An analysis and evaluation method and system for immunotoxicity

Abstract

The present invention relates to the field of biological immunity, and discloses a method and system for analyzing and evaluating immunotoxicity, the method comprising: culturing immune cells and non-immune cells, constructing an immune microenvironment of immune cell tissue, and constructing an in vitro immune system model of the human body; inputting exogenous substances into the in vitro immune system model, and detecting the cellular immune parameters of the experimental in vitro immune system model; extracting the mouse genome of mice, constructing the gRNA-Cas9 complex of the target immune gene, implanting the gRNA-Cas9 complex into a preset mouse fertilized egg, cultivating the target gene fertilized egg into a target gene mouse, inputting exogenous substances into the body of the target gene mouse, and analyzing the humoral immune parameters of the blood sample; detecting the organ parameters of the immune organ, calculating the organ coefficient of the immune organ, and analyzing the immune organ status of the immune organ; analyzing the immunotoxicity evaluation report of the exogenous substance on the human immune system. The present invention can improve the accuracy of immunotoxicity analysis.

Description

Technical Field

[0001] The present invention relates to the field of biological immunity, and in particular to an analysis and evaluation method and system for immunotoxicity. Background Art

[0002] Immunotoxicity refers to the harmful effects of exogenous substances (such as drugs, chemicals, and environmental pollutants) on the immune system. These effects may manifest as suppression or overactivation of the immune system, leading to a decrease in the body's defenses against pathogens or an excessive immune response. Analysis of immunotoxicity is crucial for drug development, chemical safety assessment, environmental protection, and public health, as it can identify and prevent substances that may have harmful effects on the immune system.

[0003] Currently, immunotoxicity analysis primarily involves administering drugs orally or by injection in experimental animals (such as mice, rats, and rabbits) and then evaluating immune parameters. This approach utilizes animal models, which may not fully mimic the responses of the human immune system, leading to inaccurate predictions. Summary of the Invention

[0004] The present invention provides an immunotoxicity analysis and evaluation method and system, the main purpose of which is to improve the accuracy of immunotoxicity analysis.

[0005] To achieve the above objectives, the present invention provides a method for analyzing and evaluating immunotoxicity, comprising:

[0006] Identifying a human immune system, extracting immune cells and non-immune cells from the human immune system, constructing a cell culture environment for the immune cells and the non-immune cells, culturing the immune cells and the non-immune cells based on the cell culture environment to obtain immune cell tissue, constructing an immune microenvironment for the immune cell tissue, and constructing an in vitro immune system model of the human body based on the immune microenvironment and the immune cell tissue;

[0007] Obtaining exogenous substances, inputting the exogenous substances into the in vitro immune system model to obtain an experimental in vitro immune system model, and detecting cellular immune parameters of the experimental in vitro immune system model;

[0008] obtaining mice, extracting the mouse genome of the mice, determining the target immune gene of the human body, constructing a gRNA-Cas9 complex of the target immune gene based on the mouse genome, implanting the gRNA-Cas9 complex into a preset mouse fertilized egg to obtain a target gene fertilized egg, cultivating the target gene fertilized egg into a target gene mouse, injecting the exogenous substance into the body of the target gene mouse to obtain an experimental mouse, extracting a blood sample from the experimental mouse, and analyzing the humoral immune parameters of the blood sample;

[0009] Identifying the immune organs of the experimental mice, detecting organ parameters of the immune organs, calculating organ coefficients of the immune organs based on the organ parameters, and analyzing the immune organ status of the immune organs based on the organ coefficients;

[0010] Based on the cellular immunity parameters, the humoral immunity parameters and the organ immunity parameters, an immunotoxicity evaluation report of the exogenous substance on the human immune system is analyzed.

[0011] Optionally, constructing a cell culture environment for the immune cells and the non-immune cells comprises:

[0012] constructing a sterile environment for the immune cells and the non-immune cells, and configuring a culture medium for culturing the immune cells and the non-immune cells based on the sterile environment;

[0013] constructing a specific scaffold for the immune cells and the non-immune cells in a culture medium;

[0014] analyzing the gas environments of the immune cells and the non-immune cells, and configuring the oxygen concentration and carbon dioxide concentration of the immune cells according to the gas environments;

[0015] Determining the environmental humidity, environmental temperature, and environmental pH of the immune cells and the non-immune cells;

[0016] Providing the growth factors, hormones and serum required for the development of the immune cells and the non-immune cells;

[0017] A cell culture environment for the immune cells and the non-immune cells is constructed based on the sterile environment, the culture medium, the specific scaffold, the oxygen concentration, the carbon dioxide concentration, the ambient humidity, the ambient temperature, the ambient pH, the growth factors, the hormones and the serum.

[0018] Optionally, constructing the human in vitro immune system model based on the immune microenvironment and immune cell tissue includes:

[0019] Determining the collection time of the immune microenvironment and immune cell tissue;

[0020] Detecting the total number of immune cells in the immune cell tissue and the immune microenvironment at the collection time;

[0021] Detecting the number of infected cells and healthy cells in the immune cell tissue corresponding to the non-immune cells at the collection time;

[0022] determining the cell growth rate of the infected cell population;

[0023] Calculating the elimination rate of the number of infected cells eliminated by the immune cell tissue;

[0024] analyzing the cell death rate of immune cells corresponding to the immune cell tissue;

[0025] Calculating the amount of healthy cells, the amount of infected cells, and the amount of immune cells in the immune cell tissue according to the total immune cell number, the cell growth rate, the elimination rate, and the cell death rate;

[0026] Analyzing the dynamic behavior and cell interactions of the immune cell tissue according to the amount of change in the healthy cells, the amount of change in the infected cells, and the amount of change in the immune cells;

[0027] Based on the dynamic cell behaviors and cell interactions, the human in vitro immune system model is constructed.

[0028] Optionally, the detecting of cellular immune parameters of the experimental in vitro immune system model comprises:

[0029] extracting intracellular signaling molecules from the experimental in vitro immune system model and detecting the molecular activity of the intracellular signaling molecules;

[0030] extracting tissue fluid from the experimental in vitro immune system model, and determining the antibody content and antibody structure of the tissue fluid;

[0031] analyzing immune cell function of the experimental in vitro immune system model based on the antibody content and antibody structure;

[0032] detecting the cell state of the experimental in vitro immune system model and detecting the cytotoxicity of the experimental in vitro immune system model;

[0033] The cellular immune parameters of the experimental in vitro immune system model are determined based on the molecular activity, the immune cell function, the cell state and the cytotoxicity.

[0034] Optionally, extracting the mouse genome of the mouse comprises:

[0035] extracting mouse cells from the mouse, and performing cell lysis on the mouse cells to obtain a mouse cell lysate;

[0036] performing phenol-chloroform extraction on the mouse cell lysate to obtain a mouse genome phenol layer solution;

[0037] Purifying the mouse genome phenol layer solution to obtain a mouse genome precipitation;

[0038] Washing the mouse precipitated genome with a preset alcohol to obtain the mouse genome to be tested;

[0039] The mouse genome quality of the mouse genome to be detected is detected, and when the mouse genome quality meets the preset genome quality standard, the mouse genome to be detected is used as the mouse genome.

