TCR<->(T Cell Receptor)/PD-1<-> double-negative T cell and construction method thereof

Abstract

The invention discloses a TCR<->(T Cell Receptor) / PD-1<-> double-negative T cell and a construction method thereof. Separated peripheral blood mononuclear cells are activated into T cells and TCRs and PD-1 are knocked out through a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) / Cas9 gene editing technology; the TCR<-> / PD-1<-> double-negative T cell is separated through a magnetic bead. The TCR<-> / PD-1<-> double-negative T cell prepared by the invention can be used for adoptive cell immunotherapies of tumors, including DC-CIK (Cytokine-Induced Killer), CTL (Cytotoxic T-Lymphocyte), TIL (Tumor Infiltrating Lymphocytes), CART (Chimeric Antigen Receptor Therapy) and the like. The TCR<-> / PD-1<-> double-negative T cell disclosed by the invention can be selected from other homologous and xenogenous healthy donators, so that the cell gets rid of limitation of current disease conditions of patients when being applied to the adoptive cell therapy; the cell can be prepared in a large batch in advance and the therapy cost is reduced.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a TCR- / PD-1- double negative T cell and a construction method thereof. Background technique [0002] With the development of immunology, tumor immunotherapy has made great progress. Tumor immunotherapy stimulates or mobilizes the patient's own immune system to enhance the anti-tumor immunity of the tumor microenvironment, thereby controlling and killing tumor cells. It is considered to be the fourth largest tumor treatment technology after surgery, radiotherapy and chemotherapy. In the past 20 years, tumor immunotherapy, represented by adoptive cell therapy (ACT) and immune checkpoint therapy (immune chenkpoint therapy), has made breakthrough progress, showing good application prospects, marking a new development in tumor therapy. The beginning of an era. [0003] Adoptive cellular immunotherapy (ACT) is to infuse activated and lethal immune cells to tumor patients, so that they can ...

AI Technical Summary

Problems solved by technology

In addition to the side effects such as cytokine storm caused by CART, there are still three major problems: first, for some advanced patients with low lymphocyte count or poor quality, the chance of CART treatment is lost; second, the efficacy of CART therapy in solid tumors Still not significant, probably due to the influence of immunosuppressive checkpoint signaling pathways, resulting in poor survival rate and low activity of immune cells in tumor tissue; finally, since CART is an individualized treatment, it is expensive and increases the burden on patients

However, there are still some problems in the existing adoptive cell immunotherapy or immunodetection blockade using PD1 antibody to treat tumors: (1) For some advanced patients with low lymphocyte count or poor quality, they lose the opportunity of CART treatment; ( 2) The efficacy of CART therapy in solid tumors is still not significant due to the influence of immunosuppressive checkpoint signaling pathways; (3) Both adoptive cell immunotherapy and immune checkpoint therapy are individualized treatments, which are expensive; (4) ) PD1 antibody causes autoimmune damage due to systemic administration

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