Use of an agent in the manufacture of a product for detecting or treating IgG4-RD

Abstract

The application belongs to the field of proteomics research, and discloses application of a reagent in preparation of a product for detecting or treating IgG4-RD. Six PD-L1 related DEPs (RPS6KB1, NFATC1, CD3E, PTEN, PTPN6 and JAK2 proteins) can be used as biomarkers for patient stratification and identification of individuals with high risk of severe disease progression or fibrosis sequelae, secondly, activation of the PD-1 / PD-L1 pathway suggests that immune checkpoint regulation may be a new treatment method for IgG4-RD, thirdly, significant enrichment of metabolic pathways indicates that IgG4-RD is a metabolic disorder disease, which provides a way for targeted immunometabolism. Finally, drug repositioning research highlights resveratrol and sorafenib as candidate therapeutic drugs, and the potential combination of these drugs provides a new method for rebalancing immune activation and inhibition in IgG4-RD.

Description

Technical Field

[0001] This invention belongs to the field of proteomics research, specifically relating to the application of a reagent in the preparation of products for the detection or treatment of IgG4-RD. Background Technology

[0002] IgG4-related disease (IgG4-RD) is a chronic, immune-mediated fibro-inflammatory disease characterized by neoplastic lesions, striae-like fibrosis, and dense lymphoplasmacytic infiltration rich in IgG4-positive plasma cells. This rare systemic fibrotic inflammatory disease has an estimated global prevalence of approximately 5 cases per 100,000 people, primarily affecting middle-aged and older men. IgG4-RD can affect almost any organ—most commonly the pancreas, salivary and lacrimal glands, lymph nodes, and retroperitoneum—and is characterized by organ enlargement or malignant or infectious mass-like lesions, progressive fibrosis that can lead to irreversible tissue damage, and multi-organ involvement that significantly increases morbidity and long-term complications. However, despite increasing clinical understanding, the molecular and immunological mechanisms of organ tropism, chronic inflammatory processes, and fibrosis remain not fully elucidated.

[0003] Immune checkpoint pathways, particularly the PD-1 / PD-L1 axis, play a crucial role in maintaining peripheral tolerance by preventing excessive T cell activation and autoimmunity. Increasing evidence suggests that PD-1 / PD-L1 dysregulation is involved in various autoimmune and inflammatory diseases. In IgG4-RD, patients frequently exhibit aberrant T and B cell activation, excessive plasmablast expansion, and features indicative of immune exhaustion, pointing to a possible role for checkpoint signaling in disease pathogenesis. Studies have shown increased PD-1 expression on peripheral circulating T cells and increased PD-L1 expression in affected tissues. Therefore, checkpoint signaling pathways may be involved in maintaining the chronic immune activation and paradoxical immunosuppressive environment frequently observed in IgG4-RD patients. However, the precise regulatory link between PD-1 / PD-L1 and chronic inflammation, metabolic reprogramming, and fibrosis in IgG4-RD remains largely unclear.

[0004] Studying the plasma proteome is challenging due to the extremely wide dynamic range of circulating proteins. High-abundance proteins, accounting for over 99% of total plasma protein mass, create a "masking effect," preventing the detection of low-abundance proteins using conventional techniques. Therefore, most research has focused on high-abundance proteins, while low-abundance secretions have been relatively underexplored. Despite their low concentrations, these low-abundance proteins often play important roles in early pathophysiological processes and / or disease progression. Therefore, comprehensive characterization of low-abundance plasma proteins is essential for improving our understanding of the molecular mechanisms of IgG4-RD.

[0005] Recent advances in multi-omics technologies have provided new opportunities for the discovery of non-invasive biomarkers. Despite continuous methodological advancements, no research has yet systematically integrated KEGG and Metascape enrichment analysis, PPI networks, pivotal protein assessment, and drug repositioning prediction to identify and evaluate differentially expressed proteins (DEPs), and further validated key PD-1 / PD-L1-related candidate proteins using ELISA to determine the precise regulatory links between PD-1 / PD-L1 and chronic inflammation, metabolic reprogramming, and fibrosis in IgG4-RD, thereby achieving preventive and therapeutic effects. Summary of the Invention

[0006] The purpose of this invention is to overcome at least one deficiency of the prior art and to provide an application of a reagent in the preparation of products for the detection or treatment of IgG4-RD.

[0007] The technical solution adopted in this invention is: In a first aspect, the present invention provides the use of a reagent in the preparation of a product for detecting IgG4-RD, said reagent being a reagent for detecting PD-1 / PD-L1 axis marker levels.

[0008] In some instances, the PD-1 / PD-L1 axis markers are NFATC1 and PTEN proteins.

