TCR knockout T cell receptor gene-modified T cell targeting NY-ESO-1, preparation method and application thereof

A TCR-NY-ESO-1, cell receptor technology, applied in the field of medical biology, can solve the problem of loss of target recognition ability

Inactive Publication Date: 2019-06-04
SHENZHEN BINDEBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, studies have shown that there is a mismatch between the TCR transduced by TCR-T technology and the endogenous TCR subunits (such as α chain and β chain) of T cells, resulting in the target recognition ability of the mismatched TCR. Autoimmune response caused by loss or recognition of self-antigen or major histocompatibility complex (MHC)
At the same time, it is still a huge challenge to obtain TCR genes with high specificity and high affinity for tumor antigens and the correct pairing of TCR subunits
At present, there is no research and research on targeting the T cell receptor of NY-ESO-1 while avoiding the mismatch between TCR transduced by TCR-T technology and T cell endogenous TCR subunits

Method used

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  • TCR knockout T cell receptor gene-modified T cell targeting NY-ESO-1, preparation method and application thereof
  • TCR knockout T cell receptor gene-modified T cell targeting NY-ESO-1, preparation method and application thereof
  • TCR knockout T cell receptor gene-modified T cell targeting NY-ESO-1, preparation method and application thereof

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Embodiment 1

[0058] A method for preparing T cell receptor gene-modified T cells targeting NY-ESO-1 knocked out by TCR, comprising the following steps:

[0059] (1) Preparation of TCR-knockout CD3-positive T lymphocytes

[0060] a) Isolation of PBMC (peripheral blood mononuclear cells)

[0061] PBMCs are derived from autologous venous blood, autologous bone marrow, umbilical cord blood, and placental blood. It is best to come from fresh peripheral blood or bone marrow collected from cancer patients one month after surgery and one month after radiotherapy and chemotherapy.

[0062] The patient's blood was drawn and sent to the blood separation chamber; peripheral blood mononuclear cells were collected, and the middle layer cells were collected after Ficoll centrifugation; after washing with PBS, PBMCs were obtained.

[0063] b) Isolation of antigen-specific T lymphocytes by immunomagnetic bead method

[0064] Take the above PBMC, add serum-free basal medium to make a cell suspension; add...

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Abstract

The invention provides a method for preparing TCR knockout T cell receptor gene-modified T cells targeting NY-ESO-1, which comprises the following steps: by knocking out the TCR gene on T lymphocytesand preparing T cell receptors targeting NY-ESO-1, and the TCR knockout T cell receptor gene-modified T cell targeting NY-ESO-1 is obtained. The TCR knockout T cell receptor gene-modified T cell targeting NY-ESO-1 can effectively prevent the mismatch of endogenous and exogenous TCR alpha-chain and beta-chain existing in T cells, autoimmune responses are avoided, and efficient and specific killingperformance of NY-ESO-1 positive tumor cells can be kept. The method also provides an application of the TCR knockout T cell receptor gene-modified T cells targeting NY-ESO-1.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to a TCR knockout T cell receptor gene-modified T cell targeting NY-ESO-1, a preparation method and application thereof. Background technique [0002] Among many tumor antigen families, cancer-testis antigen (CTA) has strong specificity and is widely expressed in tumor tissues. The expression levels of CTA are different in different stages and different differentiation levels of tumors, which may be related to the deterioration and progression of tumors. Among them, NY-ESO-1 (New York esophageal squamous cell carcimonma 1) belongs to the CTA family and is considered to be one of the most immunogenic tumor antigens. It is expressed in ovarian cancer, etc., but it is almost not expressed in other normal cells, so it has attracted attention. [0003] In recent years, immune cell therapy has become a treatment method with great prospects in the treatment of malignant tumors after surger...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/113C12N15/867C12N7/01A61P35/00
Inventor 张长风吴咏东
Owner SHENZHEN BINDEBIOTECH CO LTD
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