阅读说明：本技术 一种细胞免疫疗法评价方法及系统 (Cellular immunotherapy evaluation method and system ) 是由 李家平 谈济洲 于 2020-04-17 设计创作，主要内容包括：本发明涉生物医学技术领域,一种细胞免疫疗法评价方法及系统,所述的一种细胞免疫疗法评价方法包括：获取免疫细胞与肿瘤细胞的图像集合,所述图像集合包括若干帧运动图像；在若干帧运动图像中,将初始帧图像的免疫细胞与肿瘤细胞分别进行特征标记；根据特征标记与若干帧运动图像进行细胞的荧光追踪成像；根据追踪成像结果分析凋亡肿瘤比例。通过本发明所述的一种细胞免疫疗法评价方法及系统可以准确的监测到细胞免疫疗法的疗效。(The invention relates to the technical field of biomedicine, in particular to a cell immunotherapy evaluation method and a cell immunotherapy evaluation system, wherein the cell immunotherapy evaluation method comprises the following steps: acquiring an image set of immune cells and tumor cells, wherein the image set comprises a plurality of frames of moving images; respectively carrying out feature marking on immune cells and tumor cells of an initial frame image in a plurality of frame moving images; performing fluorescence tracking imaging of the cells according to the feature markers and the plurality of frames of moving images; and analyzing the proportion of the apoptotic tumor according to the tracking imaging result. The cellular immunotherapy evaluation method and the cellular immunotherapy evaluation system can accurately monitor the curative effect of the cellular immunotherapy.)

1. A method for evaluating a cellular immunotherapy, comprising:

acquiring an image set of immune cells and tumor cells, wherein the image set comprises a plurality of frames of moving images;

respectively carrying out feature marking on immune cells and tumor cells of an initial frame image in a plurality of frame moving images;

performing fluorescence tracking imaging of the cells according to the feature markers and the plurality of frames of moving images;

and analyzing the proportion of the apoptotic tumor according to the tracking imaging result.

2. The method of claim 1, wherein the immune cells are pre-stained with tumor cells, the immune cells are stained with cell-treated stains, and the tumor cells are stained with caspase3/7 apoptosis probe.

3. The method for evaluating cellular immunotherapy according to claim 1, wherein the method for characterizing immune cells and tumor cells separately comprises:

taking immune cells and tumor cells in the initial frame image as targets, and respectively marking cell nuclei of the two cells as an identifier 1 and an identifier 2;

the shapes of cell membranes of all target cells are sketched, and the cell membranes are marked;

and fusing the images of the cell nucleus and the cell membrane to generate the characteristic labeled cells.

4. The method of claim 3, wherein the step of generating the characteristic labeled cells further comprises color channel decomposition of a color image of the labeled cells:

acquiring an initial frame image, wherein the initial frame image is an RGB image;

and (3) carrying out color channel decomposition on the immune cells and the tumor cells which are stained and marked in the image, and acquiring a color channel image corresponding to each marked cell.

5. The method for evaluating cellular immunotherapy according to claim 1, wherein the performing of cell fluorescence tracking imaging based on the feature labels and the plurality of frames of moving images comprises:

and detecting and identifying each marked cell through a deep learning network, positioning to the position of each target cell, and generating a tracking path.

6. The method for evaluating cellular immunotherapy according to claim 5, wherein the step of detecting and identifying each labeled cell is carried out by,

after each cell finishes the feature marking, simultaneously generating the corresponding RGB value, the cell membrane shape and the cell nucleus shape feature, taking the feature value as a feature point set,

the cells are detected and identified by matching the feature point set,

if the RGB value of the target cell changes, the target cell is not detected, and the subsequent step of detecting the target cell is automatically cancelled.

7. The method for evaluating cellular immunotherapy according to claim 1, wherein the analysis of the proportion of apoptotic tumors according to the three-dimensional imaging result specifically comprises: counting the number of the two marked cells, and judging the number ratio of the two cells.