[0040] Optionally, constructing the gRNA-Cas9 complex of the target immune gene based on the mouse genome comprises:

[0041] performing genome sequencing on the mouse genome to obtain a mouse genome sequence;

[0042] determining a mouse target sequence of the mouse genomic sequence;

[0043] Performing genome sequencing on the target immune gene to obtain an immune gene sequence;

[0044] According to the mouse target sequence, a gRNA sequence group of the immune gene sequence is constructed, and the gRNA group sequence is transcribed into a gRNA molecule, wherein the gRNA sequence group includes: a guide sequence, a tracrRNA, and a gRNA;

[0045] The preset Cas9 protein and gRNA molecule are combined to obtain the gRNA-Cas9 complex.

[0046] Optionally, the step of cultivating the target gene fertilized egg into a target gene mouse comprises:

[0047] Constructing a culture environment for the target gene fertilized egg;

[0048] Based on the culture environment, the target gene fertilized egg is cultured in vitro to obtain the target gene embryo;

[0049] Implanting the target gene embryo into the uterus of a predetermined adult female mouse to obtain a surrogate mouse;

[0050] When the surrogate mouse completes the pregnancy process, the F1 generation mouse is obtained;

[0051] The F1 generation mouse genes of the F1 generation mice are detected, and target gene mice whose F1 generation mouse genes contain the target immune gene are screened out.

[0052] Optionally, analyzing the humoral immune parameters of the blood sample includes:

[0053] identifying immunoglobulins in the blood sample and determining globulin concentration and subtype distribution of the immunoglobulins;

[0054] analyzing the antibody level of the blood sample based on the globulin concentration and the subtype distribution;

[0055] detecting the cytokine concentration of the blood sample, and analyzing the immune cell status of the experimental mouse corresponding to the blood sample based on the cytokine concentration;

[0056] detecting the level of immunoregulatory molecules in the blood sample, and analyzing the immune system status of the experimental mice based on the level of immunoregulatory molecules;

[0057] The humoral immune parameters of the blood sample are determined based on the antibody level, the immune cell status, and the immune system status.

[0058] Optionally, calculating the organ coefficient of the immune organ based on the organ parameter includes:

[0059] Measuring the body weight of the experimental mice corresponding to the immune organs;

[0060] Extracting immune organ weight from the organ parameters;

[0061] Analyzing the immune weight of the immune organ to the immune system of the experimental mouse;

[0062] The organ coefficient of the immune organ is calculated according to the mouse body weight, the immune organ weight and the immune weight.

[0063] In order to solve the above problems, the present invention also provides an immunotoxicity analysis and evaluation system, which includes:

[0064] An in vitro immune system construction module is used to identify the human immune system of a human body, extract immune cells and non-immune cells of the human immune system, construct a cell culture environment for the immune cells and the non-immune cells, culture the immune cells and the non-immune cells based on the cell culture environment to obtain immune cell tissue, construct an immune microenvironment for the immune cell tissue, and construct an in vitro immune system model of the human body based on the immune microenvironment and the immune cell tissue;

[0065] a cellular immunity analysis module, configured to obtain exogenous substances, input the exogenous substances into the in vitro immune system model to obtain an experimental in vitro immune system model, and detect cellular immunity parameters of the experimental in vitro immune system model;

[0066] A humoral immune analysis module is used to obtain mice, extract the mouse genome of the mice, determine the target immune gene of the human body, construct a gRNA-Cas9 complex of the target immune gene based on the mouse genome, implant the gRNA-Cas9 complex into a preset mouse fertilized egg to obtain a target gene fertilized egg, cultivate the target gene fertilized egg into a target gene mouse, inject the exogenous substance into the body of the target gene mouse to obtain an experimental mouse, extract a blood sample from the experimental mouse, and analyze the humoral immune parameters of the blood sample;

[0067] an immune organ analysis module, configured to identify the immune organs of the experimental mice, detect organ parameters of the immune organs, calculate organ coefficients of the immune organs based on the organ parameters, and analyze the immune organ status of the immune organs based on the organ coefficients;

[0068] The immunotoxicity analysis module is used to analyze the immunotoxicity evaluation report of the exogenous substance on the human immune system based on the cellular immunity parameters, the humoral immunity parameters and the organ immunity parameters.

[0069] The embodiment of the present invention can help study immune cell interactions, immune response mechanisms and immune regulation by constructing the human in vitro immune system model based on the immune microenvironment and immune cell tissue; optionally, the embodiment of the present invention can determine whether the exogenous substance can cause an immune response in the human body by detecting the cellular immune parameters of the experimental in vitro immune system model, which is conducive to analyzing the immune response process in the human body; the embodiment of the present invention can be used to determine the target immune gene of the human body by implanting an experimental animal model for experiment, which is conducive to analyzing the immunotoxicity of the human body; the embodiment of the present invention can analyze the immune response of the experimental mouse to the exogenous substance by analyzing the humoral immune parameters of the blood sample; the embodiment of the present invention can reflect the effect of the exogenous substance on the organ function by calculating the organ coefficient of the immune organ based on the organ parameters. Finally, the embodiment of the present invention can analyze the effect of the exogenous substance on the human immune system by analyzing the immunotoxicity evaluation report of the exogenous substance on the human immune system based on the cellular immune parameters, the humoral immune parameters and the organ immune parameters, thereby accurately judging whether the exogenous substance is immunotoxic. Therefore, the present invention can improve the accuracy of immunotoxicity analysis. BRIEF DESCRIPTION OF THE DRAWINGS

[0070] Figure 1A schematic flow chart of a method for analyzing and evaluating immunotoxicity according to one embodiment of the present invention;

[0071] Figure 2 A functional module diagram of an immunotoxicity analysis and evaluation system provided by one embodiment of the present invention;

[0072] The purpose, features and advantages of the present invention will be further described with reference to the accompanying drawings and in conjunction with the embodiments. DETAILED DESCRIPTION

[0073] It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0074] The embodiment of the present application provides an analysis and evaluation method for immunotoxicity. The execution subject of the analysis and evaluation method of immunotoxicity includes but is not limited to at least one of the electronic devices such as the server, the terminal, etc. that can be configured to execute the method provided by the embodiment of the present application. In other words, the analysis and evaluation method of immunotoxicity can be executed by software or hardware installed in the terminal device or the server device, and the software can be a blockchain platform. The server includes but is not limited to: a single server, a server cluster, a cloud server or a cloud server cluster, etc. The server can be an independent server, or it can be a cloud server that provides cloud services, cloud databases, cloud computing, cloud functions, cloud storage, network services, cloud communications, middleware services, domain name services, security services, content delivery networks (CDN), and basic cloud computing services such as big data and artificial intelligence platforms.

[0075] Reference Figure 1 FIG. 1 is a flow chart of an immunotoxicity analysis and evaluation method according to an embodiment of the present invention. In this embodiment, the immunotoxicity analysis and evaluation method includes:

[0076] S1. Identify the human immune system of the human body, extract immune cells and non-immune cells of the human immune system, construct a cell culture environment for the immune cells and the non-immune cells, culture the immune cells and the non-immune cells based on the cell culture environment to obtain immune cell tissue, construct an immune microenvironment of the immune cell tissue, and construct an in vitro immune system model of the human body based on the immune microenvironment and the immune cell tissue.

