Systemic lupus erythematosus animal model and construction method and application thereof

Abstract

The invention relates to a systemic lupus erythematosus animal model and a construction method and application thereof, and belongs to the technical field of disease animal model construction methods. The animal model construction method for systemic lupus erythematosus comprises the following steps: hybridizing a female mouse of a C57BL / 6 strain as a female parent with a male mouse of an SJL / J strain as a male parent to generate an F1-generation hybrid mouse, and the F1-generation hybrid mouse is more likely to have a lupus phenotype. Compared with a C57BL / 6 female parent, the animal model for systemic lupus erythematosus, provided by the invention, can develop typical symptoms of lupus more quickly and more remarkably, so that the experimental period is greatly shortened, and the research efficiency is improved. Based on the C57BL / 6 background, subsequent fine genetics and mechanism research is facilitated. The method can be obtained through traditional breeding without complex gene operation, is good in stability and high in repeatability, and provides an efficient and reliable new tool for lupus research.

Description

TECHNICAL FIELD

[0001] The present application relates to an animal model of systemic lupus erythematosus and a method for constructing the same and application thereof, and belongs to the technical field of disease animal model construction methods. BACKGROUND

[0002] Current mouse models for systemic lupus erythematosus (SLE) research are mainly divided into four categories: spontaneous models, genetically modified models, induced models, and humanized models.

[0003] Spontaneous models: NZB / W F1 mice: obtained by crossing New Zealand black mice (NZB) and New Zealand white mice (NZW), are the most widely studied model, characterized by the presence of anti-dsDNA and other autoantibodies, and the development of immune complex-mediated glomerulonephritis at 6-8 months of age, with a clear female predisposition. MRL / lpr mice: due to Fas gene mutation leading to lymphocyte proliferation and autoimmunity, with early onset (2-3 months of age), severe phenotype, involving kidney, skin and nervous system, is a commonly used model for studying multiple system involvement in lupus. BXSB mice: characterized by carrying Y chromosome-linked autoimmune accelerator (Yaa), leading to more severe disease in males, associated with upregulation of Toll-like receptor 7 (TLR7) expression. Limitations of classic spontaneous models: classic spontaneous models (such as NZB / W F1, MRL / lpr and BXSB) can mimic some characteristics of human lupus, but their inherent limitations restrict their application in precise mechanism research and drug development: these models generally have complex genetic backgrounds, NZB / W F1 has a long disease period leading to low research efficiency, MRL / lpr disease is driven by a non-typical single Fas gene mutation, and BXSB shows male predominance in disease onset, which is not consistent with the main clinical population. These factors together make it difficult to achieve an ideal balance between genetic controllability and disease phenotype authenticity, and cannot fully meet the growing demand for model tools in modern lupus research.

[0004] Induced models: such as intraperitoneal injection of pristane or skin application of imiquimod The (TLR7 agonist) in normal mice (such as Balb / c, C57BL / 6) to induce a lupus-like phenotype, used to simulate environmental factors triggering.

[0005] Genetically modified models: Constructed by gene editing techniques (such as CRISPR) on C57BL / 6 background, for example, overexpression of TLR7 (Tlr7.Tg) or knockout of FcγRIIB model, for the study of the role of specific genes or pathways. The fundamental defect of C57BL / 6 background model: C57BL / 6 is the "gold standard" for genetic research, but it is inherently low in susceptibility to lupus and difficult to spontaneously form a severe and typical lupus phenotype. Although lupus can be induced on this background through genetic engineering, these models still generally require a longer time to develop significant kidney disease.

[0006] Humanized models: In order to overcome the species differences in immunology between mice and humans, this model reconstructs the human immune system by transplanting human immune cells (such as peripheral blood mononuclear cells (PBMCs)) or hematopoietic stem cells into immunodeficient mice (such as SCID, NSG mice), in order to study human-specific lupus immune responses and pathological processes in mice.

