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Method for amplifying and activating CIK lymphocyte

A technology of lymphocytes and cells, applied in the field of immunology, can solve the problems of low killing function of CIK cells, low purity of CD3mAb, and failure to meet clinical needs, etc., to achieve the effect of improving killing ability, improving immunity, and increasing the number

Active Publication Date: 2016-06-22
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The number and killing function of CIK cells are directly related to the curative effect. The problem in the treatment is that it is very difficult to obtain high-purity CIK cells
Traditionally used interleukin 2, IL-1α, IFN-γ, and CD3 mAbs have low purity (20%-30%); the amplified CIK cells have low killing function and cannot meet clinical needs

Method used

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  • Method for amplifying and activating CIK lymphocyte
  • Method for amplifying and activating CIK lymphocyte
  • Method for amplifying and activating CIK lymphocyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Expansion of purified CIK cells.

[0056] Purified CIK cells (1x10 6 , in which CIK cells account for more than 50%) were cultured in RPMI1640 supplemented with 10% fetal bovine serum. Add radiation-killed host cells to the culture medium (after transfecting K562 cells according to the method described in Imai et al. 1α (100IU / ml), IFN-γ (1000IU / ml), CD3mAb (50ng / ml) were cultured for 7 days. Centrifuge after 7 days, resuspend with an equal amount of culture medium, add irradiated host cells, and culture for another 7 days, as figure 1 shown. IL-1α, IFN-γ, CD3mAb and interleukin-2 were used as control groups. Such as figure 1 As shown, the number of CIK cells increased significantly under the joint action of host cells, IL-1α, IFN-γ, CD3mAb and interleukin-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 in cell number were started. Und...

Embodiment 2

[0056] Purified CIK cells (1x10 6 , in which CIK cells account for more than 50%) were cultured in RPMI1640 supplemented with 10% fetal bovine serum. Add radiation-killed host cells to the culture medium (after transfecting K562 cells according to the method described in Imai et al. 1α (100IU / ml), IFN-γ (1000IU / ml), CD3mAb (50ng / ml) were cultured for 7 days. Centrifuge after 7 days, resuspend with an equal amount of culture medium, add irradiated host cells, and culture for another 7 days, as figure 1 shown. IL-1α, IFN-γ, CD3mAb and interleukin-2 were used as control groups. Such as figure 1 As shown, the number of CIK cells increased significantly under the joint action of host cells, IL-1α, IFN-γ, CD3mAb and interleukin-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 in cell number were started. Under the action of host cells:CIK lymphocytes=1:1 conce...

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Abstract

The invention discloses a method for amplifying and activating CIK lymphocyte by using a host cell transfected with functional protein in combination with multiple kinds of protein. The method has the advantages that compared with the prior art, CIK lymphocyte amplified and activated through the method has higher purity, higher amplifying efficiency and higher capability of killing tumor and TAMs in a tumor microenvironment. The method has broad prospects in the aspect of controlling and relieving the progression of part of diseases such as cancer, infectious diseases and diabetes mellitus or the aspects of organ transplantation and hepatitis B.

Description

technical field [0001] The invention belongs to the field of immunology, and specifically refers to a method for directional amplification and activation of CIK lymphocytes by using host cells transfected with various proteins and combining multiple proteins to act together. Background technique [0002] CIK cell therapy has a significant effect on a variety of tumors such as lung cancer, squamous cell carcinoma of the head and neck, sarcoid carcinoma and acute leukemia. At present, CIK cell therapy is mainly used in combination with surgery, chemotherapy and radiotherapy in the United States to solve the problem of tumor recurrence. The number and killing function of CIK cells are directly related to the curative effect. The problem in the treatment is that it is very difficult to obtain high-purity CIK cells. Traditionally used interleukin 2, IL-1α, IFN-γ, and CD3 mAbs have low purity (20%-30%); the amplified CIK cells have low killing function and cannot meet clinical ne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCA61K35/17C12N5/0646C12N2501/2301C12N2501/2302C12N2501/2321C12N2501/24C12N2501/505C12N2501/515C12N2501/599
Inventor 徐以兵郑树梁廷波朱莉莉胡薇蕾
Owner ZHEJIANG UNIV
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