[0077] By identifying the human immune system, the present invention can better understand the workings of the immune system and provide a reference for subsequent simulation experiments. The human immune system refers to a biological system responsible for identifying and defending against invading pathogens (such as bacteria, viruses, fungi, and parasites) and abnormal cells (such as cancer cells) in the body.

[0078] Optionally, as an embodiment of the present invention, the human immune system that identifies the human body can be identified through bioinformatics analysis, wherein the bioinformatics analysis refers to a method of analyzing immune-related data using big data and computational methods, such as immune repertoire analysis, network analysis, etc.

[0079] The embodiments of the present invention can be used to simulate the human immune system in vitro and analyze the human immune mechanism by extracting immune cells and non-immune cells of the human immune system. Among them, the immune cells refer to cells that constitute the human immune system and have the ability to recognize and respond to foreign invaders (such as bacteria, viruses, parasites and fungi) and abnormal cells in the body (such as cancer cells), such as B cells, T cells, macrophages, etc. The non-immune cells refer to cells that are not directly involved in the immune response, such as blood cells, nerve cells, fat cells, etc.

[0080] Optionally, as an embodiment of the present invention, the immune cells and non-immune cells of the human immune system can be extracted by magnetic cell sorting, which refers to a technology that uses cell surface specific antigens combined with magnetic particles to separate and purify cells.

[0081] The embodiments of the present invention provide suitable nutrition and environmental conditions by constructing a cell culture environment for the immune cells and the non-immune cells, enabling the immune cells to survive, grow, and proliferate in vitro. The culture environment refers to providing the necessary conditions for the cells to grow, survive, and perform their functions in vitro.

[0082] As an embodiment of the present invention, the step of constructing a cell culture environment for the immune cells and the non-immune cells includes:

[0083] constructing a sterile environment for the immune cells and the non-immune cells, and configuring a culture medium for culturing the immune cells and the non-immune cells based on the sterile environment;

[0084] constructing a specific scaffold for the immune cells and the non-immune cells in a culture medium;

[0085] analyzing the gas environments of the immune cells and the non-immune cells, and configuring the oxygen concentration and carbon dioxide concentration of the immune cells according to the gas environments;

[0086] Determining the environmental humidity, environmental temperature, and environmental pH of the immune cells and the non-immune cells;

[0087] Providing the growth factors, hormones and serum required for the development of the immune cells and the non-immune cells;

[0088] A cell culture environment for the immune cells and the non-immune cells is constructed based on the sterile environment, the culture medium, the specific scaffold, the oxygen concentration, the carbon dioxide concentration, the ambient humidity, the ambient temperature, the ambient pH, the growth factors, the hormones and the serum.

[0089] The environment described herein is one in which there are no living microorganisms (such as bacteria, fungi, viruses, etc.), or the presence of microorganisms is strictly controlled to extremely low levels, insufficient to affect the experimental or production process. The culture medium refers to a medium that provides cells, microorganisms, or other organisms with the necessary nutrients to enable them to grow, reproduce, and / or maintain their biological functions in vitro. The specific scaffold refers to a three-dimensional structure used to support cell growth and differentiation during cell culture. The gas environment refers to the composition and concentration of the gas in which cells are located during cell culture. The environmental humidity refers to the water content in the environment in which cells are located during cell culture. The ambient temperature refers to the ambient temperature conditions in which cells are located during cell culture, generally 37°C (normal human body temperature). The ambient pH refers to the concentration of hydrogen ions (H+) in the environment in which cells are located during cell culture, generally between 7.2 and 7.4. The growth factor is a class of small molecule proteins or polypeptides that can stimulate cell proliferation and differentiation. The hormone refers to a class of chemical substances that promote cell growth and differentiation in cell culture. The serum refers to a liquid that provides cells with rich growth signals and nutrients, used as an additive in many cell culture media.

[0090] Optionally, the specific scaffold for constructing the immune cells and the non-immune cells on the culture medium can be constructed by bioengineering technology, wherein the bioengineering technology refers to the technology of using biological principles and engineering techniques to transform or utilize the structure, function and metabolism of organisms to achieve specific purposes.

[0091] In the embodiments of the present invention, by culturing the immune cells and the non-immune cells in the cell culture environment, the immune cell tissue obtained can simulate the in vivo immune response in vitro and evaluate the effects of exogenous substances on immune cells, including their immunosuppressive or activating effects. The immune cell tissue refers to immune cells aggregated in vitro under in vitro culture conditions through specific culture techniques and methods to form a structure similar to the in vivo immune tissue.

[0092] Optionally, as an embodiment of the present invention, the immune cells and the non-immune cells are cultured based on the cell culture environment to obtain immune cell tissues through bioprinting technology, wherein the bioprinting technology refers to a biomedical application technology that uses computer-aided design (CAD) and computer-aided manufacturing (CAM) technology, combined with biomaterials and cells, to construct a three-dimensional structure by layer-by-layer stacking.

[0093] The embodiments of the present invention can simulate the complexity of the immune system in vivo by constructing the immune microenvironment of the immune cell tissue, providing a model close to the physiological state for studying the interactions between immune cells and immune response mechanisms. The immune microenvironment refers to the environment in vivo or in vitro where immune cells and other cells interact.

[0094] Optionally, as an embodiment of the present invention, the immune microenvironment of the immune cell tissue can be constructed by adding immune regulatory molecules, wherein the immune regulatory molecules include: cytokines, chemokines, etc.

[0095] The embodiments of the present invention construct the human in vitro immune system model based on the immune microenvironment and immune cell tissue, which can help study immune cell interactions, immune response mechanisms, and immune regulation. The in vitro immune system model of the human body is a system model constructed by in vitro culture technology to simulate the structure and function of the human immune system.

[0096] As an embodiment of the present invention, constructing the human in vitro immune system model based on the immune microenvironment and immune cell tissue includes:

[0097] Determining the collection time of the immune microenvironment and immune cell tissue;

[0098] Detecting the total number of immune cells in the immune cell tissue and the immune microenvironment at the collection time;

[0099] Detecting the number of infected cells and healthy cells in the immune cell tissue corresponding to the non-immune cells at the collection time;

[0100] determining the cell growth rate of the infected cell population;

[0101] Calculating the elimination rate of the number of infected cells eliminated by the immune cell tissue;

[0102] analyzing the cell death rate of immune cells corresponding to the immune cell tissue;

[0103] According to the total immune cell number, the cell growth rate, the elimination rate, and the cell death rate, the following formula is used to calculate the healthy cell change, infected cell change, and immune cell change of the immune cell tissue:

[0104]

[0105] in, Indicates the amount of healthy cell changes in immune cell tissue, Indicates the amount of change in immune cell tissue infection cells, represents the change in the number of immune cells in the immune cell tissue, a represents the elimination rate of the number of infected cells by the immune cell tissue, b represents the cell growth rate of the number of infected cells, c represents the cell death rate of the immune cells, K(t) represents the number of healthy cells at the time of collection corresponding to time t, G(t) represents the number of infected cells at the time of collection corresponding to time t, and N(t) represents the total number of immune cells at the time of collection corresponding to time t;

[0106] Analyzing the dynamic behavior and cell interactions of the immune cell tissue according to the amount of change in the healthy cells, the amount of change in the infected cells, and the amount of change in the immune cells;

[0107] Based on the dynamic cell behaviors and cell interactions, the human in vitro immune system model is constructed.