[0007] Limitations and implications of existing hybrid models: Although there are hybrid models such as NZBxSJL F1 (NSF1), they are expensive and most laboratories cannot afford them. Currently, there is no hybrid model that can produce a significant synergistic effect by crossing a strain that is traditionally used for the study of other diseases (such as SJL for experimental autoimmune encephalomyelitis) with a standard strain of C57BL / 6 that is low in susceptibility to lupus, in order to cultivate a new stable strain that combines a clear genetic background and a strong lupus phenotype. SUMMARY

[0008] In view of the deficiencies of the prior art, the purpose of the present application is to provide an animal model of systemic lupus erythematosus and a construction method and application thereof.

[0009] The present application provides an animal model of systemic lupus erythematosus and a construction method thereof, which aims to provide a new mouse model that combines the advantages of a clear and stable C57BL / 6 genetic background and a significantly higher susceptibility to lupus than C57BL / 6, by systematically crossing and breeding C57BL / 6 female mice with SJL / J male mice, in order to overcome the limitations of existing mainstream models in terms of genetic background, modeling period, or phenotype typicality.

[0010] In order to achieve the purpose of the present application, the following technical solution is adopted: A construction method for an animal model of systemic lupus erythematosus, the construction method comprising crossing a C57BL / 6 strain female mouse as a female parent with a male parent to produce F1 hybrid mice, and the F1 hybrid mice spontaneously developing a lupus phenotype.

[0011] In some embodiments of the method for constructing an animal model of systemic lupus erythematosus described in this invention, the male paternal mouse is of the SJL / J strain.

[0012] In some embodiments of the method for constructing an animal model of systemic lupus erythematosus according to the present invention, the specific steps of the method include: (1) Select male SJL / J strain mice as the father and female C57BL / 6J strain mice as the mother to cross and obtain F1 generation hybrid mice. Apply imiquimod cream to the skin of the right ear of each F1 generation hybrid mouse every two days for 4-10 weeks. (2) Detection of urine protein, creatinine and urea nitrogen in F1 generation hybrid mice; Blood was collected from F1 generation hybrid mice to separate plasma, and enzyme-linked immunosorbent assay (ELISA) was performed. After extracting spleen and lymph node tissues from F1 generation hybrid mice, single-cell suspensions were prepared and then subjected to flow cytometry experiments. After extracting kidney tissue from F1 generation hybrid mice, total RNA was extracted, reverse transcribed, and then subjected to real-time quantitative PCR experiments. (3) To establish an animal model of systemic lupus erythematosus and verify whether the F1 generation hybrid mouse systemic lupus erythematosus model has lupus disease susceptibility.

[0013] In some embodiments of the method for constructing an animal model of systemic lupus erythematosus described in this invention, more preferably, step (1) may also be: selecting male SJL / J strain mice as the father and female C57BL / 6J strain mice as the mother to cross them to obtain F1 generation hybrid mice, and injecting phytidine into the F1 generation hybrid mice once via intraperitoneal injection.

[0014] On the other hand, the present invention provides an animal model of systemic lupus erythematosus (SLE), which is constructed using the method for constructing an animal model of SLE described in this invention. Other induction methods may also be used, such as R848 application, allogeneic cell adoptive transfer, or autoantigen inoculation.

[0015] Preferably, the animal model of systemic lupus erythematosus is used in the treatment of systemic lupus erythematosus or in the screening of drugs for the treatment of systemic lupus erythematosus.

[0016] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) Significantly enhanced susceptibility: Compared with the C57BL / 6 parent, the animal model of systemic lupus erythematosus provided by this invention can develop typical lupus symptoms more quickly and significantly, greatly shortening the experimental cycle and improving research efficiency.

[0017] (2) Clear and controllable genetic background: Based on the C57BL / 6 background, the animal model of systemic lupus erythematosus provided by this invention has a clear genetic origin, which facilitates subsequent detailed genetic and mechanistic studies.

