Use of anti-igfbp7 antibodies in the treatment of skin diseases

Abstract

The application discloses application of anti-IGFBP7 antibodies in treating skin diseases, belongs to the technical field of psoriasis treatment, and discloses application of anti-IGFBP7 antibodies in preparation of a medicament for treating psoriasis, wherein the medicament is used for reducing and treating psoriasis-like inflammation by inhibiting expression of IGFBP7 in blood vessels of skin of a psoriasis patient. After the anti-IGFBP7 antibodies are used, psoriasis inflammation is reduced, meanwhile, epidermal thickness of a psoriasis patient is reduced, erythema formation is inhibited, and the generation of scales is reduced. In protecting vascular barrier function and treating psoriasis, the anti-IGFBP7 antibodies have a good medicinal prospect.

Description

Technical Field

[0001] This invention relates to the field of psoriasis treatment technology, and more particularly to the application of anti-IGFBP7 antibody in the treatment of skin diseases. Background Technology

[0002] Psoriasis is a common, immune-mediated, chronic inflammatory skin disease. Its stubborn nature places a significant burden on patients, both physically and psychologically, and economically. Psoriasis is characterized by significant vascular abnormalities both clinically and histopathologically. Clinically, scraping away the scales and membranes from the lesions can reveal pinpoint bleeding; histopathologically, the papillary dermal capillaries are tortuous and dilated, surrounded by inflammatory cell infiltration and extravasation of erythrocytes. Although in recent years, biologics or small molecule drugs targeting key cytokines or molecules in the immunopathogenesis of psoriasis have achieved satisfactory therapeutic effects, a complete cure for psoriasis and prevention of recurrence are currently not possible.

[0003] IGFBP7, or insulin-like growth factor binding protein 7, is a secreted glycoprotein with a molecular weight of approximately 30 kDa. Although, as a member of the insulin-like growth factor binding protein family, IGFBP7 has a very low affinity for insulin-like growth factor, only 1 / 1000 to 1 / 100 that of other members, it possesses many functions beyond insulin-like growth factor. Studies have reported that IGFBP7 participates in biological processes such as cellular senescence, cardiac dysfunction, renal failure, and vascular development. It is considered an early biomarker for pulmonary hypertension in patients with systemic sclerosis and, together with TIMP2, serves as a diagnostic biomarker for acute renal failure.

[0004] The expression levels of IGFBP7 in various cell types of normal skin tissue were queried using the public database Human Protein Atlas in single-cell transcriptome sequencing data. In normal skin tissue, IGFBP7 is mainly expressed in smooth muscle cells, fibroblasts, and endothelial cells, while keratinocytes express almost no IGFBP7. This phenomenon was also confirmed by tissue immunofluorescence, indicating that IGFBP7 expression in the epidermis is very low, and there is no significant difference in expression levels between normal human epidermis and psoriasis lesion epidermis. Therefore, it is urgent to investigate the pathogenesis of IGFBP7 in psoriasis treatment and to develop a novel treatment method for psoriasis; finding new therapeutic targets and strategies for psoriasis is particularly necessary. Summary of the Invention

[0005] Therefore, the purpose of this invention is to provide the application of anti-IGFBP7 antibodies in the treatment of skin diseases, which can alleviate and treat psoriasis-like inflammation by inhibiting the expression of IGFBP7 in the blood vessels of the skin of psoriasis patients.

[0006] The present invention solves the above-mentioned technical problems through the following technical means:

[0007] Use of anti-IGFBP7 antibody in the preparation of agents for the treatment of inflammatory skin diseases.

[0008] Furthermore, the use of anti-IGFBP7 antibodies in the preparation of agents for the treatment of psoriasis.

[0009] Furthermore, the use of anti-IGFBP7 antibodies in the preparation of drugs for the treatment of lupus erythematosus.

[0010] Furthermore, the anti-IGFBP7 antibody is an anti-IGFBP7 monoclonal antibody or an anti-IGFBP7 neutralizing antibody.

[0011] Furthermore, the aforementioned treatment for psoriasis is particularly used for treating psoriasis vulgaris.

