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Preparation method of polylineage activated killer cells for tumor immunotherapy

An immunotherapy and cell technology, applied in the direction of blood/immune system cells, antineoplastic drugs, animal cells, etc., can solve the problems of poor curative effect and applicability

Inactive Publication Date: 2014-12-24
昆明市第一人民医院
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Problems solved by technology

A very important characteristic of tumors is their "heterogeneity". Therefore, it is difficult to completely eliminate tumor cells by using only one effector cell with a specific phenotype, and its efficacy and applicability are poor.

Method used

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  • Preparation method of polylineage activated killer cells for tumor immunotherapy
  • Preparation method of polylineage activated killer cells for tumor immunotherapy
  • Preparation method of polylineage activated killer cells for tumor immunotherapy

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Experimental program
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Embodiment Construction

[0024] A method for preparing multiline activated killer cells for tumor immunotherapy, specifically:

[0025] Step 1. Isolate mononuclear cells from the peripheral blood of patients with malignant tumors or healthy people:

[0026] Peripheral blood mononuclear cell isolation: 50 mL of human peripheral blood was collected under sterile conditions. A 50mL syringe was used for blood collection, and heparin was used as an anticoagulant at a concentration of 15 IU heparin per milliliter of peripheral blood, and the volume ratio of heparin solution to blood was 1:9. Inject anticoagulant blood into a 50mL centrifuge tube and centrifuge at 400g for 10 minutes; carefully draw the upper layer of plasma into a 50mL centrifuge tube, add the remaining anticoagulant blood to normal saline at a ratio of 1:1 and mix well; take a 50mL centrifuge tube and add human peripheral blood Lymphocyte separation medium (Tianjin Haoyang), carefully add diluted peripheral blood to the upper layer of lym...

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Abstract

The invention discloses a preparation method of polylineage activated killer (PAK) cells for tumor immunotherapy. The method comprises the following steps: separating mononuclear cells from peripheral blood of patients with malignant tumor or healthy persons, adding the mononuclear cells of the peripheral blood into anti-TCR gamma delta monoclonal antibodies and IFN-gamma in-vitro induced PAK cells, carrying out in-vitro induction on the obtained objects for 72 hours, and carrying out amplification on the PAK cells in the presence of anti-TCR gamma delta monoclonal antibodies, anti-CD3 monoclonal antibodies and IL-2. The prepared PAK cells are necessary to contain TCR gamma delta<+> cells, CD3<->CD56<+> (NK) cells, CD3<+>CD56<+>(NKT) cells and CD3<+>CD8<+>cells, therefore, the prepared PAK cells have a stronger antitumor effect, and the killing activity of the prepared PAK cells on human lung adenocarcinoma A549 and human lung squamous carcinoma YTMLC is higher than that of cytokine induced killer (cytokine induced killer CIK) cells.

Description

technical field [0001] The invention relates to a treatment method for human tumor diseases, in particular to a preparation method for polylineage activated killer (PAK) cells specially used for tumor immunotherapy. Background technique [0002] Compared with traditional tumor treatment methods, adoptive cell therapy (ACT), which reinfuses immune cells back into the body, can directly kill tumor cells without damaging the structure and function of the immune system; at the same time, it can regulate and strengthen the immune system. Therefore, it has become an important adjuvant therapy for tumor surgery, radiotherapy and chemotherapy, and provides a new way to prevent tumor recurrence and metastasis and improve the quality of life of patients with advanced tumors. ACT has achieved certain curative effect in the treatment of malignant melanoma, renal cancer, lymphoma and other tumors. [0003] Human peripheral blood mononuclear cells (PBMC) contain a variety of immune effec...

Claims

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Application Information

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IPC IPC(8): C12N5/0786A61P35/00
Inventor 苏晓三汪珺王翼寅陈睿杨柳张蕾
Owner 昆明市第一人民医院
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