Use of C5aR1 in the manufacture of a product for the diagnosis and / or treatment of acute phase MOG AD

Abstract

The application discloses application of C5aR1 in preparation of a product for diagnosing and / or treating acute MOGAD, including application of C5aR1 in preparation of a product for diagnosing acute MOGAD, wherein the positive rate of C5aR1 on the surface of peripheral blood myeloid cells of the patient with acute MOGAD is significantly increased. The application establishes the pathological role of the C5a-C5aR1 axis in the acute attack period of MOGAD, and by blocking the C5aR1 target point on the signal axis, the inflammatory damage of the central nervous system can be reduced. In addition, the application takes C5aR1 and C5a as diagnostic markers of acute MOGAD, and provides objective and quantifiable molecular indexes for clinically distinguishing the acute attack period of MOGAD from the remission period.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and more specifically, to the use of C5aR1 in the preparation of products for the diagnosis and / or treatment of acute MOGAD. Background Technology

[0002] Oligodendrocyte glycoprotein antibody-associated disease (MOGAD) is an inflammatory demyelinating disease of the central nervous system (CNS) characterized by positive plasma MOG-IgG.

[0003] In related technologies, acute-phase treatment of MOGAD relies on glucocorticoids and plasma exchange, while maintenance therapy uses immunosuppressants such as azathioprine, mycophenolate mofetil, and rituximab. However, all of these drugs are non-specific immunosuppressants, and rituximab has a relapse-free survival rate of only about 55% in MOGAD.

[0004] It should be noted that the information disclosed in the background section above is only used to enhance the understanding of the background of this disclosure, and therefore may include information that does not constitute prior art known to those skilled in the art. Summary of the Invention

[0005] The technical objective of this application is to address the above-mentioned shortcomings by providing an application of C5aR1 in the preparation of products for the diagnosis and / or treatment of acute MOGAD. This application establishes the pathological role of the C5a-C5aR1 axis in the acute phase of MOGAD. By blocking the C5aR1 target on this signaling axis, it is possible to effectively inhibit the recruitment of myeloid cells to the central nervous system and regulate the Th1 / Treg immune balance, thereby reducing inflammatory damage to the central nervous system. This provides a targeted strategy for the treatment of MOGAD that differs from existing non-specific immunosuppressive regimens. Furthermore, this application uses C5aR1 and C5a as diagnostic biomarkers for acute MOGAD, providing objective and quantifiable molecular indicators for clinically distinguishing between the acute phase and remission phase of MOGAD. These two diagnostic applications reflect the activation status of the C5a-C5aR1 axis at the receptor and ligand levels, respectively, providing clinicians with a two-dimensional reference for assessing the disease activity of patients.

[0006] To achieve the above objectives, this application provides the following technical solution: According to one aspect of this application, there is provided the use of C5aR1 in the preparation of a product for diagnosing acute MOGAD, wherein the positivity rate of C5aR1 on the surface of peripheral blood myeloid cells in patients with acute MOGAD is significantly increased.

[0007] In some embodiments, the product is a diagnostic kit.

[0008] According to another aspect of this application, there is provided the use of C5a in the preparation of a product for diagnosing acute MOGAD, wherein C5a is a C5aR1 ligand and the concentration of C5a in the plasma of patients with acute MOGAD is significantly elevated.

[0009] In some embodiments, the product is a diagnostic kit.

[0010] According to another aspect of this application, there is provided the use of a C5aR1 antagonist in the preparation of a medicament for treating acute MOGAD, wherein the C5aR1 antagonist is PMX205.

[0011] Compared with existing technologies, the advantages and positive effects of this application are as follows: This application uses C5aR1 and C5a as diagnostic biomarkers for acute MOGAD, providing objective and quantifiable molecular indicators for clinically distinguishing between the acute exacerbation and remission phases of MOGAD. The two diagnostic applications reflect the activation status of the C5a-C5aR1 axis at the receptor and ligand levels, respectively, providing clinicians with a two-dimensional reference for assessing the patient's disease activity.

[0012] This application establishes the pathological role of the C5a-C5aR1 axis in the acute phase of MOGAD. By blocking the C5aR1 target on this signaling axis, it is possible to effectively inhibit the recruitment of myeloid cells to the central nervous system and regulate the Th1 / Treg immune balance, thereby reducing inflammatory damage to the central nervous system. This provides a targeted strategy for the treatment of MOGAD that is different from existing non-specific immunosuppressive regimens.

[0013] Furthermore, in clinical patient samples, this application systematically confirmed the activation characteristics of the C5a-C5aR1 axis in the peripheral blood of patients with acute MOGAD through multidimensional detection methods such as transcriptome sequencing, flow cytometry, and ELISA. These characteristics included a strong correlation between C5aR1 gene expression and inflammation module score, a significant increase in the positive rate of C5aR1 on the surface of myeloid cells, and a significant increase in plasma C5a levels. This provides direct clinical evidence for using C5aR1 as a therapeutic target for MOGAD and fills the gap in the study of the complement pathway in the pathogenesis of MOGAD.

[0014] Furthermore, at the animal model level, this application uses MOG-IgG enhanced MOG. 35-55 The EAE model, which mimics the pathological features mediated by MOGAD antibodies, significantly reduced behavioral symptom scores, decreased spinal cord demyelination area and inflammatory cell infiltration, and reduced the number of myeloid cells and CD4-positive T cells in the central nervous system after PMX205 treatment. This demonstrates that C5aR1 antagonists can effectively alleviate the progression of MOGAD in vivo.

[0015] Furthermore, at the cellular level, this application established an in vitro co-culture system of naïve CD4-positive T cells and bone marrow-derived macrophages, demonstrating that PMX205, by acting on C5aR1 on myeloid cells, can inhibit Th1 cell differentiation and promote Treg cell differentiation, shifting the Th1 / Treg balance towards immune regulation. These in vitro experimental results are consistent with the trends observed in T cell subset changes in animals, elucidating the mechanism of action of C5aR1 antagonists at the cellular level and providing a complete in vitro and in vivo chain of evidence for C5aR1 as a therapeutic target for MOGAD.

[0016] Furthermore, the C5aR1 antagonist treatment strategy proposed in this application differs from existing broad-spectrum immunosuppressants in that it has the characteristics of strong targeting and a clear mechanism of action. It can provide new medication options for MOGAD patients, especially those who do not respond well to or are intolerant of existing treatments. It also provides a theoretical basis for expanding the treatment scope of C5aR1 antagonists to the prevention of relapse during the remission phase of MOGAD. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 A schematic diagram of the experimental design process in an embodiment of this application is shown.