[0012] Furthermore, the anti-IGFBP7 antibody is mixed with pharmaceutically acceptable excipients to form an injection for the treatment of psoriasis.

[0013] Furthermore, the agent is used to reduce and treat psoriasis-like inflammation.

[0014] Furthermore, the agent is used to alleviate and treat psoriasis-like inflammation by inhibiting the expression of IGFBP7 in the blood vessels of the skin of psoriasis patients.

[0015] Furthermore, when treating psoriasis, the dosage of anti-IGFBP7 antibody is ≥2.5 μg / kg, preferably ≥5 μg / kg.

[0016] Beneficial effects:

[0017] This invention discloses the use of anti-IGFBP7 antibodies in the preparation of medications for inflammatory skin diseases, particularly for treating psoriasis or lupus erythematosus. The anti-IGFBP7 antibody reduces and treats psoriasis-like inflammation by inhibiting IGFBP7 expression in the blood vessels of psoriasis patients. After using the anti-IGFBP7 antibody, the expression of related inflammatory factors in psoriasis patients decreases, psoriasis inflammation is alleviated, and the epidermal thickness of psoriasis lesions is reduced, erythema formation is inhibited, and scaling is reduced. It shows promising pharmaceutical potential in protecting vascular barrier function and treating psoriasis. Attached Figure Description

[0018] Figure 1 : Control diagram of IGFBP7 expression in vascular endothelial cells of skin in psoriasis patients and healthy controls;

[0019] Figure 2 A comparative diagram of IGFBP7 levels in peripheral blood of psoriasis patients and healthy controls;

[0020] Figure 3 : Effect of anti-IGFBP7 antibody on psoriasis-like phenotype in mice;

[0021] Figure 4 Figure: Effect of anti-IGFBP7 antibody treatment on the expression of psoriasis-related inflammatory factors in mouse skin;

[0022] Figure 5 Figure: Effect of anti-IGFBP7 antibody treatment on the expression of vascular barrier molecules in human skin microvascular endothelial cell lines;

[0023] Figure 6 Figure: Effect of anti-IGFBP7 antibody treatment on the adhesion function of human skin microvascular endothelial cell line. Detailed Implementation

[0024] The present invention will be described in detail below with reference to specific embodiments:

[0025] Example 1: Detection of IGFBP7 expression in vascular endothelial cells of skin of psoriasis patients and healthy controls

[0026] (1) Acquisition of human skin tissue and blood samples:

[0027] Skin or blood tissue samples were collected from 46 psoriasis patients and 51 healthy subjects for single-cell transcriptome sequencing, histological and immunofluorescence detection, ultrastructural observation, and molecular expression level detection. The collection of clinical samples and the in vitro experimental research plan were approved by the Ethics Committee of Xijing Hospital of Air Force Medical University (KY20183019-1), and informed consent was obtained from the clinical tissue donors.

[0028] Patients included in this study had psoriasis vulgaris, no other autoimmune or systemic diseases, had never received biologic therapy, and had not undergone any systemic conventional treatment within one month prior to skin or blood tissue sample collection. Skin samples were taken from the site and within a 5cm radius around the collection point. 2 The affected area had not been treated with topical medications for two weeks. Skin and blood tissue samples from psoriasis patients were obtained through routine pathological biopsies and blood tests performed at Xijing Skin Hospital.

[0029] Healthy participants and psoriasis patients were matched for sex and age in this study. Skin tissue samples were obtained from routine plastic surgeries performed at the Department of Plastic Surgery, Department of Urology, and Surgical Center of Xijing Hospital, and were independently assessed by two dermatologists to be free of obvious lesions. Blood samples from healthy participants were obtained through physical examinations or donations.