[0009] In some instances, the PD-1 / PD-L1 axis markers are RPS6KB1, NFATC1, CD3E, PTEN, PTPN6, and JAK2 proteins.

[0010] In some instances, the product is selected from reagent kits, protein chips, test strips, or immunoassay devices.

[0011] In some instances, the immune detection device includes: A quantitative system for detecting PD-1 / PD-L1 axis marker levels in quantitative samples; The analysis system uses a quantitative device to detect the levels of PD-1 / PD-L1 axis markers and compares them with normal expression levels. The result output system is used to output the analysis results of the analysis device.

[0012] In some instances, higher-than-normal expression levels of JAK2, PTEN, and PTPN6, and lower-than-normal expression levels of RPS6KB1, NFATC1, and CD3E indicate IgG4-RD.

[0013] In some instances, the sample is a peripheral blood sample from the subject.

[0014] Secondly, the present invention provides the use of a reagent in the preparation of a product for treating IgG4-RD, said reagent being a PD-1 / PD-L1 axis modulator, inhibitor, or agonist.

[0015] In some instances, the PD-1 / PD-L1 axis regulator, inhibitor, or agonist is a regulator, inhibitor, or agonist targeting the proteins RPS6KB1, NFATC1, CD3E, PTEN, PTPN6, and JAK2.

[0016] In some instances, the PD-1 / PD-L1 axis modulator, inhibitor, or agonist is selected from at least one of resveratrol or sorafenib.

[0017] The beneficial effects of this invention are: This invention demonstrates how immune checkpoint activation, T-cell suppression, and immune-metabolic reprogramming converge to form the basis of disease pathogenesis. Through KEGG, GO, GSEA, and PPI analyses, six DEPs (RPS6KB1, NFATC1, CD3E, PTEN, PTPN6, and JAK2) were identified as core regulators of this pathway, providing unique mechanistic insights into the coexistence of persistent inflammation and immune exhaustion in IgG4-RD.

[0018] Mechanistically, downregulation of RPS6KB1 and NFATC1 reduces T cell metabolic signaling and cytokine transcription, while decreased CD3E levels inhibit TCR-mediated activation. Overall, these changes impair T cell effector function, indirectly promoting PD-1 / PD-L1-mediated repression. Conversely, upregulated JAK2 may promote IL-6 and interferon receptor signaling and PD-1 transcription via the JAK / STAT axis. Upregulated PTEN inhibits PI3K-AKT-mTOR signaling, reducing T cell metabolic adaptability, while simultaneously promoting PTPN6 to directly inhibit TCR signaling, thereby promoting PD-1-mediated repression.

[0019] This invention further validated the findings quantitatively using ELISA, revealing a significant increase in PTEN and a significant decrease in NFATC1 in IgG4-RD patients compared to healthy controls. This supports the dysregulation of the PI3K-Akt and TCR-NFAT axes, which are associated with PD-1 / PD-L1-mediated T cell suppression. In summary, these signaling changes lead to reduced T cell activation and enhanced checkpoint signaling, resulting in an immunosuppressive microenvironment in IgG4-RD. At the energy metabolism level, key regulatory pathways associated with lymphocyte activation were upregulated: mTORC1 signaling, PI3K-AKT-mTOR signaling, and oxidative phosphorylation. Furthermore, enrichment of the epithelial-mesenchymal transition pathway suggests active tissue remodeling and fibrosis. This pathway analysis highlights the dual nature of the IgG4-RD state: chronic inflammation and immunosuppression mediated by checkpoint mechanisms.

[0020] This invention has multiple clinical implications: First, the six PD-L1-related DEPs can serve as biomarkers for patient stratification and identifying individuals at high risk of severe disease progression or sequelae of fibrosis. Second, activation of the PD-1 / PD-L1 pathway suggests that immune checkpoint modulation may be a novel treatment for IgG4-RD, although the effects may be complex, requiring a balance between immune restoration and potential inflammatory exacerbation. Third, significant enrichment of metabolic pathways indicates that IgG4-RD is a metabolic disorder, providing a pathway for targeting immune metabolism. Finally, drug repositioning studies highlight resveratrol and sorafenib as candidate treatments. Resveratrol may reduce PD-L1 expression and restore T cell activity by decreasing JAK2 / STAT3 and NF-kappaB signaling, while sorafenib may modulate the immune system while providing anti-fibrotic properties by inhibiting kinase-driven signaling and the PI3K-AKT-mTOR pathway. The potential of combining these drugs offers a novel approach to rebalancing immune activation and suppression in IgG4-RD. Attached Figure Description

[0021] Figure 1 The Venn diagram shows the intersection between differentially expressed proteins (DEPs) and proteins involved in PD-L1 expression and the PD-1 checkpoint pathway.