[0018] (3) Reliable and efficient modeling: The animal model of systemic lupus erythematosus provided by this invention does not require complex gene manipulation and can be obtained through traditional breeding. It has good stability and strong reproducibility, providing an efficient and reliable new tool for lupus research. Attached Figure Description

[0019] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used in conjunction with embodiments of the invention to explain the invention and do not constitute a limitation thereof. In the drawings: Figure 1 By mating female C57BL / 6 mice with male SJL / J mice, a first-generation hybrid mouse with brown fur was successfully obtained; among them, Figure 1 In this context, A represents a female C57BL / 6 mouse. Figure 1 B in the text refers to male SJL / J mice. Figure 1 C in the text represents generation F1.

[0020] Figure 2 The figure shows the results of using imiquimod to induce a systemic lupus erythematosus model in SJL×C57 F1 mice, compared with C57BL / 6J mice; where, Figure 2 In the figures, A and B represent the upregulation of urinary protein and urinary creatinine levels in SJL×C57 F1 mice. Figure 2 C in the diagram indicates more severe skin lesions on the ear. Figure 2 In this context, D represents the relative increase in ear thickness. Figure 2 E in the text can increase the mRNA levels of inflammatory factors such as TNF-α, IL-1β, and IL-6 in the kidneys; Figure 2 F in the figure represents the results of ELISA detection of antibodies in mouse plasma, which showed that SJL×C57F1 mice had increased levels of anti-dsDNA (IgG), anti-chromatin antibody IgG, and IgG2c. Figure 2 In the figure, G represents immune cells analyzed by flow cytometry. The results showed that the number and proportion of regulatory T cells in the spleen of SJL×C57 F1 mice were decreased. Figure 2 The H in the formula represents an increase in the number of cDC1 cells responsible for antigen cross-presentation, migration to lymph nodes, and activation of T cells. Figure 2 In this context, I represents the number and proportion of plasma cells. Figure 2 The proportion of B220+B cells, plasma cells, and plasmablasts in the J lymph nodes all increased.

[0021] Figure 3The figure shows the results of using phytidine to induce a systemic lupus erythematosus model in SJL×C57 F1 mice, compared with C57BL / 6J mice; among them, Figure 3 In the figure, A and B represent the upregulation of urinary protein and urinary creatinine levels in SJL×C57 F1 mice; Figure 3 C and D in the diagram represent significantly enlarged lymph nodes and spleen. Figure 3 E in the formula can increase the mRNA levels of inflammatory factors such as TNF-α and IL-1β in the kidneys. Figure 3 The F in the figure represents the detection of anti-dsDNA (IgG) in plasma by ELISA, and it was found that SJL×C57 F1 mice can induce higher levels of autoantibodies. Figure 3 The G in the figure represents flow cytometry results showing an increase in the number of follicular cells and Th1 cells associated with SLE pathogenesis in the lymph nodes of SJL×C57 F1 mice. Figure 3 In this context, H represents the cell number of cDC1 and cDC2. Figure 3 The proportion of I cells, B220+B cells, plasmablasts, and plasma cells increased. Detailed Implementation

[0022] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0023] Example 1: Breeding of SJL / J×C57BL / 6 F1 Mice I. Experimental Materials SPF-grade 12-week-old male SJL / J mice (Vitalliwa), 7-week-old female C57BL / 6J mice (Nanjing Huimiaoxin), etc.

[0024] II. Experimental Methods 1. Breeding of SJL×C57 F1 mice 12-week-old male SJL mice were housed together with 7-week-old female C57BL / 6J mice (1 male: 3 females). The vaginal plugs were checked daily or the vaginal opening of the female mice was observed to confirm pregnancy. After pregnancy, the mice were separated into different cages and provided with adequate nutrition and a quiet environment. The gestation period was about 19-21 days. After giving birth, the mother mice were not disturbed. The weaning period for the pups was usually 21-28 days.