[0030] (2) Imiquimod-induced mouse psoriasis-like model and sample acquisition:

[0031] Approximately 12 female BALB / c inbred mice were purchased from the Animal Experiment Center of Air Force Medical University and housed in a temperature- and humidity-controlled environment in the SPF-grade laboratory of the Air Force Medical University Animal Experiment Center until they reached 6-8 weeks of age before starting animal experiments. The mice were numbered and randomly divided into a psoriasis group and a control group (N=6 mice / group). Two days before the start of the experiment, the backs of the mice were shaved using an electric shaver, scissors, and depilatory cream (Veet), creating an exposed area of ​​approximately 2×3 cm. This animal experiment was approved by the Experimental Animal Ethics Committee of Air Force Medical University.

[0032] 5% imiquimod cream (50 mg / cm²) was applied topically to the dorsal skin of mice in the psoriasis group. 2 The treatment was administered daily (INova Pharmaceuticals) for 5 days. On day 6, pale red patches and scaling were observed on the mouse skin, indicating successful establishment of the psoriasis mouse model. The control group mice received an equal amount of the cream matrix applied topically to the skin on their backs and the ventral and dorsal sides of their ears. Skin tissue from the backs of both groups of mice was collected on day 6.

[0033] (3) Immunofluorescence assay was used to detect the expression of IGFBP7 in the skin of psoriasis patients and healthy individuals:

[0034] Collected human and mouse skin tissues were fixed with 4% paraformaldehyde (Solepro, cat.no. P1110) and then embedded in paraffin. Sections were prepared using a paraffin microtome and staining machine, with a section thickness of approximately 4 μm. The paraffin sections were automatically dewaxed by immersing them in xylene, followed by immersion in anhydrous ethanol twice, 95% ethanol once, 90% ethanol once, and 75% ethanol once, respectively. Each immersion was then washed twice with distilled water, with each immersion lasting 3-5 minutes.

[0035] The slides were placed in sodium citrate antigen retrieval solution (1×) and incubated at high temperature (98℃) for 20 min, then allowed to return to room temperature. The slides were then washed with PBST for 5 min, repeated three times, and blocked with goat serum using the drop method. After removing excess goat serum, anti-IGFBP7 antibody (1:1000 v / v, Abcam, cat.no.ab74169) and anti-CD31 antibody (1:1000 v / v, Abcam, cat.no.ab199012) were diluted with antibody dilution buffer (NewSemi Biotechnology Co., Ltd., cat.no.WB500D) and incubated overnight (4℃) using the drop method. After the slides were brought to room temperature, they were washed again with PBST for 5 min, and then washed three times. The fluorescently labeled secondary antibody (1:200 v / v, Abcam, cat.no.ab150077, ab97035) was diluted with antibody diluent (NewSemi Biotechnology Co., Ltd., cat.no.WB500D) and incubated for 1 h (room temperature, protected from light) by spotting. The slides were then washed again with PBST for 5 min, and then washed three times. Hoechst 3342 solution (1:1000 v / v, Thermo Fisher Scientific, cat.no.H3570) was diluted with antibody diluent (NewSemi Biotechnology Co., Ltd., cat.no.WB500D) and incubated for 10 min (room temperature, protected from light) by spotting. The slides were then washed again with PBST for 5 min, and then washed three times. Excess liquid was wiped off with filter paper, and the slides were mounted with glycerol and stored at 4°C protected from light. Fluorescence signals from skin tissue were acquired using confocal fluorescence microscopy. The fluorescence intensity of different fluorescence channels was analyzed using ImageJ and Imaris software (version 7.4.2). The results are as follows: Figure 1 As shown.

[0036] (4) ELISA method for detecting the level of IGFBP7 in peripheral blood of psoriasis patients and healthy individuals:

[0037] Peripheral blood was collected from psoriasis patients and healthy subjects using procoagulant blood collection tubes. After standing, the blood was centrifuged (3000 rpm, 5 min), and the supernatant pale yellow serum was collected. Protein standards were prepared according to the ELISA kit (Elabscience, cat.no. E-EL-H0447c) instructions, and the serum samples were diluted. Two to three replicates were prepared for each standard and sample.