[0019] Figure 2 This application illustrates various cell gating strategies in human peripheral blood according to embodiments of the present application.

[0020] Figure 3 This illustrates the Bulk RNA sequencing and differential expression analysis in an embodiment of this application.

[0021] Figure 4 The different complement pathway expression characteristics are shown in the embodiments of this application.

[0022] Figure 5 This illustrates the correlation between complement-related gene expression and the Inflammatory transcription module in embodiments of this application.

[0023] Figure 6 The results of flow cytometry analysis of peripheral blood myeloid cells from MOGAD patients in this application are shown in the embodiments of this application.

[0024] Figure 7The results of flow cytometry analysis of peripheral blood lymphoid cells from MOGAD patients in this embodiment are shown.

[0025] Figure 8 The results of plasma complement ELISA in MOGAD patients are shown in the embodiments of this application.

[0026] Figure 9 This application illustrates MOG-IgG enhanced MOG in an embodiment of the present application. 35–55 Tissue deposition of MOG-IgG in the EAE model.

[0027] Figure 10 The figures show the plasma C5a levels and brain tissue C5aR1 mRNA expression in each group in the embodiments of this application.

[0028] Figure 11 This application illustrates MOG-IgG enhanced MOG in an embodiment of the present application. 35-55 C5aR1 expression in myeloid cells in the EAE model.

[0029] Figure 12 This application illustrates MOG-IgG enhanced MOG in an embodiment of the present application. 35-55 Schematic diagram of EAE induction and treatment protocol.

[0030] Figure 13 The clinical score curves and morbidity-free survival curves for each group in the embodiments of this application are shown.

[0031] Figure 14 The paper illustrates the pathological and quantitative analysis of spinal cord tissue on day 16 post-immunization in an embodiment of this application.

[0032] Figure 15 This application illustrates an embodiment of MOG-IgG enhanced MOG. 35-55 In the EAE model, membrane attack complex (MAC) deposition is visible around the myelin sheath of the CNS.

[0033] Figure 16 The illustration shows representative immunofluorescence staining of paraffin sections of spinal cord Iba-1 (microglia / macrophages) and Gr-1 (granulocytes) in embodiments of this application.

[0034] Figure 17 The images show representative flow cytometry plots of the CNS (left) and spleen (right) of mice in the control group, model group (MOG-IgG), and PMX205 treatment group in the embodiments of this application.

[0035] Figure 18 Representative flow cytometry plots and statistical results of bone marrow myeloid cells from the control group, model group (MOG-IgG), and PMX205 treatment group in the embodiments of this application are shown.

[0036] Figure 19 The images show CNS lymphocytes and CD4+ cells from the control group, model group (MOG-IgG), and PMX205-treated group in the embodiments of this application. + Representative flow cytometry plots and statistical results of cell subpopulations.

[0037] Figure 20 Naive CD4 is shown in the embodiment of this application. + A schematic diagram of an in vitro co-culture system of T cells and BMDMs.

[0038] Figure 21 The flow cytometry plots and statistical results of each cell in the embodiments of this application are shown. Detailed Implementation

[0039] To better understand the above-mentioned objectives, features, and advantages of this application, the application will be further described below with reference to the accompanying drawings and embodiments. It should be noted that, unless otherwise specified, the embodiments and features described in the embodiments of this application can be combined with each other.

[0040] The present application will be further described below with reference to the accompanying drawings and specific embodiments.

[0041] 1. Research Design and Research Subjects This application enrolled 22 adult MOGAD patients (10 in the acute phase and 12 in remission) (see Table 1). Peripheral blood samples were collected from a hospital in Tianjin. This study was approved by the ethics committee of a university in Tianjin, and all participants signed written informed consent forms.

[0042] Table 1 Clinical characteristics of patients with MOGAD

[0043] Figure 1 A schematic diagram of the experimental design process in an embodiment of this application is shown. For example... Figure 1 As shown in the embodiments of this application, peripheral venous blood was collected from MOGAD patients (n=10 in the acute phase and n=12 in the remission phase). After centrifugation, plasma and PBMCs were separated and then subjected to: (1) ELISA to detect soluble complement components; (2) flow cytometry immunophenotyping analysis; and (3) Bulk RNA sequencing to identify DEGs and enrichment pathways. Through multi-omics data integration and bioinformatics analysis, the peripheral immune dynamics of MOGAD were elucidated.

[0044] 2. Inclusion criteria and stage definition: The inclusion criteria include: (1) age ≥ 18 years; (2) confirmed diagnosis of MOGAD. According to the 2023 International MOGAD Expert Group diagnostic criteria, the following conditions must be met simultaneously: ① core clinical demyelinating events, such as optic neuritis, myelitis, ADEM, etc.; ② MOG-IgG positive (using CBA method for detection): clear positive results do not require additional conditions, weak positive results require AQP4-IgG negative (CBA method) and meet one of the 11 additional imaging conditions; ③ exclusion of other diseases (such as AQP4-IgG positive NMOSD, multiple sclerosis, etc.); (3) blood samples can be collected during the defined acute and / or remission window.

[0045] Exclusion criteria include: (1) presence of comorbidities that may affect systemic complement levels, including active infection / fever, active systemic autoimmune disease or hematologic disease, active malignancy, liver failure or end-stage renal disease; (2) poor quality of sample collection or pretreatment (e.g., severe hemolysis); (3) prior to sampling during an attack, the patient received high-dose glucocorticoid pulse therapy, therapeutic plasma exchange (PLEX) or high-dose intravenous immunoglobulin (IVIG).

[0046] Samples from acute exacerbations were defined as those collected within 30 days of symptom onset; samples from remission periods were defined as those collected 90 days after the last exacerbation, provided there were no recurrences during this period.

[0047] 3. Peripheral blood flow cytometry: Whole blood was collected in EDTA tubes. Sample processing was completed within 6 hours of collection, and flow cytometry analysis was completed within 24 hours. 100 μL of whole blood from each sample was incubated with a mixture of fluorescently labeled antibodies for surface staining. Fc receptors were blocked with anti-CD16 / 32 prior to staining. For markers with low expression or continuous distribution, a fluorescence minus one (FMO) control was used to set the positive threshold. After erythrocyte lysis and PBS washing, data were acquired using a BD FACSAria II flow cytometer and analyzed using FlowJo v10.8.1. Unless otherwise stated, all antibodies were purchased from BioLegend. Granulocytes (CD45+CD11b+CD14- SSC-Ahi), monocytes (CD45+CD11b+CD14+), CD4+... + T cells (CD45+CD3+CD4+) and CD19 + B (CD45+CD3-CD19+).

