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Preparation of specific tumor killing cell

A cell-killing and species-specific technology, applied in the field of anti-tumor cellular immunotherapy, can solve the problems of low clinical effective rate and low tumor killing rate

Active Publication Date: 2012-07-04
玥特农生物科技河北有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] The main purpose of the present invention is to solve the bottleneck problem of low tumor killing rate and low clinical effective rate of the current DC-CIK method, apply this technology, can significantly improve the killing efficiency of tumors, improve clinical effective rate and objective curative effect

Method used

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  • Preparation of specific tumor killing cell
  • Preparation of specific tumor killing cell
  • Preparation of specific tumor killing cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Use the COBE SPECTRA cell component separator, set the parameters and program according to the instruction manual, collect 6000-8000ml of peripheral blood, and collect 120-140ml of mononuclear cells. Divide into 50ml centrifuge tubes, and centrifuge at 1500 rpm for 5 minutes in a Hitachi cell centrifuge. Collect the upper layer of plasma and transfer it to a 50ml centrifuge tube to prepare autologous inactivated serum (method: add 2.5ml of 10% calcium gluconate by pipetting and mixing, bathe in 56°C water for 35 minutes, centrifuge at 2500 rpm for 5 minutes, collect the supernatant), and refrigerate at 4°C spare. Collect the cells, dilute them with 30ml of normal saline, transfer them into a 50ml centrifuge tube containing 15ml of lymphocyte separation medium (Ficoll, 1.077), centrifuge at 2000rpm for 15 minutes in a horizontal rotor centrifuge, and absorb the middle white cell layer (monocytes) with a disposable pipette. ). Take 15ml of mononuclear cells and transfer...

Embodiment 2

[0110] Example 3

[0111] The present invention also includes a combined reagent, which is characterized in that it includes all the reagents needed to prepare specific DC-CIK-CTL cells and preparations thereof, and the names of the reagents are as follows:

Embodiment 3

[0113] 11. Monocyte macrophage colony stimulating factor (GM-CSF);

[0114] 12. Interleukin 4 (IL-4);

[0115] 13. Gentamicin;

[0116] 14. Autologous tumor-associated whole antigen;

[0117] 15. IFN-γ;

[0118] 16. CD3 monoclonal antibody;

[0119] 17. Interleukin-2 (IL-2);

[0120] 18. 0.9% sodium chloride;

[0121] 19. Human albumin;

[0122] 20. Interleukin-2 (IL-2) for injection;

[0123] The doses of the reagents are as follows:

[0124]Doses and final concentrations of DC-CIK-CTL combination reagents are as follows:

[0125] 11. GM-CSF: 50ug / 500ml;

[0126] 12. IL-4: 30ug / 500ml;

[0127] 13. Gentamicin: 20,000 units / 500ml;

[0128] 14. Autologous tumor-associated whole antigen: 50ug / ml;

[0129] 15. IFN-γ: 50ug / 500ml;

[0130] 16. CD3 monoclonal antibody: 25ug / 500ml;

[0131] 17. IL-2: 15ug / 500ml;

[0132] 18. 0.9% sodium chloride: 200ml;

[0133] 19. Human albumin: 2.0g;

[0134] 20. Interleukin-2 (IL-2) for injection: 200,000 U;

[0135] Combination ...

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Abstract

The invention relates to an antitumor cell immunotherapy technology, in particular to preparation of a specific tumor killing cell. The preparation method disclosed by the invention comprises the following steps of: 1, sampling a single prokaryotic cell from peripheral blood; 2, separating a DC (Dendritic Cell) from a T cell; 3, maturing the DC and preparing a DC vaccine; 4, preparing a CIK (Cytokine Induced Killer); 5, preparing a CTL (cytotoxic T lymphocyte); and 6, preparing a specific DC-CIK-CTL cell preparation.

Description

technical field [0001] The invention relates to an anti-tumor cell immunotherapy, in particular to the preparation of a specific tumor-killing cell. Background technique [0002] The incidence of tumors is increasing year by year. Among the three traditional modes of surgery, radiotherapy, and chemotherapy, chemotherapy is the only systemic treatment. There are six major defects, namely, the effectiveness of early-stage cells, severe immunosuppression, and ineffectiveness against tumor stem cells, which severely limit clinical applications. "Cellular immunotherapy", as a brand-new new systemic treatment technology, has been applied clinically, and has achieved good results with high efficiency and few side effects. Great treatment mod. [0003] At present, NK, TIL, CTL, or DC-CIK technologies are often used for tumor cell immunotherapy at home and abroad. Although the clinical efficacy is much higher than that of chemotherapy, the reinfused effector cells remain in the bod...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K35/14C12N5/0784C12N5/0783C12N5/078A61P35/00
Inventor 蔡建辉马云龙杰弗里·梅德因
Owner 玥特农生物科技河北有限责任公司
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