[0038] Standards and samples were added to the 96-well plate of the ELISA kit using a multichannel pipette. The standards and samples were incubated according to the conditions recommended in the kit instructions. Primary antibody, HRP, substrate working solution, and stop solution were added sequentially to complete the ELISA procedure. The absorbance of each well in the 96-well plate was measured using a fully automated ELISA analyzer (Bio-Rad). The concentration of IGFBP7 in each sample was calculated based on a linear curve fitted to the standard concentrations. The results are shown below. Figure 2 As shown.

[0039] analyze Figure 1 It can be known that:

[0040] IGFBP7 and CD31 were labeled in skin lesions of psoriasis patients and healthy controls using tissue immunofluorescence (A), with CD31 labeling dermal vascular endothelial cells. The immunofluorescence intensity of IGFBP7 in CD31-positive areas of psoriasis and healthy skin was statistically analyzed. Results showed that IGFBP7 expression in dermal vascular endothelial cells of psoriasis patients was significantly higher than in healthy controls (P<0.0001) (B), while there was no significant difference in IGFBP7 expression in epidermal cells of psoriasis patients compared to healthy controls (ns) (C).

[0041] analyze Figure 2 It can be known that:

[0042] The levels of IGFBP7 in the peripheral blood of 20 psoriasis patients and healthy controls were detected by ELISA. The results showed that the level of IGFBP7 in the peripheral blood of psoriasis patients was significantly higher than that in healthy controls (P = 0.0139) (A), and the concentration of IGFBP7 in the peripheral blood of psoriasis patients was significantly positively correlated with the disease severity score (PASI) of psoriasis patients (P < 0.0001) (B).

[0043] It is evident that IGFBP7 is abnormally highly expressed in the vascular endothelial cells of skin lesions in psoriasis patients, mediating abnormal skin vascularity and skin immune inflammation disorders in psoriasis. This invention aims to alleviate and treat psoriasis-like inflammation by inhibiting the expression of IGFBP7 in the skin blood vessels of psoriasis patients.

[0044] Example 2: Anti-IGFBP7 antibody therapy

[0045] Treatment of psoriasis-like mice with imiquimod-induced anti-IGFBP7 antibody

[0046] (1) Mouse grouping:

[0047] Twenty-four female BALB / c inbred mice were purchased from the Animal Experiment Center of Air Force Medical University and housed in a temperature- and humidity-controlled environment in the SPF-grade laboratory of the Air Force Medical University Animal Experiment Center until they reached 6-8 weeks of age before starting animal experiments. The mice were numbered and randomly divided into four groups: anti-IGFBP7 antibody 2.5 μg / kg group, anti-IGFBP7 antibody 5 μg / kg group, anti-IGFBP7 antibody 7.5 μg / kg group, and IgG control group (N = 6 mice / group). Two days before the experiment, the backs of the mice were shaved using an electric razor, scissors, and depilatory cream to create an exposed area of ​​approximately 2 × 3 cm. This animal experiment was approved by the Experimental Animal Ethics Committee of Air Force Medical University.

[0048] (2) Establishment of a mouse psoriasis-like model and treatment intervention:

[0049] 5% imiquimod cream (50 mg / cm²) was applied topically to the dorsal skin of four groups of mice. 2 The mice were treated with the drug 1 / day (INova Pharmaceuticals) for 5 consecutive days. On day 6 of modeling, pale red patches and scaling were observed on the skin of the mice, indicating successful establishment of the psoriasis mouse model.

[0050] Anti-IGFBP7 antibody was used to treat psoriatic mice. The psoriatic mice were divided into three groups with different doses of treatment: (1) anti-IGFBP7 antibody 2.5 μg / kg group, (2) anti-IGFBP7 antibody 5 μg / kg group and (3) anti-IGFBP7 antibody 7.5 μg / kg group; the IgG control group mice used normal rabbit control IgG antibody (Sinochem Shenzhou Co., Ltd., cat.no.CR1).

[0051] Among them, mice in the anti-IGFBP7 antibody 2.5 μg / kg group, anti-IGFBP7 antibody 5 μg / kg group and anti-IGFBP7 antibody 7.5 μg / kg group were treated with rabbit anti-IGFBP7 monoclonal antibody (ABclonal, cat.no.A4615) to neutralize the target molecule in imiquimod-induced psoriasis-like mice.

