A method for detecting NK cell killing activity based on flow cytometry

A technology of flow cytometry and NK cells, which is applied in the field of detection of NK cell killing activity based on flow cytometry, can solve the problems of inability to fully reflect the killing activity of NK cells, and achieve the effect of little interference or influence and a wide range of detection

Active Publication Date: 2021-05-28
XI AN JIAOTONG UNIV
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Problems solved by technology

Although they can specifically detect the apoptosis rate of target cells, they can only mark late apoptotic and necrotic cells, and miss early apoptotic cells, so they cannot fully reflect the killing activity of NK cells

Method used

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  • A method for detecting NK cell killing activity based on flow cytometry
  • A method for detecting NK cell killing activity based on flow cytometry
  • A method for detecting NK cell killing activity based on flow cytometry

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Embodiment Construction

[0037] The present invention is further described below in conjunction with accompanying drawing:

[0038] see Figure 1 to Figure 4 , a method for detecting NK cell killing activity based on flow cytometry, comprising the following steps:

[0039] Step 1, seeding and culturing cells;

[0040] Step 2, collecting cells and staining: using CD2-APC fluorescent dye to group NK cells and tumor cells, and then using Annexin V-FITC / SYTOX Green to stain apoptotic and necrotic target cells;

[0041] Step 3, flow on-board testing.

[0042]Step 1 specifically includes: inoculating and culturing KHYG-1 and K562 cells, changing the liquid for the cells 1-2 days before the experiment, and adjusting the cells to the best state in the logarithmic growth phase; culturing KHYG-1 cells in cells containing IL-2 medium.

[0043] Step 2 includes the following steps:

[0044] 1) Centrifuge NK cells and target cells at 1500rpm for 5 minutes, resuspend and count them separately, and adjust the ce...

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Abstract

A method for detecting NK cell killing activity based on flow cytometry, comprising the following steps: Step 1, seeding and culturing cells; Step 2, collecting cells and staining: using CD2-APC fluorescent dye to group NK cells and tumor cells, Subsequently, Annexin V-FITC / SYTOX Green was used to stain the apoptotic and necrotic target cells; step 3, flow cytometric detection. The present invention uses Annexin V-FITC / SYTOX Green double staining, and uses the same detection channel to mark all the apoptotic cells and necrotic cells, which significantly reduces the various states of tumor cells (early apoptosis, middle and late apoptosis) after killing NK. or necrotic cells).

Description

technical field [0001] The invention belongs to the technical field of cell killing activity detection, in particular to a method for detecting NK cell killing activity based on flow cytometry. Background technique [0002] Natural killer cells (NK cells) are an important part of the body's natural immune system. Under the action of various cytokines (such as IL-2, IL-15, etc.) and feeder cells, NK cells can proliferate and activate, and then release perforin and granzymes to kill tumor cells, viruses and clear non-self cells. NK cells are closely related to tumor immunity, antiviral immunity, transplant immunity and autoimmune diseases. For example, in most tumors (especially those with metastases) and immunodeficiency patients, NK cell activity decreases; while in some viral infectious diseases, long-term use of interferon and its inducers, host resistance to transplantation, etc. In patients with drug reactions, NK cell activity increased. Therefore, accurate evaluati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/14G01N21/64
CPCG01N15/1456G01N21/6428G01N2015/1081G01N2015/1486
Inventor 胡劲松张丹雷莉王爱英
Owner XI AN JIAOTONG UNIV
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