced killer) blocked by PD-1 for cancer therapy and belongs to the field of medical treatment. The preparation method includes the coating step, the activating step, the cleaning step, the blocking step and the filtering step. In the coating step, an anti-human CD3 monoclonal antibody is coated in a cell culture flask; in the activating step, the CD3 monoclonal antibody, IFN gamma and IL-2 are utilized to activate and culture a lymphocyte; in the cleaning step, the lymphocyte which is activated and cultured by three times is cleaned; in the blocking step, the cleaned lymphocyte is suspended by normal saline to obtain suspended cell solution, the PD-1 monoclonal antibody is added into the suspended cell solution, and then suspending incubation is carried out; in the filtering step, the cell suspension obtained by incubation of the PD-1 monoclonal antibody is filtered and then is refilled with normal saline to obtain feedback cell suspension. The CIK blocked by PD-1 has the advantages of low consumption of the PD-1 monoclonal antibody, good blocking effect of PD-1 molecules, high killing activity and the like, and also has the advantages of operational simplicity, low preparation cost and convenience in clinical application.

Description

technical field [0001] The invention relates to the medical field, in particular to a method for preparing PD-1-blocked CIK for treating tumors. Background technique [0002] Programmed death receptor 1 (PD-1) is a protein molecule expressed on the surface of T cells. When PD-1 binds to its ligand PD-L1 or PD-L2, it can inhibit the transcription of downstream NF-κB genes, inhibit The secretion of interferon-δ, thereby inhibiting the killing activity of T cells. PD-L1 or PD-L2 is expressed in a variety of tumor cells, such as lung cancer, breast cancer, gastric cancer, esophageal cancer, hepatocellular carcinoma, malignant melanoma, ovarian cancer, pancreatic cancer, renal cell carcinoma and urothelial carcinoma and other tumor tissues High expression of PD-L1. The inhibition of T cell cytotoxic activity caused by the combination of PD-1 on the surface of T cells and PD-L1 expressed on tumor cells is one of the important mechanisms for tumor cells to escape immune surveilla...

AI Technical Summary

Problems solved by technology

During the culture of lymphocytes for treatment, the amount of culture medium is usually about 1500ml. The total volume of culture medium is relatively large, and the amount of PD-1 antibody is still relatively large. discard, cause a certain amount of waste

In addition, during the culture process, the secretions and metabolites of cells can affect the binding of PD-1 antibody to PD-1 to a certain extent, thus affecting the blocking effect of PD-1

[0004] The price of PD-1 antibody for treatment is expensive, adding PD-1 antibody during intravenous infusion or CIK culture, the disadvantages of large amount of PD-1 antibody, high cost, and easily affected PD-1 blocking effect have greatly affected Clinical application of PD-1 antibody

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