[0052] Antibodies administered to all four groups of mice were brought to a final volume of 100 μL with physiological saline and injected intraperitoneally starting one day before the establishment of the psoriasis-like mouse model, once every other day (for a total of 3 times). On day 7 of treatment (i.e., day 6 of imiquimod-induced psoriasis-like mouse model establishment), gross lesions on the back and ears of the mice were photographed using a camera and dermoscopy, and skin tissue from the back of the mice was collected. The results are as follows: Figure 3 As shown.

[0053] (3) Assessment of the severity of psoriasis in mice:

[0054] Mouse dorsal skin was collected, fixed in 4% paraformaldehyde (Solepro, cat.no. P1110), embedded in paraffin, and sectioned. The mouse dorsal skin tissue sections were stained with hematoxylin and eosin. The sections were scanned using a pathological section scanning imaging system, and epidermal thickness was measured using ImageJ software. Ki67 expression in mouse dorsal skin was detected by tissue immunofluorescence assay (method as described above) using anti-Ki67 antibody (Abcam, cat.no. ab15580), and the proportion of Ki67-positive cells in the basal layer of the skin epidermis was analyzed using ImageJ. Total RNA was extracted from mouse dorsal skin specimens using TRIzol reagent (Invitrogen, cat.no. 15596026), and cDNA was obtained by reverse transcription using a reverse transcription kit (Takara, cat.no. RR047A). The expression of S100a8, S100a9, Lcn2, Il17a, Il23, Il36g, Tnfa, and Vegfa was detected by qPCR using a quantitative real-time PCR kit (Takara, cat.no. RR820A). Actb levels were used as a baseline. -ΔΔCT The method was used to calculate relative mRNA expression levels. The results are as follows: Figure 4 As shown.

[0055] analyze Figure 3 It can be known that:

[0056] 1. Compared with the control group treated with IgG, imiquimod-induced psoriasis-like mice treated with anti-IGFBP7 monoclonal antibody showed reduced epidermal thickness. As can be seen from the images, the control group mice had significantly more scales and more obvious erythema symptoms and increased infiltration. This indicates that anti-IGFBP7 antibody treatment can significantly reduce the epidermal thickness of psoriasis, inhibit erythema formation, and reduce scale production.

[0057] 2. Imiquimod-induced psoriasis-like mice were treated with three different doses of anti-IGFBP7 monoclonal antibody (2.5 μg / kg, 5 μg / kg, and 7.5 μg / kg), with a control IgG antibody serving as the control group. Results showed that while the 2.5 μg / kg concentration also alleviated symptoms, the 5 μg / kg and 7.5 μg / kg concentrations significantly increased the therapeutic effect, effectively reducing scaling and erythema (A) in the psoriasis-like mice.

[0058] 3. Treatment with anti-IGFBP7 monoclonal antibodies at doses of 5 μg / kg and 7.5 μg / kg significantly reduced epidermal thickness and the proportion of Ki67 cells in the epidermis of psoriasis-like mice, further demonstrating that anti-IGFBP7 monoclonal antibodies can alleviate psoriasis symptoms and treat psoriasis. Furthermore, the improvement in psoriasis-like mouse phenotype was not significantly different between the 7.5 μg / kg dose and the 5 μg / kg dose (B, C).

[0059] analyze Figure 4 It can be known that:

[0060] 1. It is known that S100a8, S100a9, Lcn2, Il17a, Il23, Il36g, Tnfa, Vegfa, and other inflammatory factors associated with psoriasis are highly expressed in psoriasis patients. Treatment with anti-IGFBP7 monoclonal antibody can effectively reduce the expression of the above-mentioned psoriasis-related inflammatory factors in mouse skin, indicating that anti-IGFBP7 antibody treatment can prevent the aggravation of inflammation in psoriasis patients and thus alleviate psoriasis symptoms.

