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Chimeric antigen receptor T cell and application thereof

A technology of chimeric antigen receptors and cells, applied in receptors/cell surface antigens/cell surface determinants, applications, animal cells, etc., can solve the problem of not being able to completely cure tumor patients, and alleviate the problem of drug resistance , Improve the effect of anti-tumor effect

Pending Publication Date: 2020-07-14
格源致善(上海)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The efficacy of PD-L1 monoclonal antibody alone is not enough to completely cure tumor patients, and many clinical phase III trials failed because they did not reach the ultimate goal

Method used

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  • Chimeric antigen receptor T cell and application thereof
  • Chimeric antigen receptor T cell and application thereof
  • Chimeric antigen receptor T cell and application thereof

Examples

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Effect test

Embodiment 1

[0069] This embodiment provides a chimeric antigen receptor T cell expressing both the CD19 CAR structure and the PD-L1 CAR structure.

[0070] The preparation method of the chimeric antigen receptor T cell comprises the following steps:

[0071] 1. Link the nucleotide sequence (ie, SEQ ID NO.10 and SEQ ID NO.11) expressing the chimeric receptor targeting CD19 CAR and PD-L1 CAR with the nucleotide sequence encoding 2A to synthesize a fusion gene , when synthesizing this gene, the restriction endonucleases HindIII and BamHI restriction endonuclease sites were included at both ends, and loaded onto the pDNA3.1(+) vector; the sequence was entrusted to Shanghai Sangon Biotechnology Co., Ltd. to synthesize.

[0072] 2. Use HindIII and BamHI to double digest the pDNA3.1(+) vector containing the target gene, and then cut the gel to recover the target gene fragment;

[0073] 3. Digest the original vector pCDH-EF1-Luc2-T2A-tdTomato with HindIII and BamHI, and recover a vector fragment...

Embodiment 2

[0077] The difference with Example 1 is that SEQ ID NO.11 is replaced by SEQ ID NO.12, the preparation method is the same as in Example 1, and the expressed amino acid sequences are SEQ ID NO.3, SEQ ID NO.9 and SEQ ID NO.9. Amino acid sequence composed of ID NO.7.

Embodiment 3

[0079] The difference with Example 1 is that SEQ ID NO.11 is replaced by SEQ ID NO.13, its preparation method is the same as in Example 1, and the amino acid sequences expressed are SEQ ID NO.3, SEQ ID NO.9 and SEQ ID NO.9 and SEQ ID NO.13. Amino acid sequence composed of ID NO.8.

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Abstract

The invention provides a chimeric antigen receptor T cell and application thereof. The chimeric antigen receptor T cell expresses chimeric antigen receptors targeting CD19 and PD-L1 at the same time.The chimeric antigen receptor T cell expresses a CD19 CAR structure and a PD-L1 CAR structure at the same time, and after PD-L1 CAR is combined with PD-L1 expressed by a tumor cell, an activation signal can be transmitted intracellularly. Therefore, the chimeric antigen receptor T cell not only can recognize the tumor cells expressing CD19 in a targeted manner, but also can relieve immunosuppression of PD-1 expressed by the tumor cells on the T cells, avoid an immune escape mechanism of the tumor cells and relieve the problem of drug resistance of CAR-T treatment; and a new idea is provided for CAR-T treatment and adoptive feedback immune cell treatment.

Description

technical field [0001] The invention belongs to the field of tumor cell immunotherapy, and in particular relates to a chimeric antigen receptor T cell and its application, in particular to a T cell expressing two chimeric antigen receptors simultaneously and its application. Background technique [0002] Adoptive Cell Transfer Therapy (ACT) refers to the isolation of immunocompetent cells from tumor patients, expansion and functional identification in vitro, and then reinfusion to patients, so as to directly kill tumors or stimulate the body's immune response The purpose of killing tumor cells. In recent years, Chimeric Antigenreceptor (CAR) modified T lymphocyte (CAR-modified T lymphocyte, CAR-T) therapy has achieved remarkable efficacy in the treatment of hematological malignancies, especially for acute lymphoblastic leukemia. (Acute lymphoblasticleukemia, ALL) and B-cell lymphoma, the remission rate reaches about 80%. [0003] Among them, the key to CAR-T design is to s...

Claims

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Application Information

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IPC IPC(8): C12N5/10C07K19/00C12N15/62C12N15/867A61K39/00A61P35/00A61P35/02
CPCC12N5/0636C07K16/2803C07K16/2827C07K14/7051C12N15/86A61K39/001112A61K39/001111A61P35/00A61P35/02C12N2510/00C07K2319/03C07K2319/33C07K2319/74C12N2740/15043C12N2800/107A61K2039/5156A61K2039/804
Inventor 苏小平李锐
Owner 格源致善(上海)生物科技有限公司
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