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Immune cell and preparation and application thereof

A technology of immune cells and preparations, applied in the field of preparations and applications of immune cells, which can solve the problems of short survival time, low lethality, and less effective effect than hematological tumors, and achieve the effect of lasting time and reducing exhaustion

Active Publication Date: 2020-10-30
GUANGDONG XIANKANGDA BIOTECH CO LTD
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are still some problems in CAR-T therapy. The effect of CAR-T cells in treating solid tumors has not been as good as that of hematological tumors. One of the important reasons is that the exhaustion of CAR-T cells leads to short survival time in vivo and low lethality.

Method used

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  • Immune cell and preparation and application thereof
  • Immune cell and preparation and application thereof
  • Immune cell and preparation and application thereof

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Experimental program
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Effect test

Embodiment 1

[0093] The preparation method and functional verification of immune cells in this embodiment include the following steps:

[0094] Step 1: Isolation of peripheral blood PBMCs and expansion of CAR-T cells

[0095] 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 a 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 Cultured in a constant temperature incubator at 37°C with 5% CO2.

[0096] Step 2: Cell Line Culture

[0097] Cell line expressing GPC3: Huh-7 (human liver cancer cell), purchased from ATCC.

[0098] Cell line not expressing GPC3: A549 (human non-small cell lung cancer cells), purchased from ATCC.

[0099] Cells...

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Abstract

The invention relates to the technical field of biological cells, in particular to an immune cell and a preparation and application thereof. The expression of the CAR is regulated and controlled to depend on a segment of amino acid sequence. The invention provides a design scheme of an adjustable CAR and an engineered immune cell. The CAR designed by the design scheme is transferred into T cells;when T cells meet target cells, the expression of the CAR rises, tumor surface markers are recognized, the driving killing function is achieved, and when no target cells exist, the expression of the CAR is automatically lowered so that the exhaustion of the CAR-T cells is reduced, and the driving function of the CAR-T cells in vivo lasts for a longer time.

Description

technical field [0001] The present invention relates to the technical field of biological cells, in particular to immune cells that can regulate the expression of CAR, optimize the design of CAR, and the preparation and application of the immune cells. Background technique [0002] In 2012, the annual meeting of the International Society for Cell Therapy pointed out that biological immune cell therapy has become the fourth means of tumor treatment besides surgery, radiotherapy, and chemotherapy, and will become a must-have means for future tumor treatment. Immune cell therapy is to collect peripheral venous blood from patients, isolate peripheral blood mononuclear cells in a GMP laboratory, and induce a large number of immune effector cells with high anti-tumor activity under the induction of various cytokines, and then through vein, intradermal Injection, intervention, etc. are reinfused into the patient's body to achieve the purpose of enhancing the patient's immune functi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C07K19/00A61K39/00A61P35/00A61P35/02
CPCA61K39/0011A61P35/00A61P35/02C07K14/7051C07K14/7155C07K2319/02C07K2319/03C07K2319/33C12N5/0636C12N5/0638C12N5/0646C12N2510/00
Inventor 谢海涛薛卫巍马丽雅
Owner GUANGDONG XIANKANGDA BIOTECH CO LTD
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