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Method for detecting anti-CD19 chimeric antigen receptor T cell effects of inhibiting leukemia cells

A technology of chimeric antigen receptors and leukemia cells, which is applied in the direction of receptors/cell surface antigens/cell surface determinants, cells modified by introducing foreign genetic materials, chemical instruments and methods, etc., can solve the problems of industrialization, Can not guarantee the accuracy of the inhibitory effect of CART cells on tumor cells and other issues, to achieve high accuracy, easy operation, and short time-consuming effects

Active Publication Date: 2015-12-16
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Abstract
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Problems solved by technology

[0005] Aiming at the defects in the prior art that the accuracy of verifying the inhibitory effect of CART cells on tumor cells cannot be guaranteed, and the industrialization cannot be realized, the present invention provides a method for inhibiting the inhibitory effect of CD19 chimeric antigen receptor T cells on leukemia cells. The purpose of realization is to obtain a verification method that is safe, reliable, high-precision and can be applied industrially

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  • Method for detecting anti-CD19 chimeric antigen receptor T cell effects of inhibiting leukemia cells
  • Method for detecting anti-CD19 chimeric antigen receptor T cell effects of inhibiting leukemia cells
  • Method for detecting anti-CD19 chimeric antigen receptor T cell effects of inhibiting leukemia cells

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Embodiment 1

[0026] Embodiment 1: A method for detecting the inhibitory effect of anti-CD19 chimeric antigen receptor T cells on leukemia cells disclosed in the present invention, taking B-cell leukemia cells as an example, the detection method includes the following steps, (1) expressing anti-CD19 Chimeric antigen receptor T cells and B cell leukemia cells were mixed in equal proportions and then transplanted into the immune deficiency mouse model: equal proportions of cells generally refer to equal proportions of the number of cells, such as anti-CD19 chimeric antigen receptor T 5x10 each for cells and B-cell leukemia cells 6 A cell, the anti-CD19 chimeric antigen receptor T cell preservation number CCTCCNO:C201503, the cell is preserved in the China Center for Type Culture Collection (Wuhan), the address is Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province (opposite to the No. 1 Primary School Attached to Wuhan University), Wuhan University Preservation ...

Embodiment 2

[0030] Embodiment 2: A method for detecting the inhibitory effect of anti-CD19 chimeric antigen receptor T cells on leukemia cells disclosed in the present invention, taking B-cell leukemia cells as an example, the detection method includes the following steps, (1) expressing anti-CD19 Chimeric antigen receptor T cells and B cell leukemia cells were mixed in equal proportions and then transplanted into the immune deficiency mouse model: equal proportions of cells generally refer to equal proportions of the number of cells, such as anti-CD19 chimeric antigen receptor T 5x10 each for cells and B-cell leukemia cells 6 1 cell, anti-CD19 Chimeric Antigen Receptor T cell with the preservation number CCTCCNO:C201503, preserved in the China Center for Type Culture Collection (Wuhan), and the address is Wuhan University Campus (Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province) Opposite to the First Affiliated Primary School of the University), Wuhan Un...

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Abstract

The invention discloses a method for detecting anti-CD19 chimeric antigen receptor T cell effects of inhibiting leukemia cells. The method comprises 1, mixing anti-CD19 chimeric antigen receptor T cells and leukemia cells according to a certain ratio, and transplanting the mixed cells into an immunodeficient mouse model, wherein the anti-CD19 chimeric antigen receptor T cell has an accession number of CCTCC NO: C201503, 2, carrying out immunodeficient mouse model peripheral blood cell dyeing, and 3, detecting the dyed peripheral blood cells by a flow cytometer. Human leukemia cells and leukemia cell inhibition cells are transplanted into an immunodeficient mouse model together so that effects of inhibiting different human leukemia cells can be really shown and high effectiveness and accuracy of the inhibition effect detection method are guaranteed.

Description

technical field [0001] The invention belongs to the technical field of biological chimeric antigen receptor T cells, in particular to a method for detecting the inhibitory effect of anti-CD19 chimeric antigen receptor T cells on leukemia cells. Background technique [0002] Chimeric antigen receptor T cells (CART cells) are T cells that express a chimeric receptor on their surface that recognizes a specific antigen and can transmit signals. T cells play an important role in anti-tumor. CART cells are T cells that express such a molecule: the extracellular segment is the variable region of the antibody heavy chain and light chain connected by a peptide to form a single-chain variable region (ScFv ), the intracellular segment is a chimera of various signaling molecules, including CD3zeta, CD28, OX-40, 4-1BB, etc., and the transmembrane region comes from other molecules (such as CD8, CD4, CD28 and CD3zeta) transmembrane region. CART cells recognize specific molecules on the s...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/50C12N15/867C12N15/13C12N5/10
CPCC07K14/705C07K16/2803C07K2319/30C12N15/86C12N2740/15041G01N33/505G01N33/57426G01N2333/7051G01N2800/52
Inventor 李鹏赖允鑫林思妙姚瑶秦乐魏新茹李伯衡
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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