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Method for preparing human CIK (cytokine-induced killer) cell by combining novel TLR7 agonist GD

An agonist and cell technology, which is applied in the field of tumor immunotherapy and cellular immunology, can solve the problem of insignificant cytotoxic activity of heterogeneous cell populations, and achieve low preparation costs, increased tumoricidal activity, and easy large-scale production Effect

Inactive Publication Date: 2016-03-02
SHENZHEN HORNETCORN BIOTECH
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  • Claims
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AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to overcome the deficiencies of the prior art, on the basis of CIK cells optimized by the TLR7 small molecule agonist GD, to provide a preparation method and application of human CIK cells with a combination therapy concept, that is, GD-CIK , to solve the problem that the cytotoxic activity of heterogeneous cell populations does not increase significantly after CIK cell culture in the prior art; obtain anti-tumor adoptive immune cells with higher killing efficiency that can be used in clinical practice

Method used

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  • Method for preparing human CIK (cytokine-induced killer) cell by combining novel TLR7 agonist GD
  • Method for preparing human CIK (cytokine-induced killer) cell by combining novel TLR7 agonist GD
  • Method for preparing human CIK (cytokine-induced killer) cell by combining novel TLR7 agonist GD

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Embodiment 1

[0028] In Example 1, the obtained PBMCs were isolated and GD-CIK cells were cultured. Include the following steps:

[0029]1. Collect 25ml of peripheral venous blood from the patient, and obtain mononuclear cells by density gradient centrifugation of Ficoll-diatrizoate glucosamine. The specific steps are: 1500 rpm, centrifuge for 10 minutes, absorb the upper plasma layer, inactivate at 56°C for 30 minutes and centrifuge for later use, dilute the precipitated blood cells with normal saline, and divide human lymphocyte separation medium and diluted blood at a ratio of 1:2 Add the proportion into a centrifuge tube, centrifuge at 2000 rpm for 20 minutes, carefully absorb the buffy coat, wash twice with normal saline at 1600 rpm and 1300 rpm, and centrifuge for 7 minutes to obtain PBMCs .

[0030] 2. Induction culture of GD-CIK cells. The specific steps are: resuspend the suspension cells collected in Step 1 of Example 1 in GT-T551 containing 10% autoinactivated plasma TM In a ...

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Abstract

The invention discloses a method for preparing a high-toxicity CIK (cytokine-induced killer) cell by combining a novel TLR7 agonist GD. The method includes the steps of 1) collecting peripheral venous blood to obtain mononuclear cells; 2) adding the novel TLR7 agonist, a small molecule compound GD to perform CIK cell induced culture; 3) performing CIK cell multiplication culture to obtain a novel heterogeneous cell population, namely a GD-CIK cell. The method has the advantages that compared with a CIK cell cultured by a conventional method, the GD-CIK cell prepared by the method is obviously higher in tumor killing cytotoxic activity and is up to above 95% in tumor killing rate (effector-target ratio: 20:1); the method is low in preparation cost, simple in process, easily controllable in condition, low in requirement on equipment, easy for scale production and the like; the GD-CIK cell prepared by the method is mainly applied to cancer patient treatment or prevention of high-risk groups of cancers.

Description

technical field [0001] The invention belongs to the fields of cellular immunology and tumor immunotherapy, and relates to a method for preparing human CIK cells in combination with a novel TLR7 agonist GD and the application of anti-tumor adoptive immunotherapy. Background technique [0002] In recent years, with the change of lifestyle, the deterioration of the natural environment, and the improvement of medical technology, the number of tumors detected is on the rise, and tumors have become one of the important diseases that seriously threaten human health. Due to the high damage and high mortality rate of tumors, the research and application of anti-tumor treatments have always been the focus of attention. Tumor immunotherapy is the fourth largest therapy for clinical tumor treatment, among which immune cell therapy It has been widely used in China. At present, adoptive reinfusion therapy of immune cells is the most widely used in China. [0003] CIK cells, cytokine-indu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
Inventor 高东李旺陈艳媛王宇环魏志璋魏宗科罗晓玲
Owner SHENZHEN HORNETCORN BIOTECH
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