[0048] Figure 2 This application illustrates various cell gating strategies in human peripheral blood according to embodiments of the present application. Figure 2 A is a gating strategy for flow cytometry used to identify myeloid cell populations in human blood. Figure 2B represents the gating strategy for flow cytometry used for analyzing myeloid cell subsets in human blood.

[0049] 4. Plasma complement ELISA detection Whole blood was collected in EDTA tubes, transported on ice, and processed within 2 hours. Plasma was separated by centrifugation at 4°C (3000 rpm, 10 min), aliquoted into polypropylene cryovials (200 μL per tube), avoiding repeated freeze-thaw cycles (no more than once), and stored at -80°C. Plasma C1q, C5a, and soluble C5b-9 (sC5b-9) levels were quantitatively detected using a commercially available ELISA kit from Lunchangshuo Biotechnology Co., Ltd. All samples were tested in duplicate, and concentrations were calculated using a standard curve fitted with ELISAcalc software.

[0050] 5. PBMC isolation, RNA extraction and transcriptome sequencing PBMCs were isolated from EDTA-anticoagulated blood using density gradient centrifugation, washed twice with PBS, and erythrocyte lysis was performed if necessary. Total RNA was extracted using TRIzol and stored at -80°C for later use. RNA integrity was assessed by agarose gel electrophoresis, and RNA concentration and purity were determined using NanoDrop. Poly(A) was enriched using oligo(dT) magnetic beads. + mRNA was used to construct a library using the VAHTSUniversal V8 RNA-seq Library Prep Kit for Illumina. The library was quantified using Qubit 4.0 and sequenced on the Illumina platform.

[0051] 6. Bioinformatics and statistical analysis of transcriptome data Raw sequencing reads were quality-controlled and filtered using fastp v0.20.1. Clean reads were aligned to the reference genome (GRCh38 / hg38) using STAR v2.7.7a, and gene-level counts were generated using FeatureCounts v2.0.0. Expression quantification was performed using StringTie to obtain raw counts, FPKM, and TPM values. Differential expression analysis was performed using edgeR with TMM normalization and a threshold set to FDR < 0.05 and |log2FC| ≥ 1. Gene Ontology (GO) and KEGG pathway enrichment analyses were performed using clusterProfiler v4.0.2, with Fisher's exact test and Benjamini-Hochberg (BH) correction (after correction). P <0.05 is considered significant.

[0052] The relative proportions of each immune cell subset in PBMCs were estimated using CIBERSORT (LM22 reference matrix, containing 547 characteristic genes of 22 immune cell subsets). Complement-related genes were extracted from the expression matrix, converted to log2(TPM+1), and then Z-score standardized by gene to generate heatmaps. Spearman correlation analysis was used to assess the association between candidate complement genes and inflammation module scores, and BH-FDR correction was performed in each analysis. A multiple linear regression model was constructed to correct for immune cell proportions. Predefined transcriptional modules (inflammation, chemokines, interferons, and antigen presentation) are listed in Table 2. Module scores were calculated using the mean Z-score method: genes within a module were Z-score standardized in each sample, and then the average Z-score within the module was taken.

[0053] Table 2 Predefined transcription modules

[0054] 7. Animal husbandry, experimental protocol design, and ethical approval Female C57BL / 6 mice (4-6 weeks old) were housed in SPF conditions with a stable circadian rhythm of 12 hours light / 12 hours dark, and had free access to food and water. A total of 15 mice were randomly divided into 3 groups (n=5 per group): (1) control group (isotype control + solvent); (2) model group (MOG-IgG + solvent); (3) treatment group (MOG-IgG + PMX205). Experimental procedures were initiated after at least 7 days of acclimatization. Animals were randomly assigned to experimental groups, and all analyses were performed in a blinded manner. All animal experimental protocols were approved by the institution's animal ethics committee (IRB2024-DW-92).

[0055] Constructing MOG-IgG enhanced MOG 35-55 Experimental autoimmune encephalomyelitis (EAE) animal model. EAE was induced by subcutaneous immunization: 200 μg MOG was administered... 35-55 The peptide (GenScript) was emulsified with complete Freund's adjuvant (Sigma) and injected subcutaneously into mice. Pertussis toxin (GLPBIO) was injected intraperitoneally on days 0 and 2. On day 10 post-immunization, mice were intravenously injected with either anti-MOG monoclonal antibody (clone 8-18C5, IgG1 isotype, 200 μg; GenScript) or IgG1 isotype control antibody (200 μg) to establish a MOG-IgG enhanced EAE model.

[0056] The C5aR1 antagonist PMX205 was purchased from MedChemExpress (MCE, catalog number HY-110136, 50 mg) and reconstituted at an appropriate concentration with sterile PBS containing 10% DMSO. It was administered intraperitoneally once daily from days 3 to 16 at a dose of 3 mg / kg (adjusted according to body weight, approximately 45-66 μg per mouse). All mice were scored daily according to the following criteria (0-10 points): 0 = normal; 1 = decreased tail tone; 2 = tail weakness, impaired righting reflex; 3 = absent righting reflex; 4 = gait ataxia; 5 = mild hind limb paraplegia; 6 = moderate paraplegia; 7 = severe paraplegia or hind limb paralysis; 8 = quadriplegia; 9 = near death; 10 = death. The humanitarian endpoint was defined as a score ≥8 points, or a score >7 points for 3 consecutive days. Animals that have reached the humane end of their life are euthanized prematurely, and their final scores are used for subsequent analysis.

[0057] 8. RNA extraction and real-time quantitative PCR from spinal cord tissue Total RNA was extracted from tissues using the TRIzol Reagent kit (GLPBIO, catalog number GK20008). 1 μg of total RNA was used for reverse transcription using the HiScript III All-in-one RT SuperMix Perfect for qPCR (Vazyme, Nanjing) under the following conditions: 50℃ for 15 minutes, 85℃ for 5 seconds, and hold at 4℃. Quantitative PCR was performed in 96-well optical plates using 2×RealStar Fast dye qPCR premix (KangRunChengYe, Beijing), with a total reaction volume of 20 μL. The cycling conditions were: 95℃ for 2 minutes; then 95℃ for 15 seconds, 60℃ for 30 seconds, and 72℃ for 30 seconds; followed by melting curve analysis at 65-95℃. β-actin was used as an internal reference gene, and the relative expression level was calculated using the 2^-ΔΔCt method. Primer sequences are shown in Table 3.

