Immune cell for expressing cytokine receptor fusion type chimeric antigen receptor and application thereof

A chimeric antigen receptor and cytokine technology, applied in the field of biological cells, can solve the problems of solid tumors without good treatment effect, off-target, low remission rate, etc., to reduce the influence of tumor microenvironment, inhibit growth, and enhance proliferation ability. Effect

Inactive Publication Date: 2021-09-21
SHENZHEN FIRST CONDOR BIOSCIENCE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although CAR-T cell therapy shines in the treatment of hematological tumors, it does not have a good therapeutic effect on solid tumors, with a low response rate and prone to off-target and other toxic side effects

Method used

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  • Immune cell for expressing cytokine receptor fusion type chimeric antigen receptor and application thereof
  • Immune cell for expressing cytokine receptor fusion type chimeric antigen receptor and application thereof
  • Immune cell for expressing cytokine receptor fusion type chimeric antigen receptor and application thereof

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Experimental program
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Effect test

Embodiment 1

[0084] The preparation method and functional verification of the immune cells in this embodiment include the following steps:

[0085] S100: Isolation of peripheral blood PBMCs and culture of T cells.

[0086] Mononuclear cells were isolated from donor peripheral blood, density gradient centrifugation was performed using ficol, and T cells were enriched with a T cell sorting kit (CD3 MicroBeads, human-lyophilized, 130-097-043), using conjugated anti-CD3 / anti-CD28 magnetic beads activated culture and expansion of T cells; the medium used TexMACS GMP Medium (MiltenyiBiotec, 170-076-309), containing 10% FBS, 2mM L-glutamine, 100IU / ml rhIL2, all cells were placed At 37°C, 5% CO 2 Cultured in a constant temperature incubator to obtain T cells.

[0087] S200: Cell line culture.

[0088] Cell line not expressing Claudin18.2: 293T (human embryonic kidney cell line), purchased from ATCC.

[0089] Cell line expressing Claudin18.2: 293T-CLDN18.2, constructed by self-produced lentiviru...

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Abstract

The invention discloses an immune cell for expressing a cytokine receptor fusion type chimeric antigen receptor and application of the immune cell. A cytokine receptor fusion type chimeric antigen receptor is expressed on a cell membrane of the immune cell, and a cytokine receptor intracellular domain is fused in an intracellular domain structure of the cytokine receptor fusion type chimeric antigen receptor. The immune cell can enhance the proliferation capacity of the cell, reduce the influence of a tumor microenvironment and enhance the tumor killing activity of the immune cell under the condition of receiving antigen stimulation, so that the negative regulation of the tumor microenvironment on the immune cell is weakened, the exhaustion of the immune cell is reduced, and corresponding JAK and STAT signal channels are activated by combining with a cytokine receptor, the cells are induced to secrete various active substances such as IFN-gamma, perforin and granzyme, so that the growth of tumor cells is inhibited, the anti-tumor capability is enhanced, and the inhibition of a tumor microenvironment on immune cells is reduced.

Description

technical field [0001] The invention relates to the technical field of biological cells, in particular to an immune cell expressing cytokine receptor fusion type chimeric antigen receptor and its application. Background technique [0002] In recent years, tumor immunotherapy has developed rapidly, especially adoptive cell therapy (ACT), which means that immune cells such as T cells and NK cells are isolated from patients, modified and expanded in vitro, and then reintroduced into patients for tumor treatment. method of treatment. In 2013, the immunotherapy of tumors was rated as the first among the "Top Ten Breakthroughs" by Science Impurities. [0003] CAR-T and TCR-T are important components of adoptive cell therapy, especially CAR-T therapy, which has made remarkable achievements in the treatment of blood tumors and achieved a high remission rate. A typical CAR structure consists of three parts Composition, extracellular recognition of tumor antigen scFv, hinge and tran...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/62A61K35/17A61P35/00
CPCC07K14/7051C07K14/715A61K35/17A61P35/00C07K2319/02C07K2319/03
Inventor 都晓龙马丽雅方晓
Owner SHENZHEN FIRST CONDOR BIOSCIENCE CO LTD
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