WO2020143043A1

WO2020143043A1 - Method for identifying and killing target cell

Info

Publication number: WO2020143043A1
Application number: PCT/CN2019/071418
Authority: WO
Inventors: 周双念
Original assignee: 深圳市双科生物科技有限公司
Priority date: 2019-01-11
Filing date: 2019-01-11
Publication date: 2020-07-16

Abstract

Provided is a method for identifying and killing target cells, comprising: cultivating NK cells having high expression of NKg2d; cultivating CTL cells having high activity; and injecting the NK cells having high expression of NKg2d and the CTL cells having high activity into an abnormal target cell sample. In the microenvironment of the abnormal target cell sample, a part of NKg2d secreted by the NK cells is inserted on the surface of the same kind of CTL cells so as to restore the identification and killing ability of the CTL cells and realize the identification, killing and removal of the target cells positive to MHC-1 type molecules by CTL cells, and at the same time, the non-MHC-1 restrictive characteristic of the NK cells is used for identifying, killing and removing target cells that are negative to non-MHC-1 molecules.

Description

Method for identifying and killing target cells

Technical field

The invention relates to the field of cells, in particular to a method for identifying and killing target cells.

Background technique

In samples with abnormal target cells, the recognition receptor NKg2d on the surface of CTL cells is easily shed, resulting in a decline in the recognition performance of abnormal target cells, and it is difficult to kill abnormal target cells that lowly express MHC-1 molecules.

Summary of the invention

In order to overcome the shortcomings of the prior art, the object of the present invention is to provide a method for identifying and killing target cells to identify and kill abnormal target cells.

The purpose of the present invention is achieved by the following technical solutions:

A method for identifying and killing target cells, including:

Cultivate NK cells with high expression of NKg2d;

Cultivate CTL cells with high activity;

NK cells with high expression of NKg2d and CTL cells with high activity are injected into the abnormal target cell sample, so that part of NKg2d adheres to the CTL cells.

Further, before cultivating NK cells with high expression of NKg2d, the method further includes:

NK cell samples and CTL cell samples were extracted from the samples.

Further, after extracting the NK cell sample and the CTL cell sample from the sample, the method further includes:

The NK cell sample and the CTL cell sample are separated, incubated and expanded.

Compared with the prior art, the beneficial effect of the present invention is that: NK cells with high expression of NKg2d and CTL cells with high activity are injected into abnormal target cell samples, so that part of NKg2d adheres to CTL cells, so that CTL cells release granzyme , Lysosome, perforin, Fas, IFN, etc., to achieve the recognition, killing and clearance of CTL cells to MHC-1 type positive target cells, while applying the non-MHC-1 restrictive characteristics of NK cells, to non-MHC-1 Molecular-negative target cells are identified, killed, and removed.

BRIEF DESCRIPTION

FIG. 1 is a method for identifying and killing target cells according to an embodiment of the present invention.

detailed description

In the following, the present invention will be further described in conjunction with the drawings and specific implementations. It should be noted that, without conflict, the various embodiments described below or the technical features can be arbitrarily combined to form new embodiments. .

As shown in FIG. 1, the method for identifying and killing target cells provided by the embodiments of the present invention includes:

Step S101: Cultivate NK cells with high expression of NKg2d.

Preferably, before culturing NK cells, a NK cell sample and a CTL cell sample are extracted from the sample, wherein the sample and the target cell are from the same subject, the NK cells in the sample normally express NKg2d, and the surface of the CTL cell has NKg2d. After the NK cell sample and the CTL cell sample are extracted, the NK cell sample and the CTL cell sample are separated, cultivated and expanded, and NK cells that highly express NKg2d are cultivated by the existing cultivation method.

Step S102: Cultivate CTL cells with high activity.

Specifically, CTL cells with high activity are cultivated to effectively bind CTL cells to NKg2d.

Step S103: Inject NK cells with high expression of NKg2d and CTL cells with high activity into the abnormal target cell sample, so that part of NKg2d adheres to the CTL cells.

Specifically, abnormal target cells are prone to parasitize or accumulate in solid tissues or organs. The performance of CTL cells in samples of abnormal target cells decreases, and NKg2d on the surface falls off, which cannot kill the mutant cells. NK cells with high expression of NKg2d and CTL cells with high activity are injected into the sample of abnormal target cells successively. In the environment of the sample of abnormal target cells, part of NKg2d secreted by NK cells is adaptively mounted or automatically embedded on the surface of the same kind of CTL cells. In order to improve the ability of CTL cells to recognize abnormal target cells, at the same time release granzyme, lysosome, perforin, Fas, IFN, etc., using the targeting of CTL cells to identify, kill, and kill target cells positive for MHC-1 molecules Clearance: At the same time, use the non-MHC-1 restrictive characteristics of NK cells to identify, kill and clear non-MHC-1 molecule-negative target cells, so as to identify, kill and clear different types and properties of target cells.

The method for identifying and killing target cells provided by the present invention injects NK cells with high expression of NKg2d and CTL cells with high activity into abnormal target cell samples, so that part of NKg2d adheres to CTL cells and restores the recognition and killing ability of CTL cells , So that CTL cells release granzymes, lysosomes, perforin, Fas, IFN, etc., to achieve the recognition, killing and clearance of CTL cells to MHC-1 class positive target cells, while applying the non-MHC-1 restriction of NK cells Sexual characteristics, identification, killing and removal of non-MHC-1 molecule-negative target cells.

The above-mentioned embodiments are only preferred embodiments of the present invention, and cannot be used to limit the scope of protection of the present invention. Any non-substantial changes and replacements made by those skilled in the art on the basis of the present invention belong to the present invention. The scope of protection.

Claims

A method for identifying and killing target cells, characterized in that it includes:

Cultivate NK cells with high expression of NKg2d;

Cultivate CTL cells with high activity;

NK cells with high expression of NKg2d and CTL cells with high activity are injected into abnormal target cell samples, so that part of NKg2d adheres to the surface of the CTL cells.
The method for identifying and killing target cells according to claim 1, wherein before culturing NK cells with high expression of NKg2d, the method further comprises:

NK cell samples and CTL cell samples were extracted from the samples.
The method for identifying and killing target cells according to claim 2, wherein after separating the NK cell sample and the CTL cell sample from the sample, the method further comprises:

The NK cell sample and the CTL cell sample are separated, incubated and expanded.