Method for detecting activity of NK (Natural Killer) cell

Abstract

The invention relates to a method for detecting the activity of an NK (Natural Killer) cell. The method comprises the following steps: specifically expressing a fluorescent protein through a target cell by virtue of a transfection technology to obtain a transfected target cell; co-incubating the NK cell to be detected and the transfected target cell to obtain first mixed cell sap; adding a first fluorescent dye to the first mixed cell sap, and incubating to obtain second mixed cell sap; adding a second fluorescent dye to the second mixed cell sap, and mixing to obtain third mixed cell sap; detecting the third mixed cell sap by a flow cytometer to obtain the cell count ratio of live target cells to early apoptotic target cells to finally apoptotic target cells to dead target cells, wherein the first fluorescent dye is a fluorescein-labeled phospholipid binding protein, the second fluorescent dye is a nucleic acid dye, and the emission wavelengths of the fluorescent protein, the first fluorescent dye and the second fluorescent dye are different from each other. The method for detecting the activity of the NK cell, which is disclosed by the invention, can integrally reflect the activity of the NK cell.

Description

Technical field [0001] The present invention relates to the field of cell detection, in particular to a method for detecting the activity of NK cells. Background technique [0002] Natural killer cell (NK) is an important effector cell in the body's natural immune system. It is not only related to anti-tumor and anti-viral infections, but also has immunoregulatory functions. In some cases, it participates in hypersensitivity and autoimmune diseases. happened. The detection of NK cell activity is an important task in medical research or clinical research. [0003] For the detection of NK cell activity, a variety of detection methods have been developed, such as 51 Cr release method, lactate dehydrogenase (LDH) release method, thiazole blue (MTT) colorimetric method and flow cytometry method, etc. Among them, flow cytometry has attracted more and more attention due to its advantages of no radioactive pollution and high stability. [0004] In related technologies, the commonly used m...

AI Technical Summary

Problems solved by technology

This detection method mainly has the following disadvantages: PI is a nucleic acid fluorescent dye, which can only label late apoptotic cells and dead cells whose cell membranes have lost integrity, so the above detection method cannot reflect the damage caused by NK cells in the process of killing target cells. Proportion of early apoptosis (when cell membrane integrity still exists)

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