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Preparation method of tumor fusion cell vaccine

A technology for fused cells and cells, applied in fused cells, biochemical equipment and methods, antineoplastic drugs, etc.

Inactive Publication Date: 2016-11-09
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the defects of current anti-tumor immunotherapy, the present invention provides a method for preparing a tumor fusion cell vaccine, which is a method for preparing a vaccine by using αβTCR gene to transfect γδT cells and tumor cells to form fusion cells

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  • Preparation method of tumor fusion cell vaccine
  • Preparation method of tumor fusion cell vaccine
  • Preparation method of tumor fusion cell vaccine

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Embodiment 1

[0020] Example 1 A preparation method of tumor fusion cell vaccine

[0021] 1. Isolation of Peripheral Blood Mononuclear Cells (PBMCs)

[0022] Take PBMC from peripheral blood mononuclear cells (or dilute peripheral blood samples 1:1 with RPMI 1640 medium, add to a centrifuge tube containing lymphocyte separation medium (volume ratio of diluted venous blood to lymphocyte separation medium 2:1), 500×g Centrifuge for 20min. Aspirate the milky white mononuclear cell layer at the interface of the lymphocyte separation medium, wash twice with RPMI 1640 medium, and adjust the cell concentration to 10 with complete medium. 6 cells / mL for later use.

[0023] 2. Preparation of γδ T cells

[0024] 1) Separation of γδT cells from normal people and patients with OS, count the total amount of peripheral blood mononuclear cells, adjust the cell concentration to 1-2×106 / mL, take a 24-well plate, add 1 mL of cell suspension to each well, and place at 37°C , 5%CO 2 Culture in the incubator...

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Abstract

The invention provides a preparation method of a tumor fusion cell vaccine. The preparation method comprises the following steps of fusing genes of an alpha-TCR (T cell receptor) and a beta-TCR into an alpha and beta-TCR; obtaining a recombinant plasmid; constructing the alpha and beta-TCR into a shuttle plasmid pMP71-alpha and beta-TCR; transfecting gamma-delta T cell by the gene of alpha and beta-TCR; using an electric fusion technique to fuse the gamma-delta T cell and tumor cell into a vaccine. The preparation method has the advantages that the fusion cell is formed by the gamma-delta T cell transfected by the gene of alpha and beta-TCR and the tumor cell, so as to prepare the vaccine; the specificity of tumor cell can be identified, the capability of the fusion cell inducing the withering of the tumor cell is strongest, and the active immunity of the body receptor is stimulated to treat tumors.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a vaccine preparation method, in particular to a vaccine preparation method for tumor fusion cells, which is a method for preparing a vaccine by transfecting γδ T cells with tumor cells to form fusion cells with αβTCR genes. Background technique [0002] Malignant tumor is a disease that seriously endangers human health, and its incidence is increasing year by year. According to data from the Cancer Prevention and Control Office of the Ministry of Health, in the past 20 years, one out of every four to five deaths in China has died of cancer, which has become the number one killer threatening national health. There are about 2.6 million cancer cases and 1.8 million deaths in my country every year. [0003] In recent years, tumor immunotherapy has attracted much attention and has become the fourth treatment method after surgery, radiotherapy and chemotherapy. There are mainly two metho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61P35/00C12N5/10C12N5/22
CPCA61K39/0011A61K2039/5152C07K14/7051C12N5/163
Inventor 朱健陶惠民梁成振
Owner ZHEJIANG UNIV
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