TCR knockout T cell receptor gene-modified T cell targeting TIPE3, preparation method and application thereof

A cell receptor and gene modification technology, applied in the field of medical biology, can solve problems such as the loss of target recognition ability, and achieve the effect of avoiding autoimmune reactions and preventing mismatches

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)
A...

Method used

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  • TCR knockout T cell receptor gene-modified T cell targeting TIPE3, preparation method and application thereof
  • TCR knockout T cell receptor gene-modified T cell targeting TIPE3, preparation method and application thereof
  • TCR knockout T cell receptor gene-modified T cell targeting TIPE3, preparation method and application thereof

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Embodiment 1

[0057] A method for preparing TCR knockout T cell receptor gene-modified T cells targeting TIPE3, comprising the following steps:

[0058] (1) Preparation of TCR-knockout CD3-positive T lymphocytes

[0059] a) Isolation of PBMC (peripheral blood mononuclear cells)

[0060] 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.

[0061] 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.

[0062] b) Isolation of antigen-specific T lymphocytes by immunomagnetic bead method

[0063] Take the above PBMC, add serum-free basal medium to make a cell suspension; add CD3 / CD...

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Abstract

The invention provides a method for preparing TCR knockout T cell receptor gene-modified T cells targeting TIPE3, which comprises the following steps: by knocking out the TCR gene on T lymphocytes andpreparing T cell receptors targeting TIPE3, and the TCR knockout T cell receptor gene-modified T cell targeting TIPE3 is obtained. The TCR knockout T cell receptor gene-modified T cell targeting TIPE3 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 killing performanceof TIPE3 positive tumor cells can be kept. The method also provides an application of TCR knockout T cell receptor gene-modified T cells targeting TIPE3.

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 TIPE3, a preparation method and application thereof. Background technique [0002] Tumor necrosis factor α-inducible protein 8-like 3 (TNFAIP8L3, TIPE3) is a member of the TNFAIP3 family, which acts as a lipid second messenger transport protein to promote carcinogenesis, and acts as a feedback enhancer to regulate carcinogenesis and phospholipid pathways. TIPE3mRNA can be detected in many tissues, but TIPE3 protein is rarely expressed in normal tissues, but its expression is significantly increased in cancerous tissues. Therefore, TIPE3 is a very suitable tumor-specific target for cellular immunotherapy. [0003] In recent years, immune cell therapy has become a treatment method with great prospects in the treatment of malignant tumors after surgery, radiotherapy, and chemotherapy. T cell receptor (T cell receptor, TCR...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/113C12N15/867C12N7/01A61P35/00
Inventor 曾滢张长风龙丽梅
Owner SHENZHEN BINDEBIOTECH CO LTD
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