[0058] Table 3. qPCR primer sequences (5′→3′)

[0059] 9. CNS Histopathology and Immunofluorescence In MOG 35-55In the induced EAE model, mice were deeply anesthetized on day 16 post-immunization and perfused with PBS via the heart. Brain and spinal cord (lumbar enlargement) tissues were collected, fixed in 4% paraformaldehyde at 4°C for 24 hours, embedded in paraffin, and prepared into 5 μm thick sections. Inflammatory infiltration was assessed using hematoxylin-eosin (H&E; Solarbio) staining, and demyelination was assessed using Lucker's fast blue (LFB; Solarbio) staining, with semi-quantitative scoring. Inflammatory infiltration score (0–4 points): 0 = no infiltration; 1 = intradural infiltration only; 2 = 1–2 perivascular infiltrations; 3 = 3–4 perivascular infiltrations and / or extensive parenchymal involvement; 4 = large-area infiltration, involving >20% of the area. Demyelination score (0–3 points): 0 points = no demyelination; 1 point = occasional focal demyelination; 2 points = multiple demyelination areas; 3 points = large-area demyelination.

[0060] Antigen retrieval was performed prior to immunofluorescence staining. Primary antibodies included rabbit anti-Iba-1 (1:1000; Wako), rat anti-Gr-1 (1:200; Abcam), rabbit anti-MBP (1:200; Abcam), and anti-C5b-9 (1:100; Santa Cruz Biotechnology). After washing with PBS, sections were incubated with donkey anti-rabbit and donkey anti-rat secondary antibodies, mounted with DAPI-containing anti-fluorescence quenching mounting medium, and imaged using a Nikon fluorescence microscope. Data analysis was performed using ImageJ software.

[0061] 10. Isolation and flow cytometry of immune cells from the CNS, spleen, and bone marrow. Prior to tissue collection, the heart was perfused with ice-cold PBS to minimize contamination by blood-derived immune cells. Brain and spinal cord tissues were minced, digested with type IV collagenase (1 mg / mL, 37°C for 30 minutes), mechanically dissociated, washed, and then demyelinated with 30% Percoll; the resulting cell pellet was collected for flow cytometry. The spleen was mechanically dissociated into a single-cell suspension and filtered through a 70 μm filter. Bone marrow cells were flushed from the femur and tibia. The spleen and bone marrow suspensions were subjected to erythrocyte lysis and washing. Monocytes (CD11b) were also washed. + Ly6C + ), neutrophils (CD11b + Ly6G + ), macrophages (CD11b + F4 / 80 + MHCII+ macrophages (CD11b) + F4 / 80 + MHCII + ).

[0062] For Th1 / Th2 / Th17 cytokine detection, cells were stimulated at 37°C for 6 hours prior to staining using a cell stimulation mixture (ThermoFisher Scientific). Surface staining was performed after Fc blockade. Intracellular IFN-γ, IL-17A, and IL-4 staining was performed using the Intracellular Fixation & Permeabilization Buffer Set (ThermoFisher Scientific), and nuclear Foxp3 staining was performed using the Transcription Factor Buffer Set (BDBiosciences). Positive thresholds for low-expression or continuously distributed markers were set using an FMO control. CD4 + T cells (CD3) + CD4 + CD8 + T cells (CD3) + CD8 + ), B cells (CD45) + CD3 - cd19 + NK cells (CD45) + CD3 - CDNK1.1 + ), Th1 cells (CD3) + CD4 + IFN-γ + ), Treg cells (CD3) + CD25 + FOXP3 + ), Th2 cells (CD3) + CD4 + IL-4 + ), Th17 cells (CD3) + CD4 + IL-17 + Unless otherwise stated, all flow cytometry antibodies were purchased from BioLegend.

[0063] 10. Naïve CD4 + T cells co-cultured with BMDMs and treated with PMX205 in vitro Naïve CD4 was isolated from mouse spleen using a magnetic bead negative sorting method (STEMCELL Technologies). +T cells, with a purity >90%, were pre-activated for 3 days in RPMI 1640 complete medium containing IL-2 (10 ng / mL; ABclonal) with anti-CD3 / CD28 magnetic beads (Thermo Fisher Scientific).

[0064] Bone marrow-derived macrophages (BMDMs) were obtained by differentiating bone marrow cells for 7 days in the presence of M-CSF (100 ng / mL; Sino Biological). Mature BMDMs were stimulated with LPS (100 ng / mL; Solarbio) in combination with IFN-γ (20 ng / mL; Sino Biological) and simultaneously loaded with MOG. 35 – 55 Peptide (20 μg / mL; GenScript) was treated for 24 hours, followed by washing three times with PBS.

[0065] naïve CD4 + T cells were co-cultured with BMDMs in a 96-well plate. The ratio of T cells to BMDMs was 5:1 (T cells: 1 × 10⁻⁶). 5 BMDMs: 2 × 10 4 Cells were cultured in wells (200 μL per well) at 37°C and 5% CO2 for 3–5 days. PMX205 (50 or 100 μg / L) was added at the start of co-culture (t = 0), with a solvent control provided. At the experimental endpoint, cells were collected, anti-CD3 / CD28 magnetic beads were removed magnetically, and Th1 / Treg indices were detected by flow cytometry as described above.

[0066] Experimental results: 1. In PBMCs of patients with acute MOGAD, the expression of inflammation and chemokine-related genes was upregulated, accompanied by changes in the composition of immune cells.

[0067] We performed bulk RNA sequencing and differential expression analysis on peripheral blood mononuclear cells from MOGAD patients in the acute and remission phases. Figure 3 This illustrates the Bulk RNA sequencing and differential expression analysis in an embodiment of this application. Figure 3 A is a volcano diagram of differentially expressed genes between the PBMC transcriptome in the acute phase (n=10) and the remission phase (n=12); Figure 3 B represents the GO biological process enrichment analysis of DEGs; the size of the dots represents gene counts, and the color of the dots reflects -log 10 (Corrected P-value) Figure 3 C represents the KEGG pathway enrichment analysis of DEGs; the size of the dots indicates gene count, and the color of the dots reflects the -log10 (Corrected P-value); Figure 3 D represents the protein-protein interaction (PPI) network of DEGs, showing the inflammation / chemokine-related module and the proliferation / cell cycle module; Figure 3 E represents the proportion of immune cells (22 immune subsets) inferred by CIBERSORT in the acute and remission phases; the comparison between groups was performed using the two-sided Wilcoxon rank-sum test, with P<0.05, P<0.01, P<0.001, and ns indicating no significance.