[0108] The collection time refers to the specific time point or time period at which the sample, data, or information is obtained. The total immune cell count refers to the total number of all immune cells in the immune microenvironment and immune tissue at the specific collection time. The infected cell count refers to the total number of cells infected by pathogens in the immune microenvironment and immune tissue at the specific collection time. The healthy cell count refers to the total number of cells not infected by pathogens in the immune microenvironment and immune tissue at the specific collection time. The cell growth rate refers to the rate of increase in cell number over a specific period of time. The elimination rate refers to the rate at which the immune system eliminates infected cells over a specific period of time. The cell death rate refers to the ratio of the number of cell deaths to the initial total number of cells over a specific period of time. The healthy cell change refers to the change in the number of healthy cells in the immune microenvironment and immune tissue over a specific period of time. The infected cell change refers to the change in the number of infected cells in the immune microenvironment and immune tissue over a specific period of time. The immune cell change refers to the change in the number of immune cells in the immune microenvironment and immune tissue over a specific period of time. The dynamic cell behavior refers to the changes in cell behavior under specific environments or conditions. The cell interaction refers to the information transmission and interaction between cells in a variety of ways in an in vivo or in vitro environment.

[0109] Optionally, the detection of the total number of immune cells in the immune cell tissue and the immune microenvironment at the collection time can be performed by an automated cell counter.

[0110] Optionally, the analysis of the dynamic cell behavior and cell interaction of the immune cell tissue according to the change amount of the healthy cells, the change amount of the infected cells and the change amount of the immune cells can be performed by biosensors and nanotechnology.

[0111] S2. Obtain exogenous substances, input the exogenous substances into the in vitro immune system model to obtain an experimental in vitro immune system model, and detect cellular immune parameters of the experimental in vitro immune system model.

[0112] The embodiments of the present invention can activate the immune system by acquiring exogenous substances, inducing immune responses such as immune cell proliferation, differentiation, and cytokine secretion, thereby assessing the function and responsiveness of the immune system. Exogenous substances refer to substances introduced from the outside, which can be harmful pathogens, toxins, vaccines, drugs, or other biologically active substances.

[0113] In the embodiments of the present invention, by introducing the exogenous substance into the in vitro immune system model, an experimental in vitro immune system model is obtained, which can simulate the in vivo infection or immune stimulation process, thereby studying the response of the immune system and the immune response mechanism. Among them, the experimental in vitro immune system model refers to a model for simulating the infection or immune stimulation process of the exogenous substance in the body.

[0114] Optionally, as an embodiment of the present invention, the exogenous substance can be introduced into the in vitro immune system model through a non-viral vector-mediated transfection method, wherein the non-viral vector-mediated transfection refers to the use of non-viral vectors such as liposomes and cationic polymers to introduce exogenous substances into cells.

[0115] The present invention can determine whether a foreign substance can induce an immune response in the human body by detecting cellular immune parameters in the experimental in vitro immune system model, thereby facilitating analysis of the immune response process in the human body. Cellular immune parameters refer to a series of indicators that can quantify and evaluate the function, status, and behavior of immune cells.

[0116] As an embodiment of the present invention, the detecting of cellular immune parameters of the experimental in vitro immune system model includes:

[0117] extracting intracellular signaling molecules from the experimental in vitro immune system model and detecting the molecular activity of the intracellular signaling molecules;

[0118] extracting tissue fluid from the experimental in vitro immune system model, and determining the antibody content and antibody structure of the tissue fluid;

[0119] analyzing immune cell function of the experimental in vitro immune system model based on the antibody content and antibody structure;

[0120] detecting the cell state of the experimental in vitro immune system model and detecting the cytotoxicity of the experimental in vitro immune system model;

[0121] The cellular immune parameters of the experimental in vitro immune system model are determined based on the molecular activity, the immune cell function, the cell state and the cytotoxicity.

[0122] Among them, the intracellular signaling molecules refer to molecules that transmit information inside the cells. For example, protein kinase A, transcription factors, leukotrienes, etc. The molecular activity refers to the ability or efficiency of intracellular signaling molecules in performing their biological functions. The tissue fluid refers to the liquid component used to simulate the in vivo environment in an in vitro immune system model. The antibody content refers to the total amount or concentration of a specific antibody in a given tissue fluid or cell culture supernatant. The antibody structure refers to the molecular structure and composition of the antibody. The immune cell function refers to the various biological functions performed by immune cells in an in vitro model. The cell state refers to the physiological and metabolic state of the cell under a specific environment, which reflects the health, activity and functional state of the cell at that moment. The cytotoxicity refers to the damage or lethality caused by cells or substances produced by cells to other cells.

[0123] Optionally, the detection of the molecular activity of the intracellular signaling molecule can be performed by immunoprecipitation followed by mass spectrometry analysis, wherein the immunoprecipitation followed by mass spectrometry analysis refers to an analysis method that combines immunoprecipitation and mass spectrometry analysis to detect the activity of the signaling molecule.

[0124] Alternatively, the antibody content and antibody structure of the tissue fluid can be determined by enzyme-linked immunosorbent assay (ELISA) to determine the antibody content, and by protein sequencing and X-ray crystallography to determine the antibody structure. The quantitative detection method for determining the antibody content by binding of a specific antibody to an antigen is described. Protein sequencing refers to a method for determining the amino acid sequence of an antibody, and X-ray crystallography refers to a method for analyzing the three-dimensional structure of an antibody.

[0125] Alternatively, the detection of cytotoxicity in the experimental in vitro immune system model can be assessed by a lactate dehydrogenase release assay or a cytotoxicity assay.

[0126] S3. Obtain mice, extract the mouse genome of the mice, determine the target immune gene of the human body, construct the gRNA-Cas9 complex of the target immune gene based on the mouse genome, implant the gRNA-Cas9 complex into a preset mouse fertilized egg to obtain a target gene fertilized egg, cultivate the target gene fertilized egg into a target gene mouse, introduce the exogenous substance into the body of the target gene mouse to obtain an experimental mouse, extract a blood sample from the experimental mouse, and analyze the humoral immune parameters of the blood sample.

[0127] The embodiments of the present invention can be used to establish various experimental models by obtaining mice, providing experimental carriers for subsequent research and analysis.

[0128] The present invention uses genomic data to screen and determine the implantation location of human genes by extracting the mouse genome of the mouse. The mouse genome refers to the collection of all genetic information of the mouse, including all genes, gene regulatory elements, non-coding RNA, repetitive sequences, transposons, and other genetic material of the mouse.

