Method for rapidly preparing PD-1<->T cells by high-efficiency gene editing and application thereof

Abstract

The invention discloses a method for obtaining PD-1<->T cells. The method comprises the following steps: (1) obtaining human peripheral blood T-lymphocytes; (2) performing plasmid transfection; (3) preparing fusion cells; (4) amplifying PD-1<->T tumour specific T cells. The invention also discloses application of the method for obtaining the PD-1<->T cells. The PD-1<->T cells obtained by utilizingthe method disclosed by the invention excludes a negative inhibitory regulatory receptor, has a very good anti-tumour effect, effectively prolongs the survival time of a tumour-bearing mouse comparedwith traditional adoptive therapy cells, and is suitable to be popularized in medical field.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for rapidly preparing PD-1-T cells by efficient gene editing and its application. Background technique [0002] T cells in the body's immune system are the main component of lymphocytes. T lymphocytes are mainly used to resist foreign invaders and have various biological functions. It is the body's main "warrior" against disease invasion, infection and tumor formation. At present, the methods of tumor treatment are generally surgery, radiotherapy and chemotherapy. There are some problems in these methods, among which immune escape phenomenon will appear in adoptive therapy, which will affect the anti-tumor effect. [0003] PD-1 (programmed death 1), the programmed death receptor 1, is a negative inhibitory regulatory receptor. When T cells are activated, the expression of PD-1 can be significantly increased. Through the combination with the ligand PD-L1 The effec...

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Problems solved by technology

[0005] However, although the existing CRISPR / Cas9 gene technology has been applied in many fields, it has also shown good results, but in terms of tumor treatment, especially knocking out the PD-1 gene on T cells is not in the public technology. Not involved, in addition, the existing CRISPR / Cas9 knockout PD-1 gene technology did not find the combined application of high penetration peptide small molecule TAT, so there are problems of low plasmid transfection efficiency and PD-1 gene editing efficiency

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