[0068] like Figure 3 As shown in Figure A, the volcano plot results reveal significant transcriptomic differences in acute-phase samples compared to the remission phase: upregulation of genes such as IL1R1, IL1RL1, ENTPD2, AQP9, FN1, MMP25, OSM, and SLPI, while downregulation of genes such as CXCL9, TMEM176A, DUOX2, and ANOS1. These changes suggest that the transcriptomic characteristics of acute-phase PBMCs are mainly characterized by enhanced inflammatory receptor signaling and increased expression of myeloid cell activation-related molecules.

[0069] like Figure 3 As shown in B, the GO biological process results indicate that differentially expressed genes in the acute phase are mainly enriched in inflammation-related processes such as cytokine-mediated signaling pathways and regulation of inflammatory responses, as well as chemotaxis-related processes such as cell chemotaxis, leukocyte migration, and myeloid leukocyte migration. Figure 3 B).

[0070] like Figure 3 As shown in C, KEGG pathway enrichment analysis further revealed that differentially expressed genes were significantly enriched in multiple immune inflammation and chemokine regulatory pathways, including cytokine-cytokine receptor interaction, chemokine signaling pathway, TNF signaling pathway, IL-17 signaling pathway, NOD-like receptor signaling pathway, and NF-κB signaling pathway.

[0071] like Figure 3 As shown in D, protein-protein interaction network analysis revealed that differentially expressed genes can form two functionally related core modules: one mainly consists of genes related to inflammation response and chemokines, and the other mainly involves cell proliferation and cycle regulation. This suggests that the acute peripheral immune status is not only characterized by the upregulation of inflammation and chemokine-related gene expression, but also by the activation of cell proliferation-related molecular networks.

[0072] like Figure 3 As shown in E, CIBERSORT analysis results showed that the proportion of naive B cells and activated CD4 memory T cells was increased in the acute phase sample, while the proportion of monocytes was decreased, suggesting that the composition of peripheral immune cells may have been redistributed in the acute phase, manifested as enhanced adaptive immune activation, while myeloid cells were reduced in the peripheral blood.

[0073] In summary, during the acute phase of MOGAD, peripheral blood PBMCs exhibit enhanced inflammation and chemotaxis-related transcriptional programs, accompanied by alterations in the proportion of immune cells.

[0074] 2. MOGAD in the acute and remission phases exhibited different complement pathway expression characteristics, among which C5aR1 was associated with the stability of the inflammatory transcriptional module.

[0075] Figure 4 This illustrates different complement pathway expression characteristics in embodiments of this application. Among them, Figure 4 A is a heatmap of complement-related gene expression in PBMCs. The samples (columns) are annotated by disease stage, the genes (rows) are clustered hierarchically, and the expression level is represented by row-wise Z-score (color scale range -2 to +2). Figure 4 B represents the correlation between complement-related gene expression and the NF-κB / TNF transcriptional module; Figure 4 After adjusting for immune cell composition, C represents the multiple linear regression model. The association between C5aR1 expression and inflammatory transcriptional activity remains significant. The dots and horizontal lines represent the regression coefficients (β) and their 95% confidence intervals. The dependent variable is the NF-κB / TNF module, and the covariates include monocytes, naïve B cells, and activated CD4 memory T cells. Figure 4 D represents the Spearman correlation coefficient (r) between C5aR1 expression and multiple inflammatory modules (NF-κB / TNF, chemokines, interferon, and antigen-presenting modules), with the dashed line indicating r = 0. Statistical significance: P <0.05, P <0.01, P <0.001, ns indicates not significant.

[0076] like Figure 4A heatmap of complement-related gene expression revealed that, in PBMCs, genes related to the complement pathway and receptor axis (such as CFH, C4A, C1QA / C1QB / C1QC, C3, C5aR1, C3AR1, CD55, CD59, and ITGAM) exhibited overall expression patterns differing between acute and remission phase samples, along with significant inter-individual heterogeneity. Standardized expression levels (color scale range approximately -2 to 2) showed that in some acute phase samples, C1Q complex genes (C1QA / C1QB / C1QC) and key nodes such as C3, C5aR1, and C3AR1 showed a relatively consistent high expression trend. These genes, along with CD55, CD59, and ITGAM, which are also involved in complement regulation and myeloid-related processes, form a co-expression module, suggesting that the acute phase peripheral immune status may be accompanied by an enhancement of the complement effector axis. In contrast, the expression levels of the above-mentioned genes in the remission period samples were mostly close to the mean, but some individual samples still showed relatively high expression of certain genes (such as CFH), indicating that there was also a certain degree of complement-related transcriptional fluctuation during the remission period.

[0077] In analyzing the association between complement molecules and inflammatory transcriptional programs, we selected candidate complement-related genes (C5aR1, C3AR1, C1QA, ITGAM) and performed Spearman correlation assessments on them with two types of inflammatory modules: one is the "NF-κB / TNF module" (main model), which focuses more on the TNF–NF-κB axis. Figure 4 (B) Another category is the "inflammation module" which encompasses multiple pro-inflammatory cytokines and chemokines (sensitivity analysis, including IL1B, IL6, TNF, CXCL8, CXCL2, CCL2, CCL3, CCL4, NFKBIA, PTGS2, Figure 5 ).

[0078] Figure 5 This paper illustrates the correlation between complement-related gene expression and the inflammatory transcription module in embodiments of this application. The results show that, under both module definitions, C5aR1 is significantly positively correlated with the activity of the inflammatory module, and exhibits the strongest correlation among candidate complement genes: the correlation for the NF-κB / TNF module is r = 0.7561. P = 4.7 × 10 -5 (FDR = 1.88 × 10) -4 The correlation with the inflammation module was r = 0.6488. P= 0.001089 (FDR = 0.004358). In contrast, the other complement-related genes (C3AR1, C1QA, ITGAM) showed weaker overall correlation with the inflammation module score. These results indicate that C5aR1 is the key molecule among the selected complement molecules that is most closely associated with the activity of the inflammatory transcriptional module and whose results are consistent across different module definitions.