[0129] As an embodiment of the present invention, the step of extracting the mouse genome of the mouse comprises:

[0130] extracting mouse cells from the mouse, and performing cell lysis on the mouse cells to obtain a mouse cell lysate;

[0131] performing phenol-chloroform extraction on the mouse cell lysate to obtain a mouse genome phenol layer solution;

[0132] Purifying the mouse genome phenol layer solution to obtain a mouse genome precipitation;

[0133] Washing the mouse precipitated genome with a preset alcohol to obtain the mouse genome to be tested;

[0134] The mouse genome quality of the mouse genome to be detected is detected, and when the mouse genome quality meets the preset genome quality standard, the mouse genome to be detected is used as the mouse genome.

[0135] Among them, the mouse cell lysate refers to a liquid containing cell lysis products obtained by treating mouse cells by a specific method. The mouse genome phenol layer solution refers to a solution containing the mouse genome obtained by the phenol-chloroform extraction step. The mouse precipitated genome refers to the mouse genome precipitate separated by adding isopropanol for purification to precipitate the mouse genome in the solution. The preset alcohol refers to ethanol with a concentration of 70%. The mouse genome to be tested refers to a sample that needs further testing to determine its quality and purity to meet the needs of the experiment or research. The mouse genome quality refers to a comprehensive evaluation of the characteristics of the mouse genome, such as purity, concentration and integrity. The preset genome quality standard refers to a specific parameter set in the laboratory to evaluate whether the extracted DNA sample is suitable for a specific experiment or research.

[0136] Optionally, the mouse cells may be lysed by using a lysis buffer, such as phenol / chloroform, TRIzol, etc.

[0137] Optionally, purifying the mouse genome phenol layer solution to obtain the mouse genome precipitate comprises:

[0138] performing phenol-chloroform-isoamyl alcohol extraction on the mouse genome phenol layer solution to obtain a mouse genome aqueous phase solution;

[0139] precipitating the mouse genome aqueous solution using a preset isopropanol to obtain a flocculent precipitate mixed solution;

[0140] The flocculent precipitate mixed solution is filtered to obtain the mouse precipitate genome.

[0141] The mouse genome aqueous phase solution contains a mixed solution of the mouse genome. The flocculent precipitation mixed solution refers to a mixed solution of the solid mouse precipitated genome and other impurities.

[0142] The present invention can be used to analyze human immunotoxicity by determining the target immune gene of the human body and implanting it into an experimental animal model. The target immune gene refers to a gene that plays a key role in the function of the human immune system.

[0143] Optionally, the determination of the target immune gene in the human body can be determined by comparative genomics analysis.

[0144] The embodiment of the present invention can implant the target immune gene into the mouse genome by constructing the gRNA-Cas9 complex of the target immune gene based on the mouse genome, so that the target immune gene of the human body is expressed in the mouse body. Wherein, the gRNA-Cas9 complex refers to a complex composed of guide RNA (gRNA) and Cas9 protein.

[0145] As an embodiment of the present invention, the gRNA-Cas9 complex of the target immune gene is constructed based on the mouse genome, comprising:

[0146] performing genome sequencing on the mouse genome to obtain a mouse genome sequence;

[0147] determining a mouse target sequence of the mouse genomic sequence;

[0148] Performing genome sequencing on the target immune gene to obtain an immune gene sequence;

[0149] According to the mouse target sequence, a gRNA sequence group of the immune gene sequence is constructed, and the gRNA group sequence is transcribed into a gRNA molecule, wherein the gRNA sequence group includes: a guide sequence, a tracrRNA, and a gRNA;

[0150] The preset Cas9 protein and gRNA molecule are combined to obtain the gRNA-Cas9 complex.

[0151] Among them, the mouse genome sequence refers to the sequence of all genetic information of the mouse. The mouse target sequence refers to the specific site sequence used to guide the Cas9 protein to perform DNA cutting. The immune gene sequence refers to the DNA sequence of the gene encoding the immune-related protein in the target immune gene. The gRNA sequence refers to the RNA sequence used to guide the Cas9 protein to recognize and cut the mouse target sequence. The guide sequence refers to the 20 bases complementary to the mouse target sequence. The tracrRNA refers to the Cas9 protein, which assists the Cas9 protein in locating the target DNA sequence and cutting. The gRNA molecule refers to the RNA molecule that binds to the Cas9 protein and guides it to the mouse target sequence. The Cas9 protein refers to the protein that binds to the RNA molecule and mediates DNA cutting.

[0152] Optionally, the gRNA sequence group for constructing the immune gene sequence can be constructed by utilizing CRISPR online design tools, such as CRISPR Design, E-CRISPR, etc.

[0153] In the embodiments of the present invention, by implanting the gRNA-Cas9 complex into a pre-determined mouse zygote, a target gene zygote is obtained, which can simulate human immune diseases or study the regulatory mechanism of immune response, thereby helping to reveal the molecular mechanism of immune response. The target gene zygote refers to a zygote in which the mouse zygote gene has been specifically edited.

[0154] Optionally, the implantation of the gRNA-Cas9 complex into a predetermined mouse fertilized egg can be performed by microinjection technology.

[0155] In the embodiments of the present invention, by cultivating the target gene fertilized eggs into target gene mice, the expression and function of the target gene in mice can be studied, which helps to reveal the role of the gene in immunotoxicity. The target gene mouse refers to a mouse model constructed in the laboratory using gene editing technology, in which a specific target gene is knocked out, knocked in, or edited in other ways.

[0156] As an embodiment of the present invention, the step of cultivating the target gene fertilized egg into a target gene mouse comprises:

[0157] Constructing a culture environment for the target gene fertilized egg;

[0158] Based on the culture environment, the target gene fertilized egg is cultured in vitro to obtain the target gene embryo;

[0159] Implanting the target gene embryo into the uterus of a predetermined adult female mouse to obtain a surrogate mouse;

[0160] When the surrogate mouse completes the pregnancy process, the F1 generation mouse is obtained;

[0161] The F1 generation mouse genes of the F1 generation mice are detected, and target gene mice whose F1 generation mouse genes contain the target immune gene are screened out.

[0162] The culture environment refers to an in vitro system that provides suitable growth and development conditions for fertilized eggs. The target gene embryo refers to the early stage of development of the target gene fertilized egg. The surrogate mouse refers to a mouse selected to carry and deliver a gene-edited or modified embryo. The F1 generation mouse refers to the first generation of mice developed from the gene-edited or modified target gene embryo.

[0163] Optionally, the F1 generation mice whose genes contain the target immune gene can be screened out by genetic analysis methods, such as PCR and sequencing.

[0164] In the embodiments of the present invention, by injecting the exogenous substance into the target gene mouse, the obtained experimental mice can be used to study the immune response of the target gene mouse to a specific exogenous substance, which helps to understand the function and regulatory mechanism of the immune system. The experimental mice are mice in which the exogenous substance is injected into the target gene mouse through specific technical means to study specific biological processes.

[0165] Optionally, as an embodiment of the present invention, the introduction of the exogenous substance into the body of the target gene mouse can be performed by injection.

[0166] The embodiment of the present invention can provide analysis samples for subsequent body fluid immune analysis by extracting blood samples from the experimental mice, wherein the blood samples refer to blood samples collected from the experimental mice.