[0022] Figure 2 This is a volcano plot that identifies differentially expressed candidate proteins associated with the PD-1 / PD-L1 pathway.

[0023] Figure 3 It is a protein-protein interaction (PPI) network constructed based on differentially expressed proteins.

[0024] Figure 4 These are key core node proteins identified from the PPI network (analyzed using the CytoHubba plugin).

[0025] Figure 5 These are feature modules extracted from the PPI network (analyzed using the MCODE plugin). Figure 6 This shows the differential expression of NFATC1 and PTEN between IgG4-RD patients and healthy controls. Detailed Implementation

[0026] The following disclosure provides many different implementations or examples for different ways of implementing the present invention. Example

[0027] 1. Research Materials and Methods 1.1 Research Cohort The study included 5 patients diagnosed with IgG4-RD and 10 age-matched healthy controls. Inclusion criteria for IgG4-RD were based on the comprehensive diagnostic criteria (CDC 2011) and its 2020 revision (RCD). Patients who were pregnant or lactating, had a history of malignancy, severe hepatic or renal impairment, acute infection, autoimmune disease, or had recently received glucocorticoid or other immunosuppressive therapy were excluded. All participants donated peripheral blood for analysis. The protocol was approved by the ethics committee, and written consent was obtained from each patient.

[0028] Whole blood was collected using K2-EDTA tubes for plasma separation, and coagulant tubes were used for serum separation. EDTA-treated samples were centrifuged at 2200×g for 15 minutes after being left at room temperature, and the plasma supernatant was collected. Plasma and serum were aliquoted (200 μL) and stored at -80°C until proteomics analysis.

[0029] 1.2 Sample Preparation SOMAscan assays employ a proprietary multi-level dilution protocol to improve the detectability of low-abundance proteins. Samples are diluted to three levels: 0.005%, 1%, and 40%, representing high, medium, and low dilutions, respectively. Each SOMAmer reagent is assigned to a level based on the expected endogenous concentration of its respective protein.

[0030] 1.3 SomaScan P11K proteomics analysis SomaScan P11K proteomics assay is a high-throughput aptamer platform for deep, highly specific, and accurate protein quantification. This assay utilizes chemically modified aptamers (called SOMAmer reagents) that exhibit high affinity for target proteins via SELEX screening. Plasma samples are incubated with SOMAmer reagents to form aptamer-protein complexes. After washing to remove unbound proteins, the complexes are released via photolysis. The aptamers are then separated from the proteins, and blocking materials reduce non-specific binding. The aptamers are captured, enriched, and quantified using a fluorescent microarray, with fluorescence signal intensity measuring the amount of raw protein. This platform features a wide dynamic range (approximately 10 orders of magnitude) and high reproducibility (CV 5-7%). A stratified dilution strategy enables accurate detection of both high-abundance and low-abundance proteins in a single assay.

[0031] Proteomics data were acquired using the SomaScan P11K platform (v5.0). Protein identification was performed by mapping SOMAmer reagents to human protein sequences in the UniProt database. Data processing employed standardized normalization procedures: hybridization normalization, median signal normalization, inter-plate scaling, and inter-plate calibration. Quality control used internal reference standards, with a median coefficient of variation (CV) of 4.5%. Raw data underwent multiple normalization. All quantitative measurements were log2 transformed and centered using R software (v4.2.0).

[0032] 1.4 Enrichment Analysis DEPs were functionally annotated using the UniProt database. Subcellular distribution was analyzed using UniProt localization data to understand the distribution of DEPs within cellular compartments. Sequence alignment was performed using the euKaryotic ortholog (KOG) database to classify DEPs into functionally similar categories. Functional and pathway enrichment analyses were performed using the KEGG Automated Annotation Server (KAAS) and the KEGG database. Gene ontology (GO) enrichment analysis was also performed to assess biological processes associated with DEPs.

[0033] 1.5 Construction and identification of protein-protein interaction (PPI) networks and subnetworks DEPs were uploaded to the STRING platform (confidence score threshold 0.4) to construct a PPI network. Network visualization and topology analysis were performed using Cytoscape. Enrichment and module integration analysis were conducted using the Metascape platform to identify closely related functional clusters.

[0034] 1.6 Identification of pivot proteins Hub proteins were identified from the PPI network using the CytoHubba plugin in Cytoscape. The maximum clique centrality (Degree) algorithm scored each node based on its topological importance. The top 15 proteins with the highest Degree scores were selected as hub proteins, representing key regulatory nodes that may be important in the pathogenesis of IgG4-RD.