[0079] To assess whether the association between C5aR1 and inflammatory transcriptional activity is influenced by differences in immune cell composition, we constructed a multiple linear regression model. This model used the NF-κB / TNF module as the dependent variable and incorporated C5aR1 expression levels, along with CIBERSORT-indicated monocyte, naive B cell, and activated memory CD4+ expression levels. + T cells were included as a covariate for correction ( Figure 4 C). Regression analysis showed that, after simultaneously adjusting for the above cellular components, C5aR1 still maintained a significant positive association with the NF-κB / TNF module (standardized β = 0.719, 95% CI 0.297–1.141). P = 0.002), and the effects of each cellular component covariate in this model did not reach a statistically significant level. These results indicate that the association between C5aR1 and NF-κB / TNF transcriptional program activity is not statistically dependent on the proportion of immune cells, supporting C5aR1 as an independent molecular characteristic reflecting the state of inflammatory transcriptional activation.

[0080] To systematically evaluate the association between C5aR1 and key inflammation-related transcriptional programs, we calculated the Spearman correlations (N = 22) between C5aR1 expression and NF-κB / TNF, chemokines, interferon, and antigen presentation modules. Figure 4 D). The results showed that C5aR1 was significantly positively correlated with the NF-κB / TNF module (r = 0.756, P = 4.7 × 10 -5 FDR = 1.88 × 10 -4 It was also significantly positively correlated with the antigen presentation module (r = 0.608). P = 0.00268, FDR = 0.00535), showing a moderate positive correlation with the chemokine module (r = 0.451, P = 0.0351, FDR = 0.0468). In contrast, although C5aR1 showed a positive correlation with the interferon module (r = 0.371), it did not reach statistical significance. P= 0.0892, FDR = 0.0892). In summary, the immune activation state associated with C5aR1 is mainly characterized by inflammation amplification driven by the TNF–NF-κB axis, accompanied by a synchronous enhancement of chemokine production and antigen presentation.

[0081] 3. Flow cytometry showed that the proportion of peripheral blood myeloid cells changed during the acute phase of MOGAD, accompanied by an increased positive rate of C5aR1 in myeloid cells.

[0082] Figure 6 The results of peripheral blood flow cytometry in MOGAD patients are shown in the embodiments of this application. Figure 6 A represents a change in the frequency of peripheral blood myeloid cells; Figure 6 B represents C5aR1 expression in peripheral blood myeloid cells. For example... Figure 6 As shown in Figure A, compared to the remission phase, the composition of myeloid cells changes during the acute phase, mainly manifested as CD11b... + The proportion of cells increased, the proportion of granulocytes increased, while the proportion of monocytes decreased. For example... Figure 6 Further analysis of receptor phenotypes in myeloid subsets revealed that CD11b in the acute phase... + The C5aR1 positivity rates of myeloid cells, monocytes, and CD86+ monocytes were all higher in the acute phase than in the remission phase. These results indicate that myeloid cells not only undergo changes in their numerical composition during the acute phase but also exhibit higher levels of C5aR1 expression.

[0083] Figure 7 The results of peripheral blood flow cytometry in MOGAD patients are shown in the embodiments of this application. Figure 7 As shown, in terms of lymphoid cells, CD4 in acute phase samples + T cells and CD19 + The proportion of B cells was increased, suggesting that the peripheral immune pattern in the acute phase showed changes in adaptive immune components.

[0084] 4. In the acute phase of MOGAD, plasma C5a and C1q levels are elevated, while sC5b-9 remains stable.

[0085] After observing changes in cell phenotype, we measured the levels of complement-related components in patient plasma to further assess the overall status of circulating complement. Figure 8 The results of plasma complement ELISA in MOGAD patients are shown in the embodiments of this application. Figure 8As shown, compared with the remission phase, the plasma levels of C1q and C5a were significantly elevated in patients during the acute phase. Conversely, the marker sC5b-9, reflecting the terminal complement activation process, showed no significant difference between the acute and remission phases. In summary, changes in circulating complement in patients during the acute phase mainly manifested as a decrease in upstream components (C1q) and their cleavage products (C5a), without the increased formation of terminal complement complexes suggested by elevated sC5b-9 levels.

[0086] 5. MOG-IgG Enhanced MOG 35–55 EAE model establishment and antibody deposition validation.

[0087] Figure 9 This application illustrates MOG-IgG enhanced MOG in an embodiment of the present application. 35–55 Tissue deposition of MOG-IgG in the EAE model. For example... Figure 9 As shown, in situ IgG deposition was observed in the MOG-IgG group after staining with anti-mouse secondary antibody (cell nuclei were counterstained with DAPI). Scale bar, 20 μm. Control group (left) and MOG-IgG group (right).

[0088] 6. C5aR1 expression is upregulated in the central nervous system tissue of MOG-IgG enhanced EAE.

[0089] Figure 10 The expression of C5aR1 mRNA in each group is shown in the embodiments of this application. Among them, Figure 10 A indicates that the plasma C5a level in the MOG-IgG group was significantly higher than that in the control group; Figure 10 B represents the expression of C5aR1 mRNA in spinal cord tissues of the control group and the MOG-IgG group. Significance is indicated by asterisks (*<0.05, **<0.01, ***<0.001, ****<0.0001). qPCR detection showed that C5aR1 expression in CNS tissues of the MOG-IgG group was significantly higher than that of the control group. Figure 10 A), while plasma free C5a was also significantly increased ( Figure 10 B).

[0090] Figure 11 This application illustrates MOG-IgG enhanced MOG in an embodiment of the present application. 35-55 C5aR1 expression in myeloid cells in the EAE model. Figure 11 A represents the proportion of macrophages in the bone marrow, spleen, and central nervous system; Figure 11B represents the expression ratio of C5aR1 in pro-inflammatory and anti-inflammatory macrophages in the bone marrow, spleen, and central nervous system; data are expressed as mean ± SD, and significance is indicated by an asterisk (*<0.05, **<0.01, ***<0.001, ****<0.0001). Flow cytometry further showed that in myeloid cells, CNS macrophages had the highest C5aR1 expression, followed by the spleen, and the lowest expression in the bone marrow. Figure 11 A). In addition, pro-inflammatory macrophages (MHCII) + C5aR1 expression in the anti-inflammatory type (CD206) was higher in all tissues than in the anti-inflammatory type (CD206). + () Figure 11 (B) This suggests that C5aR1 is closely related to the pro-inflammatory myeloid phenotype. These results indicate that C5aR1 is predominantly expressed in the CNS and inflammatory myeloid cells in this model, providing a basis for PMX205 intervention studies.