[0025] III. Experimental Results By using C57BL / 6 female mice ( Figure 1 .A) and SJL / J males are small ( Figure 1.B) was mated, and its offspring (F1 generation) hybrid mice were successfully obtained. Figure 1 (.C), its fur color is brown.

[0026] Example 2: Imiquimod-induced systemic lupus erythematosus model in SJL / J×C57BL / 6 F1 mice I. Experimental Materials SPF-grade 12-week-old male SJL / J mice (Vitollife), 7-week-old female C57BL / 6J mice (Nanjing Huimiaoxin), imiquimod cream (Sichuan Mingxin Pharmaceutical), urine protein quantification test kit (Nanjing Jiancheng), creatinine (Cr) assay kit (Nanjing Jiancheng), trizol (TaKaRa), reverse transcription reagent (Novizan), sybrgreen (Aikerui), anti-dsDNA (IgG) ELISA kit (Huamei Biotechnology), 4% paraformaldehyde (Biosharp), FOXP3 fixation and permeabilization solution (Invitrogen), flow cytometry antibodies against mouse CD4, CD25, CXCR5, PD-1, Foxp3, Fas, GL-7, MHC-II, CD11c, CD11b, B220, etc. (eBioscience or BioLegend), goat anti-mouse IgG-HRP (Abclonal), goat anti-mouse IgG1-HRP, goat anti-mouse IgG2c-HRP (Southern Biotech), TMB colorimetric solution (Sangon Biotech), etc.

[0027] II. Experimental Methods 1. Imiquimod-induced systemic lupus erythematosus model in SJL×C57 F1 and C57BL / 6J mice Seven-week-old female SJL×C57 F1 and C57BL / 6J mice were treated with 15 mg of 5% imiquimod (imq) cream on the skin of their right ear twice a day for 10 weeks. The mice were then randomly divided into four groups: C57-untreated group (n = 3), C57-imq group (n = 6), SJL×C57 F1-untreated group (n = 3), and SJL×C57 F1-imq group (n = 6).

[0028] 2. Urine protein, creatinine, and blood urea nitrogen tests Test according to the instructions for the Nanjing Jiancheng reagent kits (product numbers: C035-2-1, C011-2-1, C013-2-1).

[0029] 3. Extraction of skin tissue, kidney, spleen, and lymph node tissue Ten weeks later, the mice were euthanized by cervical dislocation after anesthesia. The kidneys, spleen, lymph nodes, and ear skin were removed using ophthalmic scissors and forceps, laid flat on aluminum foil, and flash-frozen in liquid nitrogen or fixed in 4% paraformaldehyde.

[0030] 4. Enzyme-linked immunosorbent assay (ELISA) Plasma was prepared at different time points, aliquoted, and frozen for use in ELISA. A 5 μg / mL chromatin solution was prepared and added to each well at 50 μL. The plate was incubated overnight at 4°C. The next day, the plate was washed three times with washing buffer, and then incubated for 2 hours at room temperature with 200 μL of blocking buffer (1% BSA in PBS). After washing three times, the plate was patted dry. Diluted plasma was added to each well at 50-100 μL and incubated for 2 hours at room temperature. After washing three times and patting dry, goat anti-mouse IgG-HRP, goat anti-mouse IgG1-HRP, or goat anti-mouse IgG2c-HRP (1:5000) was added and incubated for 2 hours at room temperature. After washing 5-7 times and patting dry, TMB chromogenic buffer was added and the plate was developed at room temperature for 30 minutes. The OD value at 450 nm was measured using a microplate reader.

[0031] 5. Enzyme-linked immunosorbent assay (ELISA) for anti-dsDNA (IgG) and other substances. Plasma was prepared at different time points, aliquoted, frozen, and used for ELISA. Its content was determined according to the instructions of the anti-dsDNA (IgG) kit (Huamei Biotechnology, CSB-E11194m).