[0167] Optionally, as an embodiment of the present invention, the blood sample of the experimental mouse can be extracted by femoral artery blood sampling, wherein the femoral artery blood sampling refers to a method of collecting blood from the femoral artery of the experimental mouse.

[0168] In the embodiment of the present invention, the immune response of the experimental mice to the exogenous substance can be analyzed by analyzing the humoral immune parameters of the blood sample, wherein the humoral immune parameters refer to the functional status of various biomolecules and immune cells involved in the humoral immune response.

[0169] As an embodiment of the present invention, the analyzing of humoral immune parameters of the blood sample includes:

[0170] identifying immunoglobulins in the blood sample and determining globulin concentration and subtype distribution of the immunoglobulins;

[0171] analyzing the antibody level of the blood sample based on the globulin concentration and the subtype distribution;

[0172] detecting the cytokine concentration of the blood sample, and analyzing the immune cell status of the experimental mouse corresponding to the blood sample based on the cytokine concentration;

[0173] detecting the level of immunoregulatory molecules in the blood sample, and analyzing the immune system status of the experimental mice based on the level of immunoregulatory molecules;

[0174] The humoral immune parameters of the blood sample are determined based on the antibody level, the immune cell status, and the immune system status.

[0175] Among them, the immunoglobulin refers to the antibodies produced in the humoral immune response, such as IgG, IgM, IgA, etc. The subtype distribution refers to the proportion and distribution of different subtypes of immunoglobulins. The antibody level refers to the parameter used to reflect the intensity and type of immune response. The cytokine concentration refers to the content of cytokines in the blood sample. The cytokines are a class of small molecule proteins produced by immune cells and other cells. The immune cell status refers to the function and activity level of immune cells under specific conditions, which reflects the role and status of immune cells in the immune response. The immunoregulatory molecule level refers to the content of immunoregulatory molecules in the blood sample, such as the transforming growth factor-β content, interleukin-10 content, etc. The immune system status refers to the overall function and responsiveness of the immune system under specific conditions.

[0176] Optionally, the determination of the globulin concentration and subtype distribution of the immunoglobulin can be determined by immunoblotting, wherein the immunoblotting refers to a protein analysis technique that separates immunoglobulins into different bands by electrophoresis, then transfers these bands to a solid phase carrier, and then detects the immunoglobulin subtypes on each band by immune reaction.

[0177] Optionally, the cytokine concentration in the blood sample can be detected by real-time quantitative PCR, wherein the real-time quantitative PCR refers to a molecular biology technique that indirectly reflects the cytokine concentration with high sensitivity by designing specific primers and probes to quantitatively analyze the concentration of cytokine mRNA in the sample.

[0178] S4. Identify the immune organs of the experimental mice, detect organ parameters of the immune organs, calculate the organ coefficients of the immune organs based on the organ parameters, and analyze the immune organ status of the immune organs based on the organ coefficients.

[0179] By identifying the immune organs of the experimental mice, the present invention can better understand the structure and function of the immune system and more accurately analyze the stimulating effect of the exogenous substances on the immune system. The immune organs refer to organs in an organism that are specifically involved in immune responses, such as lymph nodes, spleen, thymus, etc.

[0180] The embodiments of the present invention can analyze the physiological state and functional level of immune organs by detecting organ parameters of the immune organs, thereby analyzing the effects of the exogenous substances on the immune organs. The organ parameters refer to a series of indicators used to describe and evaluate the morphology, structure, and function of the organs, such as organ size and organ weight.

[0181] Optionally, the organ parameters of the immune organ can be detected by biometric technology, wherein the biometric technology refers to using a biometric instrument (such as an electronic scale, an ultrasonic measuring instrument, etc.) to measure the size and weight of the immune organ.

[0182] In the embodiment of the present invention, the organ coefficient of the immune organ is calculated based on the organ parameters to reflect the effect of the exogenous substance on the organ function. The organ coefficient refers to the ratio of the weight of the organ to the body weight of the animal.

[0183] As an embodiment of the present invention, the calculating the organ coefficient of the immune organ based on the organ parameter includes:

[0184] Measuring the body weight of the experimental mice corresponding to the immune organs;

[0185] Extracting immune organ weight from the organ parameters;

[0186] Analyzing the immune weight of the immune organ to the immune system of the experimental mouse;

[0187] According to the weight of the mouse, the weight of the immune organ and the immune weight, the organ coefficient of the immune organ was calculated using the following formula:

[0188]

[0189] Where μ represents the organ coefficient, n represents the total number of immune organs, and W i represents the immune organ weight of the i-th immune organ, F i represents the immune weight of the i-th immune organ, W total represents the weight of the mouse, and P represents the proportional coefficient determined according to specific experimental requirements.

[0190] The immune weight refers to the importance of the immune organ in the immune system of the mouse.

[0191] The embodiments of the present invention analyze the immune organ status of the immune organ based on the organ coefficient, which can help analyze the potential impact of the exogenous substance on the immune organ and analyze the functional and structural changes of the immune organ. The immune organ status refers to the function and activity level of the immune organ under specific conditions.

[0192] S5. Analyze the immunotoxicity evaluation report of the exogenous substance on the human immune system based on the cellular immunity parameters, the humoral immunity parameters, and the organ immunity parameters.

[0193] The embodiment of the present invention can analyze the impact of exogenous substances on the human immune system by analyzing the immunotoxicity evaluation report of the exogenous substance on the human immune system based on the cellular immunity parameters, the humoral immunity parameters and the organ immunity parameters, thereby accurately judging whether the exogenous substance is immunotoxic.

[0194] The embodiment of the present invention can help study immune cell interactions, immune response mechanisms and immune regulation by constructing the human in vitro immune system model based on the immune microenvironment and immune cell tissue; optionally, the embodiment of the present invention can determine whether the exogenous substance can cause an immune response in the human body by detecting the cellular immune parameters of the experimental in vitro immune system model, which is conducive to analyzing the immune response process in the human body; the embodiment of the present invention can be used to determine the target immune gene of the human body by implanting an experimental animal model for experiment, which is conducive to analyzing the immunotoxicity of the human body; the embodiment of the present invention can analyze the immune response of the experimental mouse to the exogenous substance by analyzing the humoral immune parameters of the blood sample; the embodiment of the present invention can reflect the effect of the exogenous substance on the organ function by calculating the organ coefficient of the immune organ based on the organ parameters. Finally, the embodiment of the present invention can analyze the effect of the exogenous substance on the human immune system by analyzing the immunotoxicity evaluation report of the exogenous substance on the human immune system based on the cellular immune parameters, the humoral immune parameters and the organ immune parameters, thereby accurately judging whether the exogenous substance is immunotoxic. Therefore, the present invention can improve the accuracy of immunotoxicity analysis.

[0195] like Figure 2 FIG. 1 is a functional module diagram of an immunotoxicity analysis and evaluation system provided by an embodiment of the present invention.