[0091] 7. PMX205 intervention delays disease onset and reduces symptom severity.

[0092] In MOG-IgG enhanced MOG 35 – 55 In the EAE model, PMX205 was administered after immunization. Figure 12 This application illustrates MOG-IgG enhanced MOG in an embodiment of the present application. 35-55 Schematic diagram of EAE induction and treatment protocols. Under the same conditions, the EAE clinical scoring system was used to dynamically assess disease progression.

[0093] Figure 13 The clinical score curves and morbidity-free survival curves for each group in the embodiments of this application are shown. Figure 13 A represents the longitudinal EAE symptom score, using a 10-point neurological disability scale to assess the degree of neurological disability in mice. Figure 13 B is the disease-free survival curve (Kaplan-Meier method), showing the morbidity analysis of the two groups of animals. Figure 13 Data A is expressed as mean ± SD. P <0.05, P <0.01, P <0.001, **** P <0.0001. Compared with the untreated model group, PMX205 treatment delayed disease onset (approximately 2–3 days) and reduced overall disease severity, as evidenced by a downward shift and reduced peak value of the clinical score curve. Figure 13 A). This suggests that C5aR1 blockade can alleviate neurological dysfunction during disease activity. Survival analysis showed that PMX205 had a limited impact on overall morbidity. Figure 13(B) This means that the morbidity rate of the animals was not significantly altered during the observation period. In summary, the ameliorative effect of PMX205 in this model is mainly manifested in delaying the onset of disease and reducing the severity of symptoms.

[0094] 8. PMX205 can effectively reduce inflammatory cell infiltration and demyelination damage in the central nervous system.

[0095] Figure 14 This illustration shows the histopathological and quantitative analysis of spinal cord tissue on day 16 post-immunization in this application embodiment. HE staining (inflammatory infiltration), LFB staining (demyelination), and MBP immunofluorescence staining (myelin integrity) are included. Quantitative data are shown in the side bar chart, and the scale bar is shown in the figure. Data are expressed as mean ± SD. P <0.05, P <0.01, P <0.001, **** P <0.0001.

[0096] To assess the impact of C5aR1 blockade on central nervous system pathological damage, CNS tissues were stained with HE, LFB, and MBP. Figure 14 Compared with the MOG-IgG group, the PMX205 group showed reduced inflammatory infiltration, a smaller demyelinated area, and enhanced MBP signal, indicating reduced myelin damage. These results suggest that PMX205 can alleviate inflammatory infiltration and demyelination pathology at the histological level.

[0097] Figure 15 This application illustrates an embodiment of MOG-IgG enhanced MOG. 35-55 In the EAE model, membrane attack complex (MAC) deposition is visible around the myelin sheath of the CNS. Figure 15 Representative immunofluorescence staining of spinal cord sections from mice in the control group, model group (8-18C5 MOG-IgG), and PMX205 treatment group is shown. The sections were stained with C5b-9 (MAC; red) and myelin basic protein (MBP; green), and the combined image is shown. The lesion group showed increased C5b-9 signal and increased signal intensity with MBP. + Co-localization. Scale bar, 25 μm. Immunofluorescence staining of CNS tissues showed clear MAC deposition signals in the model group. Figure 15 The results indicate that the model exhibits activation of the complement terminal pathway and MAC deposition.

[0098] 9. PMX205 intervention reduces central myeloid cell infiltration accompanied by a relative increase in peripheral splenic myeloid cells.

[0099] To further evaluate the effect of C5aR1 blockade on central inflammatory infiltration, immunofluorescence and flow cytometry were used to perform multi-level verification of central myeloid cell infiltration in the MOG-IgG enhanced EAE model.

[0100] Figure 16 The illustration shows representative immunofluorescence staining of paraffin sections of spinal cord Iba-1 (microglia / macrophages) and Gr-1 (granulocytes) in embodiments of this application. Scale bar, 40 μm. Data are expressed as mean ± SD, and significance is indicated by an asterisk (*<0.05, **<0.01, ***<0.001, ****<0.0001).

[0101] Figure 16 Immunofluorescence results showed that, compared with the MOG-IgG group, the PMX205 group had lower levels of Iba-1 in the CNS. + Cells and Gr-1 + Reduced cell infiltration suggests that C5aR1 blockade can effectively inhibit myeloid cell infiltration in the CNS.

[0102] We used flow cytometry to quantitatively verify the above findings. Figure 17 The images show representative flow cytometry plots of the CNS (left) and spleen (right) in mice from the control group, MOG-IgG group, and PMX205 group in this application. Frequencies are expressed as CD45. + Percentage of cells. Data are expressed as mean ± SD, and significance is indicated by an asterisk (*<0.05, **<0.01, ***<0.001, ****<0.0001).

[0103] Figure 17 The results showed that in CNS tissues, CD11b levels decreased after PMX205 intervention. + The overall proportion of myeloid cells decreased, and several key myeloid subsets (including monocytes, neutrophils, macrophages, and MHCII+ macrophages) showed a consistent decreasing trend. This result is corroborated by immunofluorescence observations, further confirming that PMX205 can effectively inhibit the infiltration of central myeloid cells at the cell population level.

[0104] Notably, contrary to the central nervous system, in the peripheral spleen, CD11b levels were higher in the PMX205 intervention group. + The proportions of cells, monocytes, neutrophils, and macrophages were all relatively increased. Figure 17 This distribution pattern of "central decline and peripheral rise" suggests that C5aR1 blockade may affect the dynamic migration of myeloid cells between the periphery and the center, that is, the recruitment process of peripheral myeloid cells to the center is inhibited, resulting in retention or accumulation in the periphery.

[0105] To rule out the potential confounding factor of differences in myelogenesis at the bone marrow ends, we further examined the composition of myeloid cells in the bone marrow. Figure 18 Representative flow cytometry plots and statistical results of bone marrow myeloid cells from the control group, MOG-IgG group, and PMX205 group in this application are shown. Data are expressed as mean ± SD, and significance is indicated by an asterisk (*<0.05, **<0.01, ***<0.001, ****<0.0001). Figure 18 As shown, in MOG-IgG enhanced MOG 35-55 In the EAE model, the proportion of myeloid cells in the bone marrow increased, but PMX205 treatment did not change this. After PMX205 intervention, CD11b levels in the bone marrow... + The proportions of cells, neutrophils, monocytes, and macrophages did not differ significantly between groups. This suggests that the reduction in central myeloid cell infiltration is more likely due to impaired peripheral-to-central migration rather than being driven by differences in bone marrow myelogenesis.