8. A cellular immunotherapy evaluation system, comprising:

an image acquisition module: the image acquisition module is used for acquiring an image set of immune cells and tumor cells, and the image set comprises a plurality of frames of moving images;

a feature marking module: the characteristic marking module is used for respectively carrying out characteristic marking on immune cells and tumor cells of an initial frame image in a plurality of frame moving images;

a tracking imaging module: the tracking imaging module is used for carrying out fluorescence tracking imaging on the cells according to the feature markers and a plurality of frames of moving images;

an analysis module: the analysis module is used for analyzing the proportion of the apoptotic tumor according to the tracking imaging result.

9. A computer-readable storage medium, characterized in that it stores computer program instructions adapted to be loaded by a processor and to execute the method of any of claims 1 to 7.

10. A mobile terminal comprising a processor and a memory, the processor being configured to execute a program stored in the memory to implement the method of any one of claims 1 to 7.

Technical Field

The invention relates to the technical field of biomedicine, in particular to a cell immunotherapy evaluation method and system.

Background

Cellular immunotherapy of tumors, such as CAR-T, TCR-T, etc., is well-established and increasingly important in tumor therapy. One common feature of both technologies is that the recognition and attack ability of T cell receptors to specific cancer cell antigens is enhanced by means of genetic engineering. And thus are also collectively referred to as "T cell receptor redirection" techniques.

CAR-T and TCR-T are two latest immune cell technologies of the ACT technology for the current adoptive cell-back therapy, and are widely concerned and researched due to the fact that the CAR-T and the TCR-T can express specific receptor target recognition cells such as tumor cells, and the CAR-T and the TCR-T are changed into clinical application from the initial basic immune research. Based on synthetic biology, immunology and genetic modification technology, the synthesis of modified T cells with enhanced specific functions is possible. CD19 antigen-specific CAR-T cells have shown sustained disease remission in clinical trials for the treatment of B cell leukemia and lymphoma. Because of the excellent performance and wide application prospect of the CAR-T/TCR-T technology, the CAR-T/TCR-T technology enters the competitive stage of the current fierce pharmaceutical industry and is higher than the CAR-T/TCR-T technology in the traditional pharmaceutical industry.

However, in the prior art, preclinical efficacy evaluation of cellular immunotherapy has a major drawback, and many patients have poor efficacy in practical clinical application due to lack of preclinical efficacy evaluation. The existing evaluation system is only based on the sequencing result or the pathological result of the whole exon of the patient, and the real killing effect of the immune cells on the tumor is difficult to judge. The personalized PDX model of the patient based on the humanized mouse has extremely high manufacturing cost, great difficulty and long time consumption, and is difficult to be used as a rapid reagent tool for clinical application. In order to solve the above problems, the present invention provides a method and a system for evaluating a cellular immunotherapy.

Disclosure of Invention

The invention solves the technical problem of providing a method and a system for evaluating a cellular immunotherapy. The method and the system for evaluating the cellular immunotherapy can accurately monitor the curative effect of the cellular immunotherapy.

In order to solve the technical problems, the technical scheme provided by the invention is as follows:

a method of evaluating cellular immunotherapy, comprising:

acquiring an image set of immune cells and tumor cells, wherein the image set comprises a plurality of frames of moving images;

respectively carrying out feature marking on immune cells and tumor cells of an initial frame image in a plurality of frame moving images;

performing fluorescence tracking imaging of the cells according to the feature markers and the plurality of frames of moving images;

and analyzing the proportion of the apoptotic tumor according to the tracking imaging result.

Preferably, the immune cells and the tumor cells are labeled by staining in advance, the immune cells are labeled by celltrace-treated staining, and the tumor cells are labeled by staining of caspase3/7 apoptosis probe.

Preferably, the method for separately labeling the immune cells and the tumor cells comprises the following steps:

taking immune cells and tumor cells in the initial frame image as targets, and respectively marking cell nuclei of the two cells as an identifier 1 and an identifier 2;

the shapes of cell membranes of all target cells are sketched, and the cell membranes are marked;

and fusing the images of the cell nucleus and the cell membrane to generate the characteristic labeled cells. The target cells are marked, namely the immune cells are marked as mark 1, and the tumor cells are marked as mark 2, so that each marked cell is ensured to be tracked in the subsequent cell tracking process, and the loss of the cells in the tracking process is prevented.