[0061] 2. In the 5 μg / kg and 7.5 μg / kg dose groups, the reduction of the aforementioned inflammatory factors was most significant, indicating that both 5 μg / kg and 7.5 μg / kg doses are effective in treating psoriasis vulgaris. There was no significant difference in the effect of the 5 μg / kg and 7.5 μg / kg anti-IGFBP7 monoclonal antibody dose groups on the expression levels of psoriasis-related inflammatory factors.

[0062] Therefore, it can be seen that anti-IGFBP7 antibody can significantly reduce psoriasis-like inflammation in mice, protect vascular barrier function and reduce the incidence of psoriasis, and has good medicinal prospects in the treatment of psoriasis.

[0063] Example 3: Human Cell Experiment

[0064] Intervention with anti-IGFBP7 antibody in human skin microvascular endothelial cell line (HMEC-1 cells)

[0065] (1) In vitro culture of HMEC-1 cells:

[0066] HMEC-1 cells (CRL-3243) were purchased from ATCC. HMEC-1 cells were cultured in MCDB131 medium at 37°C and 5% CO2. When the cell density reached 70%–80%, HMEC-1 cells were digested at room temperature using pre-warmed 0.25% trypsin-EDTA digestion buffer (37°C). Cell morphology was observed under a microscope. When 70% of the cells became rounded, digestion was stopped with twice the volume of fetal bovine serum. After centrifugation at room temperature for 5 min (800 rpm), the cell pellet was collected, resuspended in HMEC-1 cell culture medium, and passaged at a ratio of 1:2 to 1:4.

[0067] (2) Intervention with HMEC-1 cells using anti-IGFBP7 antibody:

[0068] HMEC-1 cells were stimulated with recombinant human protein IFN-γ (100 ng / mL, Sigma, cat.no. SRP3058), a psoriasis-associated cytokine, and cell supernatant was collected after 48 h. The newly formed HMEC-1 cells were divided into three groups: Group 1 (control group) received cell supernatant from normal HMEC-1 cells and normal mouse control IgG antibody (25 μg / mL, Abclonal, cat.no. AC011); Group 2 (stimulation group) received cell supernatant from HMEC-1 cells stimulated with psoriasis-associated cytokines and normal mouse control IgG antibody (25 μg / mL, Abclonal, cat.no. AC011); and Group 3 (intervention group) received cell supernatant from HMEC-1 cells stimulated with psoriasis-associated cytokines and mouse anti-IGFBP7 neutralizing antibody (25 μg / mL, Sinocare, cat.no. 13100-MM01).

[0069] (3) HMEC-1 cell barrier structure detection:

[0070] Three groups of HMEC-1 cells were fixed with 4% paraformaldehyde (Solepro, cat.no. P1110) (room temperature, 15 min). After washing three times with PBS, the cells were blocked with goat serum (Boster Biologics, cat.no. AR0009) by drip incubation for 1 h (room temperature). After removing excess goat serum, the anti-hyaluronic acid antibody (1:400 v / v, Abcam, cat.no. ab53842) and the anti-heparan sulfate antibody (1:200 v / v, Abcam, cat.no. ab2501) were diluted with antibody diluent (Synsemi Biotech, cat.no. WB500D) and incubated overnight (4°C) by drip incubation. The cells were brought to room temperature, washed three times with PBS, and the fluorescent secondary antibody (1:200 v / v) was diluted with antibody diluent (Synsemi Biotech, cat.no. WB500D) and incubated for 1 h (room temperature, protected from light) by drip incubation. After washing three times with PBS, cells were incubated for 10 min (room temperature, protected from light) with Hoechst 3342 solution (1:1000) via a dropwise method. Following three more washes with PBS, three-dimensional fluorescence signals of the cells were acquired using the Z-stack module of a confocal fluorescence microscope (Zeiss, LSM880). The fluorescence intensity and volume parameters of different fluorescence channels were analyzed using ImageJ and Imaris software. The results are as follows: Figure 5 As shown.