[0106] 10. PMX205 intervention can reduce central CD4 levels. + T cell infiltration causes a shift in the Th1 / Treg immune balance: manifested as a decrease in the proportion of Th1 cells and an increase in the proportion of Treg cells.

[0107] To assess the effect of C5aR1 blockade on CNS lymphocyte infiltration, we used flow cytometry to analyze CNS infiltrating lymphocytes. Figure 19 The images show CNS lymphocytes and CD4+ cells from the control group, MOG-IgG group, and PMX205 group in the embodiments of this application. + Representative flow cytometry plots and statistical results for cell subsets. Data are expressed as mean ± SD, and significance is indicated by an asterisk (*<0.05, **<0.01, ***<0.001, ****<0.0001).

[0108] Figure 19 The results showed that, compared with the MOG-IgG group, CD4 in the CNS was significantly reduced after PMX205 treatment. + Decreased T cell frequency suggests that C5aR1 blockade can alleviate central CD4+. + T-cell infiltration.

[0109] Further analysis of CD4 in CNS + Functional subset analysis of T cells revealed that after PMX205 intervention, Th1 (IFN-γ) + The proportion of Treg (CD25) cells decreased, while the proportion of Treg (CD25) cells decreased. + FOXP3 +The proportion of CD4+ cells increased. These results indicate that C5aR1 blockade not only reduced the number of infiltrating central lymphocytes but also was accompanied by an increase in CD4+ cells. + The functional shift of T cells, namely the tilt from pro-inflammatory Th1 responses to regulatory Treg responses, manifests as a shift in the Th1 / Treg balance toward immune regulation.

[0110] Based on the association between C5aR1 and antigen-presenting-related transcriptional procedures observed in previous transcriptome analyses, this study further hypothesizes that C5aR1 blockade induces CD4... + Decreased T cell infiltration and Th1 / Treg polarization shift may be related to myeloid cell-mediated antigen presentation and alterations in the inflammatory microenvironment. Based on this hypothesis, we established BMDM–CD4 in subsequent experiments. + A T-cell co-culture system was developed to validate the myeloid-T-cell interaction mechanism in vitro.

[0111] 11. In vitro co-culture experiments confirmed that PMX205 can effectively inhibit Th1 cell differentiation while promoting Treg cell differentiation.

[0112] To investigate the function of C5aR1 signaling in myeloid-T cell interactions and to verify the effect of its pharmacological blockade on T cell differentiation outcomes, we established an naïve CD4+ signaling pathway. + T cell co-culture system with BMDM. In this system, BMDM is first stimulated with LPS combined with IFN-γ and loaded with MOG. 35 – 55 Peptides, mimicking the inflammatory antigen presentation environment, are then pre-activated CD4 by CD3 / CD28. + T cells were co-cultured at a fixed ratio. At the start of co-culture, low-dose and high-dose PMX205 were added for intervention. After co-culture, CD4 counts were analyzed by flow cytometry. + T cell subsets, with a focus on assessing changes in the ratio of Th1 to Treg cells. Figure 20 ).

[0113] Figure 20 Naive CD4 is shown in the embodiment of this application. + A schematic diagram of an in vitro co-culture system of T cells and BMDMs. Figure 21 The flow cytometry plots and statistical results of each cell in the embodiments of this application are shown. Figure 21 A is for displaying CD4 + IFN-γ + Cells and CD4 + CD25 + FOXP3 + Representative flow cytometry plot of gated regulatory T cells, showing percentages of CD4+. +Frequency within the T cell gate; Figure 21 B is CD4 + IFN-γ + Cells and CD4 + CD25 + FOXP3 + Statistical results for Tregs. Data are expressed as mean ± SD, and significance is indicated by an asterisk (*<0.05, **<0.01, ***<0.001, ****<0.0001).

[0114] Figure 21 The results showed that T cell differentiation in the co-culture system exhibited a significant effector bias under inflammatory stimulation and antigen load conditions. After PMX205 intervention, the proportion of Th1 cells decreased and the proportion of Treg cells increased, and this trend was observed in both low- and high-dose groups. Compared with the untreated control group, PMX205 treatment shifted the Th1 / Treg balance from pro-inflammatory to immunomodulatory, suggesting that C5aR1 signaling is involved in regulating inflammatory myeloid cell-mediated CD4+. + T cell differentiation.

[0115] The above in vitro results are consistent with the direction of Th1 / Treg balance changes observed in animal experiments, further confirming under controlled cell-cell interaction conditions that the C5aR1 signaling in myeloid cells is a key node regulating the Th1 / Treg balance. PMX205 can inhibit the Th1 response and promote Treg differentiation by blocking this signaling axis.

[0116] Experimental conclusion: Experimental results showed that the C5a–C5aR1 axis was activated during the acute phase of MOGAD. Inhibiting C5aR1 alleviated disease progression in two ways: first, by reducing the infiltration of myeloid cells such as monocytes and neutrophils into the CNS; second, by regulating the function of antigen-presenting cells and affecting CD4+. + The differentiation direction of T cells can suppress pro-inflammatory Th1 responses and promote Treg cells, thereby regulating the immune response. Therefore, targeting the C5a–C5aR1 axis is a potential therapeutic strategy for MOGAD.

[0117] Through the above specific embodiments, those skilled in the art can easily implement this application. However, it should be understood that this application is not limited to the specific embodiments described above. Based on the disclosed embodiments, those skilled in the art can arbitrarily combine different technical features to achieve different technical solutions.

Claims

1. The application of C5aR1 in the preparation of products for diagnosing acute MOGAD, characterized in that, The positive rate of C5aR1 on the surface of peripheral blood myeloid cells was significantly increased in patients with acute MOGAD.

2. The application according to claim 1, characterized in that, The product in question is a diagnostic reagent kit.

3. The application of C5a in the preparation of products for diagnosing acute MOGAD, characterized in that, The C5a is a C5aR1 ligand, and the concentration of C5a in the plasma of patients with acute MOGAD is significantly elevated.

4. The application according to claim 3, characterized in that, The product in question is a diagnostic reagent kit.

5. The use of C5aR1 antagonists in the preparation of medicaments for the treatment of acute MOGAD, characterized in that, The C5aR1 antagonist is PMX205.

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