Further preferably, after the generating of the characteristic labeled cells, the method further comprises performing color channel decomposition on a color image of the labeled cells:

acquiring an initial frame image, wherein the initial frame image is an RGB image;

and (3) carrying out color channel decomposition on the immune cells and the tumor cells which are stained and marked in the image, and acquiring a color channel image corresponding to each marked cell. Because the colors of the two cell markers are different when the cells are marked with the color markers, the RGB values of the two cells are acquired for better marking and distinguishing the two cells, so that the tracking effect is more accurate.

Preferably, the fluorescence tracking imaging of the cell according to the feature marker and the plurality of frames of moving images is specifically:

and detecting and identifying each marked cell through a deep learning network, positioning to the position of each target cell, and generating a tracking path.

Further preferably, the method for detecting and identifying each labeled cell is,

after each cell finishes the feature marking, simultaneously generating the corresponding RGB value, the cell membrane shape and the cell nucleus shape feature, taking the feature value as a feature point set,

the cells are detected and identified by matching the feature point set,

if the RGB value of the target cell changes, the target cell is not detected, and the subsequent step of detecting the target cell is automatically cancelled. In the process of cell tracking, compared data are the shape of cell nucleus, the shape of cell membrane and the specific RGB value of each cell after dyeing, and the cell tracking is carried out on the three aspects, so that the tracking accuracy is ensured.

Preferably, the analyzing the proportion of the apoptotic tumor according to the three-dimensional imaging result specifically comprises: counting the number of the two marked cells, and judging the number ratio of the two cells.

A cellular immunotherapy evaluation system, comprising:

an image acquisition module: the image acquisition module is used for acquiring an image set of immune cells and tumor cells, and the image set comprises a plurality of frames of moving images;

a feature marking module: the characteristic marking module is used for respectively carrying out characteristic marking on immune cells and tumor cells of an initial frame image in a plurality of frame moving images;

a tracking imaging module: the tracking imaging module is used for carrying out fluorescence tracking imaging on the cells according to the feature markers and a plurality of frames of moving images;

an analysis module: the analysis module is used for analyzing the proportion of the apoptotic tumor according to the tracking imaging result.

A computer readable storage medium having stored thereon computer program instructions adapted to be loaded by a processor and to execute a method of cellular immunotherapy evaluation.

A mobile terminal is characterized by comprising a processor and a memory, wherein the processor is used for executing a program stored in the memory so as to realize a cellular immunotherapy evaluation method.

Compared with the prior art, the invention has the beneficial effects that: the cellular immunotherapy evaluation method and the cellular immunotherapy evaluation system can accurately monitor the curative effect of the cellular immunotherapy, are used for preclinical tests of personalized cellular immunotherapy of patients, explore the optimal clinical curative effect to the maximum extent, save the economic cost and the time cost, and have great scientific research and clinical application values. The method specifically comprises the following steps: the method is characterized in that cells are firstly subjected to staining marking and characteristic marking, so that immune cells and tumor cells can be distinguished, the movement paths of the two cells are tracked due to the marking, but when the cells are apoptotic, the cells are not tracked any more due to the staining change, namely the change of RGB values, so that the number of the tumor cells which are not apoptotic can be tracked finally, the apoptotic tumor proportion is determined, each cell can be tracked more accurately through the automatic monitoring of the system, and the optimal clinical curative effect is searched to a great extent.

Drawings

The invention is further illustrated with reference to the following figures and examples.

FIG. 1 is a schematic flow chart of a method for evaluating cellular immunotherapy according to the present invention;

FIG. 2 is a block diagram of an evaluation system of the seed cell immunotherapy according to the present invention.

Detailed Description

The present invention will now be described in further detail with reference to the accompanying drawings. These drawings are simplified schematic drawings and illustrate only the basic flow diagram of the invention, and therefore they show only the flow associated with the invention.