Preparation method and application of autologous CAR (chimeric antigen receptor)-T cell

Abstract

The invention relates to a preparation method and application of an autologous CAR (chimeric antigen receptor)-T cell. An established CD28-CD137-CD19-CD3 full-length gene is guided into a T-cell of a patient by a CRISPR / Cas9 technology to prepare the CAR-T cell, and the CAR-T cell is subjected to expansion in vitro and then returns in the body of the patient to perform anti-tumor treatment. Compared with the traditional tumor treatment method, the method has the advantages that the method is cell targeted therapy and small in side effect; the gene modified T cell can stably express an antigen binding domain on the surface and identify a target antigen, and does not have MHC limit; and the tumor treatment effect is improved.

Description

technical field [0001] The invention relates to the fields of medicine, immunology, cell biology and molecular biology, in particular to a method for preparing autologous CAR-T cells and its application. Background technique [0002] As a new cancer treatment method, immune cell therapy has been clinically proven more and more. CAR-T cell (Chimeric Antigen Receptor T cell) technology uses the patient's own immune cells to eliminate cancer cells. Cellular immune cell therapy has shown extremely high advantages in the treatment of a variety of blood cancers, and it is just around the corner to break through solid tumors. Since tumor immunotherapy can cure tumors without the toxicity of traditional radiotherapy and chemotherapy, immune cell therapy is effective in tumors. has broad prospects for treatment. [0003] Adoptive cell therapy (ACT) has attracted much attention in clinical research due to its short-term expansion and activation of effector cells with antitumor activi...

AI Technical Summary

Problems solved by technology

It is difficult to introduce foreign genes into T lymphocytes by traditional direct transfection. Therefore, the lentiviral system used in the preparation of most CAR-T cells, because viral vectors involve many patent controversies, so research Researchers have also tried to prepare CAR-T cells by using transposons combined with electrotransduction to introduce foreign genes. Preclinical studies have also confirmed that this method is feasible, although retrovirus and lentivirus are considered safe for treatment with foreign genes. , but because the exogenous gene is randomly integrated into the host genome and exists in the cell for a long time and stably expressed, there are inappropriate insertion site mutations, which may cause the potential risk of cell transformation, and the way of transposon electroporation also exists with similar risks

Therefore, some studies have explored the direct introduction of coding plasmids into cells by electroporation, but because most of the plasmid introduction methods are transient expression, whether the therapeutic effect can be achieved needs further experimental verification

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