[0032] 6. Cell extraction and flow cytometry Ten weeks after mouse modeling, spleens and lymph nodes were isolated, ground, filtered to obtain single-cell suspensions, and erythrocytes were removed with erythrocyte lysis buffer. After centrifugation and resuspending, cells were counted. After blocking FCR, antibodies against mouse CD4, CD25, CXCR5, PD-1, Fas, GL-7, MHC-II, CD11c, CD11b, and B220 were added, and the cells were incubated at room temperature in the dark for 20 minutes. Excess antibodies were washed away, and the cells were fixed and nucleated. After nucleation, antibodies against mouse Foxp3, IL-10, and IFN-γ were added, and the cells were incubated at room temperature in the dark for 30 minutes. Data were collected using a Cytek NL Aurora or BD Fortessa flow cytometer and analyzed using FlowJo software.

[0033] 7. Real-time quantitative PCR experiment Total RNA was extracted from kidney and spleen tissues using Trizol reagent (TaKaRa), and RNA quality and concentration were determined using a NanoDrop spectrophotometer (NanoDrop One / OneC, Thermo Fisher Scientific). mRNA was reverse transcribed using a HiScript IV All-in-One Ultra RT SuperMix for qPCR (Vazyme, R433-01), with 1 µg of total RNA used per reaction. qPCR was performed using the SYBR Green SupTaq HS premixed qPCR kit (Accurate Biology) and detected using a LightCycler 96 (Roche). β-actin was used as an internal reference gene, and the mRNA expression levels of the target gene were normalized. The relative expression levels were calculated using the 2^(–ΔΔCt) method.

[0034] III. Experimental Results The inventors' research found that, using imiquimod to induce a systemic lupus erythematosus model in SJL×C57 F1 mice, compared with C57BL / 6J mice, the levels of urinary protein and urinary creatinine in SJL×C57 F1 mice were upregulated. Figure 2 A, B), the skin lesions on the ear are more severe ( Figure 2 .C), relative ear thickness increased ( Figure 2 .D), and can increase the mRNA levels of inflammatory factors such as TNF-α, IL-1β, and IL-6 in the kidneys ( .D), and can increase the mRNA levels of these inflammatory factors ( .D). Figure 2 ELISA detection of antibodies in mouse plasma revealed increased levels of anti-dsDNA (IgG), anti-chromatin antibody IgG, and IgG2c in SJL×C57 F1 mice. Figure 2 Flow cytometry analysis of immune cells showed that the number and proportion of regulatory T cells in the spleen of SJL×C57 F1 mice were decreased. Figure 2 .G), responsible for antigen cross-presentation, migration to lymph nodes, and activation of T cell cDC1 numbers, increases ( Figure 2 .H), the number and proportion of plasma cells ( Figure 2 The proportions of B220+ B cells, plasma cells, and plasmablasts in lymph nodes all increased (I). Figure 3 The above data indicate that the imiquimod-induced SJL×C57 F1 mouse model of systemic lupus erythematosus has a significantly stronger susceptibility to lupus disease than the C57BL / 6 control group.

[0035] Example 3: Phytane-induced systemic lupus erythematosus model in SJL / J×C57BL / 6 F1 mice I. Experimental Materials SPF-grade 12-week-old male SJL / J mice (Vitalliwa), 7-week-old female C57BL / 6J mice (Nanjing Huimiaoxin), sigma (sigma), animal wound suture glue (TIGEROENE), urine protein quantification kit (Nanjing Jiancheng), creatinine (Cr) assay kit (Nanjing Jiancheng), trizol (TaKaRa), reverse transcription reagent (Novizan), sybrgreen (Aikerui), anti-dsDNA (IgG) ELISA kit (Huamei Biotechnology), FOXP3 fixation and permeation solution (Invitrogen), mouse CD4, CD25, CXCR5, PD-1, Foxp3, Fas, GL-7. MHC-II, CD11c, CD11b, B220 and other flow cytometry antibodies (eBioscience or BioLegend), 4% paraformaldehyde (Biosharp), goat anti-mouse IgG-FITC, goat anti-mouse IgG-HRP (Abclonal), goat anti-mouse IgG1-HRP, goat anti-mouse IgG2c-HRP (Southern Biotech), TMB chromogenic solution (Sangon Biotech), etc.