[0071] (4) Assessment of HMEC-1 cell adhesion function:

[0072] First, peripheral blood was collected from three healthy individuals, and human CD4+ was isolated. + T cells. The method is as follows: ① Lysis of red blood cells: Mix anticoagulated blood with red blood cell lysis buffer (1×) at a ratio of 1:2, let stand for 10 min, then centrifuge (1300 rpm, 10 min, room temperature), discard the supernatant, and repeat the lysis; ② Immunomagnetic bead sorting: Resuspend blood cells in 100 μL of magnetic bead antibody (Miltenyi Biotech, cat.no. 130-045-101) incubation medium, and add an appropriate amount of CD4 magnetic beads (20 μL / 10 ... 7 Cells (Miltenyi Biotech, cat. no. 130-045-101) were incubated for 15 min (4°C, protected from light). Unbound magnetic beads were washed away with magnetic bead antibody incubation solution, and the cells were added to a magnetic column. Finally, the cells bound to the magnetic column, i.e., CD4+, were collected. + T cells.

[0073] Next, CD4 cells were treated with Calcein-AM (1 μmol / L, Invitrogen, cat.no. C1430) live cell dye. + T cells were stained for 15 min (37℃), then centrifuged (300g, 7 min) after adding twice the volume of PBS to wash away excess dye. The supernatant was discarded and the cells were resuspended in fresh RPMI-1640 medium (Gibco, cat. no. 31870082).

[0074] Finally, CD4 + T cells (1×10) 5 CD4 + T cells (per well) were added to 24-well plates containing three groups of HMEC-1 cells and co-cultured for 5 hours. The 24-well plates were then gently washed five times with PBS to remove unattached CD4+ cells. + T cells. Fluorescently labeled CD4+ cells were observed in 24-well plates using an immunofluorescence microscope (Nikon, Eclipse Ti-S). + T cell counts were determined by acquiring images from five different fields of view per well. CD4 counts were performed using ImageJ software. + The number of T cells. The results are as follows: Figure 6 As shown.

[0075] analyze Figure 5 It can be known that:

[0076] Immunofluorescence and three-dimensional reconstruction results showed that treatment of HMEC-1 cells with cell supernatant stimulated by psoriasis-related cytokines reduced the expression levels and cell size of key vascular endothelial barrier molecules hyaluronic acid and heparan sulfate, indicating vascular endothelial barrier damage, with statistically significant differences compared to the control group (P<0.0001). Furthermore, intervention with anti-IGFBP7 neutralizing antibody significantly upregulated the expression of hyaluronic acid and heparan sulfate in HMEC-1 cells, demonstrating that anti-IGFBP7 antibody intervention can significantly improve the structure of the human skin microvascular endothelial barrier and has a protective effect.

[0077] analyze Figure 6 It can be known that:

[0078] Cell co-culture experiments showed that after HMEC-1 cells were treated with cell supernatant stimulated by psoriasis-related cytokines, HMEC-1 cells showed reduced susceptibility to human CD4+. + The number of T cell adhesions was significantly increased, showing a statistically significant difference compared to the control group (P<0.0001). Furthermore, intervention with anti-IGFBP7 antibody significantly inhibited the adhesion of human CD4+ by HMEC-1 cells. + T cell adhesion ability (P<0.0001).

[0079] This indicates that anti-IGFBP7 antibodies play an important role in improving the vascular barrier and reducing psoriasis-like immune inflammation in human cell experiments.

[0080] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the present invention, and all such modifications and substitutions should be covered within the scope of the claims of the present invention. Technical aspects, shapes, and structures not described in detail in this invention are all well-known technologies.

Claims

1. The use of an anti-IGFBP7 antibody in the preparation of a medicament for treating psoriasis by inhibiting IGFBP7 in the blood vessels of the skin, wherein the antibody is selected from the anti-IGFBP7 monoclonal antibody of ABclonal, cat.no. A4615 or the anti-IGFBP7 neutralizing antibody of Sinopharm, cat.no. 13100-MM01.

2. Application of anti-IGFBP7 antibody in the preparation of a drug for treating psoriasis vulgaris, wherein the antibody is selected from the anti-IGFBP7 monoclonal antibody of ABclonal, catalog number cat.no. A4615 or the anti-IGFBP7 neutralizing antibody of Sinopharm, catalog number cat.no. 13100-MM01.

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