Combined factor for inducing tumor specific T cells and method for obtaining tumor specific T cells

A tumor-specific and cytokine technology, which is applied in the field of combining factors for inducing tumor-specific T cells and obtaining tumor-specific T cells, can solve problems such as time-consuming, limited wide application, and difficulty in obtaining TIL cells

Inactive Publication Date: 2016-02-24
SYZ CELL THERAPY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the induction and culture of tumor-specific CTLs takes a lot of time and requires high technical requirements, which greatly limits the wide application of this treatment method in cancer patients.
In early animal experiments, some scholars found that the addition of IL-2 to the culture system could amplify cells with tumor cytotoxicity, but the research found that the method was not strong enough to induce cell lethality, and the efficiency of inducing specific CTL was low.
Subsequently, Professor Rosenberg and others developed TIL technology research, but the difficulty of obtaining TIL cells limited its clinical application

Method used

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  • Combined factor for inducing tumor specific T cells and method for obtaining tumor specific T cells
  • Combined factor for inducing tumor specific T cells and method for obtaining tumor specific T cells
  • Combined factor for inducing tumor specific T cells and method for obtaining tumor specific T cells

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Example 1: Example of tumor-specific T cell induced enrichment isolation

[0037] 1. Induced enrichment and isolation of tumor-specific T cells

[0038] 1. 2 patients with stage B liver cancer were informed and signed the informed consent, and 50ml of peripheral venous blood was drawn with heparin sodium anticoagulant tube respectively. Lymphocyte separation medium (FreseniusKabiNorgeAS, LymphoprepTM) was used to separate PBMCs, and they were suspended with fresh serum-free medium (AIM-V from LifeTechnology), and the cell density was 2×10 6 / ml. In each milliliter of serum-free medium, add liver cancer antigen peptide 2μg / ml; IL-250U / ml; IL-7 and IL-15 were respectively 5ng / ml. 37°C, 5% CO 2 Cultivated in an incubator. The core epitope of the liver cancer antigen library is FLPSDFFPS (SEQ ID NO: 1), which was synthesized by Shanghai Chupeptide Biotechnology Co., Ltd.

[0039] 1.2 After 72 hours of culture, IFNγ enrichment sorting (Miltenyi Biotec 130-054-201) was p...

Embodiment 2

[0051] Example 2: Cervical cancer-specific T cell induced enrichment and isolation test

[0052] 1. Induced enrichment and isolation of tumor-specific T cells

[0053] 1.1 One patient with cervical cancer was informed and signed the informed consent, and 50ml of peripheral venous blood was drawn with heparin sodium anticoagulation tube. Lymphocyte separation medium (FreseniusKabiNorgeAS, LymphoprepTM) was used to separate PBMCs, and they were suspended with fresh serum-free medium (AIM-V from LifeTechnology), and the cell density was 2×10 6 / ml. In each milliliter of serum-free culture medium, add cervical cancer antigen peptide 2μg / ml; IL-250U / ml; IL-7 and IL-15 were respectively 5ng / ml. Incubate in a 37°C, 5% CO2 incubator. Among them, the cervical cancer antigen peptide library, the core antigen epitope of which is KLPDLCTEL (SEQ ID NO: 2), was synthesized by Shanghai Chupeptide Biotechnology Co., Ltd.

[0054] 1.2 After 72 hours of culture, IFNγ enrichment sorting (Milte...

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Abstract

The invention discloses a combined factor for inducing tumor specific T cells and a method for obtaining the tumor specific T cells. The combined factor comprises a tumor specific antigen and cell factors IL-2, IL-7 and IL-15. The method comprises the following steps: suspending peripheral blood PBMC with a fresh serum-free lymphocyte culture solution, wherein the cell density is 2*10<6>-5*10<6> / ml; adding a stimulus of the combined factor for inducing the tumor specific T cells, conducting culturing in a 5% CO2 culturing box at 37 DEG C for 2-10 days, and separating the IFN gamma positive antigen specific T cells through an IFN gamma magnetic bead enrichment method. The method can be used for obtaining the high-purity tumor antigen specific T cells, and is simple, and low in cost. The prepared tumor antigen specific T cells can be applied to preparation of medicines or vaccines for clinical cell treatment and tumor immunization treatment.

Description

technical field [0001] The invention belongs to the field of in vitro induction, separation and culture of immune cells, and relates to a combination factor for inducing tumor-specific T cells and a method for obtaining tumor-specific T cells. Background technique [0002] Malignant tumors seriously endanger human life and health, and its mortality rate ranks first among various diseases. According to estimates by the World Health Organization (WHO) and the American Cancer Society, about 12 million new cancer patients are newly diagnosed worldwide each year, and about 8 million cancer patients die (20,000 cancer patients die every day). There are currently 10 million cancer patients in China. Common malignant tumors such as liver cancer, lung cancer, breast cancer and prostate cancer have increased significantly in recent years, and they are becoming younger, posing a serious threat to human health. [0003] The three existing conventional tumor treatment methods (ie surger...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783A61K35/17A61P35/00
Inventor 陶然韩研妍李进周向军
Owner SYZ CELL THERAPY
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