[0036] II. Experimental Methods 1. Phytane-induced systemic lupus erythematosus model in SJL×C57 F1 and C57BL / 6J mice Female SJL×C57 F1 and C57BL / 6J mice were injected intraperitoneally with 0.5 mL of pristane at 7 weeks of age and sacrificed 5 months later. The mice were then randomly divided into 4 groups: C57-untreated group (n = 3), C57-pristane group (n = 5), SJL×C57 F1-untreated group (n = 3), and SJL×C57 F1-pristane group (n = 5).

[0037] 2. Urine protein, creatinine, and blood urea nitrogen tests Test according to the instructions for the Nanjing Jiancheng reagent kits (product numbers: C035-2-1, C011-2-1, C013-2-1).

[0038] 3. Extraction and histological examination of skin tissue, kidney, spleen, and lymph node tissue. Five months later, the mice were euthanized by cervical dislocation after anesthesia. The kidneys, spleen, lymph nodes, and ear skin were removed using ophthalmic scissors and forceps, laid flat on aluminum foil, and flash-frozen in liquid nitrogen or fixed in 4% paraformaldehyde.

[0039] 4. Enzyme-linked immunosorbent assay (ELISA) Plasma was prepared at different time points, aliquoted, and frozen for use in ELISA. A 5 μg / mL chromatin solution was prepared and added to each well at 50 μL. The plate was incubated overnight at 4°C. The next day, the plate was washed three times with washing buffer, and then incubated for 2 hours at room temperature with 200 μL of blocking buffer (1% BSA in PBS). After washing three times, the plate was patted dry. Diluted serum was added to each well at 50-100 μL and incubated for 2 hours at room temperature. After washing three times and patting dry, goat anti-mouse IgG-HRP, goat anti-mouse IgG1-HRP, or goat anti-mouse IgG2c-HRP (1:5000) was added and incubated for 2 hours at room temperature. After washing 5-7 times and patting dry, TMB chromogenic buffer was added and the plate was developed at room temperature for 30 minutes. The OD value at 450 nm was measured using a microplate reader.

[0040] 5. Enzyme-linked immunosorbent assay (ELISA) for anti-dsDNA (IgG) and other substances. Plasma was prepared at different time points, aliquoted, frozen, and used for ELISA. The anti-dsDNA (IgG) content was detected according to the kit instructions.

[0041] 6. Cell extraction and flow cytometry Five months after mouse modeling, spleens and lymph nodes were isolated, ground, filtered to obtain single-cell suspensions, and erythrocytes were removed with erythrocyte lysis buffer. After centrifugation and resuspending, cells were counted. After blocking FCR, antibodies against mouse CD4, CD25, CXCR5, PD-1, Fas, GL-7, MHC-II, CD11c, CD11b, and B220 were added, and the cells were incubated at room temperature in the dark for 20 minutes. Excess antibodies were washed away, and the cells were fixed and nucleated. After nucleation, antibodies against mouse Foxp3, IL-10, and IFN-γ were added, and the cells were incubated at room temperature in the dark for 30 minutes. Data were collected using a Cytek NL Aurora or BD Fortessa flow cytometer and analyzed using FlowJo software.

