Method for amplification and activation of NK cells by K562 cells

A technology of NK cells and cells, applied in the field of immunology, can solve the problems of reduced telomerase activity, low killing function of NK cells, and difficulty in obtaining a large number of NK cells

Active Publication Date: 2013-08-07
杭州中赢生物医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The number and killing function of NK cells are directly related to the curative effect. The problem in the treatment is that it is very difficult to obtain a large number of NK cells
Traditionally used high-dose interleukin-2 activates limited growth folds, reduces telomerase activity, and lowers the killing function of expanded NK cells

Method used

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  • Method for amplification and activation of NK cells by K562 cells
  • Method for amplification and activation of NK cells by K562 cells
  • Method for amplification and activation of NK cells by K562 cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Expansion of NK cells.

[0053] NK cells (2x107) were cultured in RPMI1640 supplemented with 10% human serum. K562 trophoblast cells transfected with CD8α-IL21, CD14, CD19, CD86 and CD137 (irradiated, 100Gy) and IL-2 (50 units / ml) were added to the culture medium and cultured for 7 days. Centrifuge after 7 days, resuspend with an equal amount of culture medium, add irradiated K562 cells transfected with CD8α-interleukin 21, CD14, CD19, CD86 and CD137 (Imai et al. Leukemia, 2004, 18, 676-684), and then Cultured for 7 days, as figure 1 shown. High-dose interleukin-2 (200 units / ml) served as the control group. like figure 1 As shown, the number of NK cells was significantly increased under the combined effect of K562 cells transfected with CD8α-IL-21, CD14, CD19, CD86 and CD137 and low-dose IL-2. An increase in cell number can be measured by the insertion of thymidine. Cells were cultured for 12 hours after thymidine insertion before measurements of changes...

Embodiment 3

[0057] Lysis of tumor cells and TAM (tumor-associated macrophages) by expanded NK cells in Example 3

[0058] Lysis of tumor cells by NK cells expanded under the combined action of K562 trophoblast cells transfected with CD8α-interleukin-21, CD14, CD19, CD86 and CD137 and low-dose interleukin-2

[0059] Previous studies have shown that injecting interleukin-2 into mice with tumors can enhance the anti-tumor ability of mice and delay the lifespan of mice. In this example, K562 trophoblast cells transfected with CD8α-interleukin 21, CD14, CD19, CD86, and CD137 and NK cells expanded under the combined action of low-dose interleukin-2 were co-cultured with tumor cells and tumor-associated macrophages. To study the feasibility of using expanded NK cells for anti-tumor. In this example, K562 trophoblast cells transfected with CD8α-interleukin 21, CD14, CD19, CD86, and CD137 and NK cells expanded under the combined action of low-dose interleukin-2 were injected into mice to treat tu...

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PUM

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Abstract

The invention discloses a method for amplification and activation of NK cells by K562 cells. The method comprises that through synergism of K562 cells transfected by transmembrane interleukin 21, CD14, CD19, CD86 and CD137, and low-concentration interleukin 2, NK cells are subjected to directed amplification and activation. Compared with the existing similar compounds, the compound provided by the invention has a stronger lymphocyte amplification and activation capability and higher efficiency. The method has wide prospects in immunological therapy.

Description

technical field [0001] The invention belongs to the field of immunology, and specifically refers to a method for using K562 cells transfected with transmembrane interleukin 21, CD14, CD19, CD86 and CD137 and low-concentration interleukin 2 to act together to amplify and activate natural killer cells (NK). Background technique [0002] NK cell therapy has a significant effect on the treatment of renal cell carcinoma, melanoma, lung cancer, colon cancer, breast cancer, bladder cancer, liver cancer and acute leukemia. At present, NK cell therapy is mainly used in combination with surgery, chemotherapy and radiotherapy in the United States. The number and killing function of NK cells are directly related to the curative effect. The problem in the treatment is that it is very difficult to obtain a large number of NK cells. Traditionally used high-dose interleukin-2 has limited growth folds, reduced telomerase activity, and reduced killing function of expanded NK cells. The comp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783A61K35/14A61K35/28A61P35/00A61P35/02A61P31/00A61P33/00A61K35/17
CPCY02A50/30
Inventor 吴忠福徐以兵董生聚
Owner 杭州中赢生物医疗科技有限公司
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