[0196] The immunotoxicity analysis and evaluation system 200 described herein can be installed in an electronic device. Depending on the functionality implemented, the immunotoxicity analysis and evaluation system 200 can include an in vitro immune system construction module 201, a cellular immunity analysis module 202, a humoral immunity analysis module 203, an immune organ analysis module 204, and an immunotoxicity analysis module 205. A module, also referred to as a unit, is a series of computer program segments that can be executed by an electronic device processor and perform a fixed function, and is stored in the electronic device's memory.

[0197] In this embodiment, the functions of each module / unit are as follows:

[0198] The in vitro immune system construction module 201 is used to identify the human immune system of the human body, extract immune cells and non-immune cells of the human immune system, construct a cell culture environment for the immune cells and the non-immune cells, culture the immune cells and the non-immune cells based on the cell culture environment to obtain immune cell tissue, construct an immune microenvironment of the immune cell tissue, and construct the in vitro immune system model of the human body based on the immune microenvironment and the immune cell tissue;

[0199] The cellular immunity analysis module 202 is used to obtain exogenous substances, input the exogenous substances into the in vitro immune system model to obtain an experimental in vitro immune system model, and detect cellular immunity parameters of the experimental in vitro immune system model;

[0200] The humoral immune analysis module 203 is used to obtain mice, extract the mouse genome of the mice, determine the target immune gene of the human body, construct a gRNA-Cas9 complex of the target immune gene based on the mouse genome, implant the gRNA-Cas9 complex into a preset mouse fertilized egg to obtain a target gene fertilized egg, cultivate the target gene fertilized egg into a target gene mouse, inject the exogenous substance into the body of the target gene mouse to obtain an experimental mouse, extract a blood sample from the experimental mouse, and analyze the humoral immune parameters of the blood sample;

[0201] The immune organ analysis module 204 is used to identify the immune organs of the experimental mice, detect organ parameters of the immune organs, calculate the organ coefficients of the immune organs based on the organ parameters, and analyze the immune organ status of the immune organs based on the organ coefficients;

[0202] The immunotoxicity analysis module 205 is used to analyze the immunotoxicity evaluation report of the exogenous substance on the human immune system based on the cellular immunity parameters, the humoral immunity parameters and the organ immunity parameters.

[0203] One embodiment of the present invention provides an electronic device for implementing an analysis and evaluation method for immunotoxicity.

Claims

1. A method for analyzing and evaluating immunotoxicity, characterized in that: The method comprises: Identifying a human immune system, extracting immune cells and non-immune cells from the human immune system, constructing a cell culture environment for the immune cells and the non-immune cells, culturing the immune cells and the non-immune cells based on the cell culture environment to obtain immune cell tissue, constructing an immune microenvironment for the immune cell tissue, and constructing an in vitro immune system model of the human body based on the immune microenvironment and the immune cell tissue; Obtaining exogenous substances, inputting the exogenous substances into the in vitro immune system model to obtain an experimental in vitro immune system model, and detecting cellular immune parameters of the experimental in vitro immune system model; obtaining mice, extracting the mouse genome of the mice, determining the target immune gene of the human body, constructing a gRNA-Cas9 complex of the target immune gene based on the mouse genome, implanting the gRNA-Cas9 complex into a preset mouse fertilized egg to obtain a target gene fertilized egg, cultivating the target gene fertilized egg into a target gene mouse, injecting the exogenous substance into the body of the target gene mouse to obtain an experimental mouse, extracting a blood sample from the experimental mouse, and analyzing the humoral immune parameters of the blood sample; Identifying the immune organs of the experimental mice, detecting organ parameters of the immune organs, calculating organ coefficients of the immune organs based on the organ parameters, and analyzing the immune organ status of the immune organs based on the organ coefficients; Calculating the organ coefficient of the immune organ based on the organ parameter includes: Measuring the body weight of the experimental mice corresponding to the immune organs; Extracting immune organ weight from the organ parameters; Analyzing the immune weight of the immune organ to the immune system of the experimental mouse; Calculating the organ coefficient of the immune organ according to the weight of the mouse, the weight of the immune organ, and the immune weight; According to the weight of the mouse, the weight of the immune organ and the immune weight, the organ coefficient of the immune organ was calculated using the following formula: ； Where μ represents the organ coefficient, n represents the total number of immune organs, and W i represents the immune organ weight of the i-th immune organ, F i represents the immune weight of the i-th immune organ, W total represents the weight of the mouse, and P represents the proportional coefficient determined according to the specific experimental requirements; Analyzing the immunotoxicity evaluation report of the exogenous substance on the human immune system based on the cellular immunity parameters, the humoral immunity parameters, and the organ immunity parameters; constructing the human in vitro immune system model based on the immune microenvironment and immune cell tissue, including: According to the total number of immune cells, cell growth rate, elimination rate and cell death rate, the change in healthy cells, infected cells and immune cells of the immune cell tissue is calculated: ； in, Indicates the amount of healthy cell changes in immune cell tissue, Indicates the amount of changes in immune cell tissue infected cells, represents the amount of change in immune cells in the immune cell tissue, a represents the elimination rate of the infected cells by the immune cell tissue, b represents the cell growth rate of the infected cells, c represents the cell death rate of the immune cells, K(t) represents the number of healthy cells at the acquisition time corresponding to time t, G(t) represents the number of infected cells at the acquisition time corresponding to time t, and N(t) represents the total number of immune cells at the acquisition time corresponding to time t.

2. The immunotoxicity analysis and evaluation method according to claim 1, wherein: The constructing of the cell culture environment for the immune cells and the non-immune cells comprises: constructing a sterile environment for the immune cells and the non-immune cells, and configuring a culture medium for culturing the immune cells and the non-immune cells based on the sterile environment; constructing a specific scaffold for the immune cells and the non-immune cells in a culture medium; analyzing the gas environments of the immune cells and the non-immune cells, and configuring the oxygen concentration and carbon dioxide concentration of the immune cells according to the gas environments; Determining the environmental humidity, environmental temperature, and environmental pH of the immune cells and the non-immune cells; Providing the growth factors, hormones and serum required for the development of the immune cells and the non-immune cells; A cell culture environment for the immune cells and the non-immune cells is constructed based on the sterile environment, the culture medium, the specific scaffold, the oxygen concentration, the carbon dioxide concentration, the ambient humidity, the ambient temperature, the ambient pH, the growth factors, the hormones and the serum.

3. The immunotoxicity analysis and evaluation method according to claim 1, wherein: The method of constructing the human in vitro immune system model based on the immune microenvironment and immune cell tissue comprises: Determining the collection time of the immune microenvironment and immune cell tissue; Detecting the total number of immune cells in the immune cell tissue and the immune microenvironment at the collection time; Detecting the number of infected cells and healthy cells in the immune cell tissue corresponding to the non-immune cells at the collection time; determining the cell growth rate of the infected cell population; Calculating the elimination rate of the number of infected cells eliminated by the immune cell tissue; analyzing the cell death rate of immune cells corresponding to the immune cell tissue; Calculating the amount of healthy cells, the amount of infected cells, and the amount of immune cells in the immune cell tissue according to the total immune cell number, the cell growth rate, the elimination rate, and the cell death rate; Analyzing the dynamic behavior and cell interactions of the immune cell tissue according to the amount of change in the healthy cells, the amount of change in the infected cells, and the amount of change in the immune cells; Based on the dynamic cell behaviors and cell interactions, the human in vitro immune system model is constructed.