[0042] 7. Real-time quantitative PCR experiment Total RNA was extracted from kidney and spleen tissues using Trizol reagent (TaKaRa), and RNA quality and concentration were determined using a NanoDrop spectrophotometer (NanoDrop One / OneC, Thermo Fisher Scientific). mRNA was reverse transcribed using a HiScript IV All-in-One Ultra RT SuperMix for qPCR (Vazyme, R433-01), with 1 µg of total RNA used per reaction. qPCR was performed using the SYBR Green SupTaq HS premixed qPCR kit (Accurate Biology) and detected using a LightCycler 96 (Roche). β-actin was used as an internal reference gene, and the mRNA expression level of the target gene was normalized, with its relative expression level calculated using the 2^(–ΔΔCt) method.

[0043] III. Experimental Results Using phytidine to induce a systemic lupus erythematosus model in SJL×C57 F1 mice, compared with C57BL / 6J mice, the levels of urinary protein and urinary creatinine in SJL×C57 F1 mice were upregulated. Figure 3 (A, B) The size of the lymph nodes and spleen is significantly enlarged. Figure 3 (C, D), and can increase the mRNA levels of inflammatory factors such as TNF-α and IL-1β in the kidneys ( Figure 3 E). ELISA detection of anti-dsDNA (IgG) in plasma revealed that SJL×C57 F1 mice could induce higher levels of autoantibodies ( ). Figure 3 Flow cytometry results showed an increase in the number of follicular cells and Th1 cells associated with SLE pathogenesis in the lymph nodes of SJL×C57 F1 mice. Figure 3 .G), the number of cells in cDC1 and cDC2 ( Figure 3 .H) and the proportions of B220+B cells, plasmablasts, and plasma cells ( ​ I) Increased. Experimental data showed that, compared with C57BL / 6 mice, the F1 generation hybrid mice obtained by this protocol exhibited significantly enhanced susceptibility to lupus disease.

[0044] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A method for constructing an animal model of systemic lupus erythematosus, characterized in that, The construction method involves using female C57BL / 6 mice as the maternal parent and crossing them with male mice as the paternal parent to produce F1 generation hybrid mice, which are more prone to lupus phenotype.

2. The method for constructing an animal model of systemic lupus erythematosus as described in claim 1, characterized in that, The male paternal line is the SJL / J strain.

3. The method for constructing an animal model of systemic lupus erythematosus as described in claim 1, characterized in that, The specific steps of the construction method include: (1) Male SJL / J mice were selected as the father and female C57BL / 6J mice as the mother to cross and obtain F1 hybrid mice. Imiquimod cream was applied to the skin of the right ear of each F1 hybrid mouse every two days for 4-10 weeks. (2) Detection of urine protein, creatinine and urea nitrogen in F1 generation hybrid mice; Blood was collected from F1 generation hybrid mice to separate plasma for enzyme-linked immunosorbent assay (ELISA). After extracting spleen and lymph node tissues from F1 generation hybrid mice, single-cell suspensions were prepared for flow cytometry experiments. After extracting kidney tissue from F1 generation hybrid mice, RNA was extracted, reverse transcribed, and then subjected to real-time quantitative PCR experiments. (3) To establish an animal model of systemic lupus erythematosus and verify whether the F1 generation hybrid mouse systemic lupus erythematosus model has lupus disease susceptibility.

4. The method for constructing an animal model of systemic lupus erythematosus as described in claim 3, characterized in that, Step (1) can also be: selecting male SJL / J strain mice as the father and female C57BL / 6J strain mice as the mother to cross them to obtain F1 generation hybrid mice, and injecting phytidine into the F1 generation hybrid mice once via intraperitoneal injection.

5. An animal model of systemic lupus erythematosus, characterized in that, The animal model of systemic lupus erythematosus is constructed by the method for constructing an animal model of systemic lupus erythematosus as described in any one of claims 1-4, or by other induction methods such as R848 application, allogeneic cell adoptive transfer, or autoantigen inoculation.

6. The application of the animal model of systemic lupus erythematosus as described in claim 5 in the treatment of systemic lupus erythematosus or in the screening of drugs for the treatment of systemic lupus erythematosus.

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