4. The immunotoxicity analysis and evaluation method according to claim 1, wherein: The detection of cellular immune parameters of the experimental in vitro immune system model comprises: extracting intracellular signaling molecules from the experimental in vitro immune system model and detecting the molecular activity of the intracellular signaling molecules; extracting tissue fluid from the experimental in vitro immune system model, and determining the antibody content and antibody structure of the tissue fluid; analyzing immune cell function of the experimental in vitro immune system model based on the antibody content and antibody structure; detecting the cell state of the experimental in vitro immune system model and detecting the cytotoxicity of the experimental in vitro immune system model; The cellular immune parameters of the experimental in vitro immune system model are determined based on the molecular activity, the immune cell function, the cell state and the cytotoxicity.

5. The immunotoxicity analysis and evaluation method according to claim 1, wherein: The step of extracting the mouse genome of the mouse comprises: extracting mouse cells from the mouse, and performing cell lysis on the mouse cells to obtain a mouse cell lysate; performing phenol-chloroform extraction on the mouse cell lysate to obtain a mouse genome phenol layer solution; Purifying the mouse genome phenol layer solution to obtain a mouse genome precipitation; Washing the mouse precipitated genome with a preset alcohol to obtain the mouse genome to be tested; The mouse genome quality of the mouse genome to be detected is detected, and when the mouse genome quality meets the preset genome quality standard, the mouse genome to be detected is used as the mouse genome.

6. The immunotoxicity analysis and evaluation method according to claim 1, wherein: The method of constructing the gRNA-Cas9 complex of the target immune gene based on the mouse genome comprises: performing genome sequencing on the mouse genome to obtain a mouse genome sequence; determining a mouse target sequence of the mouse genomic sequence; Performing genome sequencing on the target immune gene to obtain an immune gene sequence; According to the mouse target sequence, a gRNA sequence group of the immune gene sequence is constructed, and the gRNA group sequence is transcribed into a gRNA molecule, wherein the gRNA sequence group includes: a guide sequence, a tracrRNA, and a gRNA; The preset Cas9 protein and gRNA molecule are combined to obtain the gRNA-Cas9 complex.

7. The immunotoxicity analysis and evaluation method according to claim 1, wherein: The method of cultivating the target gene fertilized egg into a target gene mouse comprises: Constructing a culture environment for the target gene fertilized egg; Based on the culture environment, the target gene fertilized egg is cultured in vitro to obtain the target gene embryo; Implanting the target gene embryo into the uterus of a predetermined adult female mouse to obtain a surrogate mouse; When the surrogate mouse completes the pregnancy process, the F1 generation mouse is obtained; The F1 generation mouse genes of the F1 generation mice are detected, and target gene mice whose F1 generation mouse genes contain the target immune gene are screened out.

8. The immunotoxicity analysis and evaluation method according to claim 1, wherein: The analyzing the humoral immune parameters of the blood sample comprises: identifying immunoglobulins in the blood sample and determining globulin concentration and subtype distribution of the immunoglobulins; analyzing the antibody level of the blood sample based on the globulin concentration and the subtype distribution; detecting the cytokine concentration of the blood sample, and analyzing the immune cell status of the experimental mouse corresponding to the blood sample based on the cytokine concentration; detecting the level of immunoregulatory molecules in the blood sample, and analyzing the immune system status of the experimental mice based on the level of immunoregulatory molecules; The humoral immune parameters of the blood sample are determined based on the antibody level, the immune cell status, and the immune system status.

9. An immunotoxicity analysis and evaluation system, characterized in that: A method for analyzing and evaluating immunotoxicity according to any one of claims 1 to 8, wherein the system comprises: An in vitro immune system construction module is used to identify the human immune system of a human body, extract immune cells and non-immune cells of the human immune system, construct a cell culture environment for the immune cells and the non-immune cells, culture the immune cells and the non-immune cells based on the cell culture environment to obtain immune cell tissue, construct an immune microenvironment for the immune cell tissue, and construct an in vitro immune system model of the human body based on the immune microenvironment and the immune cell tissue; a cellular immunity analysis module, configured to obtain exogenous substances, input the exogenous substances into the in vitro immune system model to obtain an experimental in vitro immune system model, and detect cellular immunity parameters of the experimental in vitro immune system model; A humoral immune analysis module is used to obtain mice, extract the mouse genome of the mice, determine the target immune gene of the human body, construct a gRNA-Cas9 complex of the target immune gene based on the mouse genome, implant the gRNA-Cas9 complex into a preset mouse fertilized egg to obtain a target gene fertilized egg, cultivate the target gene fertilized egg into a target gene mouse, inject the exogenous substance into the body of the target gene mouse to obtain an experimental mouse, extract a blood sample from the experimental mouse, and analyze the humoral immune parameters of the blood sample; an immune organ analysis module, configured to identify the immune organs of the experimental mice, detect organ parameters of the immune organs, calculate organ coefficients of the immune organs based on the organ parameters, and analyze the immune organ status of the immune organs based on the organ coefficients; an immunotoxicity analysis module, configured to analyze an immunotoxicity evaluation report of the exogenous substance on the human immune system based on the cellular immunity parameters, the humoral immunity parameters, and the organ immunity parameters; Calculating the organ coefficient of the immune organ based on the organ parameter includes: Measuring the body weight of the experimental mice corresponding to the immune organs; Extracting immune organ weight from the organ parameters; Analyzing the immune weight of the immune organ to the immune system of the experimental mouse; Calculating the organ coefficient of the immune organ according to the weight of the mouse, the weight of the immune organ, and the immune weight; According to the weight of the mouse, the weight of the immune organ and the immune weight, the organ coefficient of the immune organ was calculated using the following formula: ； Where μ represents the organ coefficient, n represents the total number of immune organs, and W i represents the immune organ weight of the i-th immune organ, F i represents the immune weight of the i-th immune organ, W total represents the weight of the mouse, and P represents the proportional coefficient determined according to the specific experimental requirements; Analyzing the immunotoxicity evaluation report of the exogenous substance on the human immune system based on the cellular immunity parameters, the humoral immunity parameters, and the organ immunity parameters; constructing the human in vitro immune system model based on the immune microenvironment and immune cell tissue, including: According to the total number of immune cells, cell growth rate, elimination rate and cell death rate, the change in healthy cells, infected cells and immune cells of the immune cell tissue is calculated: ； in, Indicates the amount of healthy cell changes in immune cell tissue, Indicates the amount of changes in immune cell tissue infected cells, represents the amount of change in immune cells in the immune cell tissue, a represents the elimination rate of the infected cells by the immune cell tissue, b represents the cell growth rate of the infected cells, c represents the cell death rate of the immune cells, K(t) represents the number of healthy cells at the acquisition time corresponding to time t, G(t) represents the number of infected cells at the acquisition time corresponding to time t, and N(t) represents the total number of immune cells at the acquisition time corresponding to time t.

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