[0035] 1.7 Enzyme-linked immunosorbent assay (ELISA) Plasma samples from IgG4-RD patients and healthy controls were analyzed by ELISA according to the manufacturer's instructions. PTEN levels were measured using the PTEN ELISA kit (Abcam, catalog number ab206979), and NFATC1 levels were quantified using the Human NFAT2 SimpleStep ELISA kit (NFATC1) (Abcam, catalog number ab277704).

[0036] 1.8 Drug Prediction Enrichment analysis using the Enrichr tool across the drug signature database (DSigDBv1.0) was performed to identify potential novel compounds. A list of target genes was input into Enrichr, and overlap with the drug-related gene set in DSigDB was calculated. Compounds were ranked based on combinatorial enrichment scores and corrected p-values.

[0037] 1.9 Statistical Analysis DEPs were determined using a composite threshold of statistical significance and fold change. Proteins with a fold change (FC) greater than 1.5 or less than 0.67 and an FDR-corrected p-value < 0.05 were considered significant. Pathway enrichment was tested using Fisher's exact test. Statistical significance was defined as: P < 0.05, *P < 0.01, **P < 0.001, ***P < 0.0001.

[0038] 2. Results Analysis 2.1 Potential activation of PD-L1 signaling and metabolic pathways in IgG4-RD patients This study included 5 patients with IgG4-RD and 10 healthy controls, none of whom had evidence of autoimmune diseases other than IgG4-RD. Inclusion criteria for IgG4-RD were based on the CDC (2011) and RCD (2020) criteria, covering clinical, laboratory, and histopathological findings. Exclusion criteria included pregnancy / lactation, history of malignancy, severe liver / kidney failure, acute infection, recent immunosuppressant / glucocorticoid use, or other autoimmune diseases.

[0039] The SomaScan P11K platform systematically investigated molecular changes in the plasma proteome of IgG4-RD. The workflow included plasma sample collection / preparation (n=5 IgG4-RD, n=10 healthy controls) and high-throughput proteomics analysis. Following rigorous quality assessment and data analysis, 248 differentially expressed proteins (DEPs) were identified, representing approximately 2.2% of all quantified proteins. Of these, 180 (72.0%) were significantly upregulated in IgG4-RD patients compared to healthy controls, and 68 (28.0%) were significantly downregulated.

[0040] Functional annotation revealed that DEPs were significantly enriched in cell signaling, nuclear function, and cytoskeleton pathways. Subcellular localization analysis showed that DEPs were mainly distributed in the cytoplasm, nucleus, and cell membrane, suggesting widespread perturbation of cellular processes. KOG analysis classified DEPs into functional categories, with the largest groups being "signal transduction mechanisms," "post-translational modifications, protein turnover, molecular chaperones," and "cytoskeleton."

[0041] To explore the biological significance of DEPs, enrichment analysis was performed using the Metascape platform. The results showed that these DEPs are mainly involved in several key biological processes, including PD-L1 expression and the PD-1 checkpoint pathway, intracellular transport regulation, cell cycle, and programmed cell death. Notably, the PD-L1 / PD-1 checkpoint pathway was the most significantly enriched pathway, suggesting that DEPs may participate in the pathogenesis of IgG4-RD by regulating immune checkpoint signaling and cell death mechanisms.

[0042] To further elucidate the biological functions of DEPs, GO and KEGG enrichment analyses were performed. GO enrichment analysis revealed that DEPs were significantly enriched in terms related to immune regulation and cellular organization, including "T cell anergy," "T cell anergy regulation," "lymphocyte anergy," and "lymphocyte anergy regulation," highlighting their crucial role in immune exhaustion and tolerance mechanisms closely associated with PD-1 / PD-L1 signaling. Additional enrichment in "macromolecular localization," "RNA transport," and "telomere maintenance" suggests involvement in fundamental cellular processes supporting the observed immunophenotype. Upregulated DEPs were enriched in nuclear / RNA and chromosome organization-related GO terms, suggesting enhanced transcriptional and post-transcriptional regulation, consistent with gene expression reprogramming associated with PD-1 / PD-L1-related T cell exhaustion / tolerance. Conversely, downregulated DEPs were enriched in cell membrane components and cellular import / transport processes, consistent with a weakened membrane-related functional state parallel to immunoreactivity.

[0043] KEGG TopMapStat analysis categorized DEPs into functional groups, with the largest groups being "endocytosis," "PI3K-Akt signaling pathway," "MAPK signaling pathway," and "cancer pathway," indicating complex interactions between cell signaling, immune regulation, and oncogenesis. KEGG pathway enrichment analysis further confirmed these findings, showing significant enrichment in "PD-L1 expression and the PD-1 checkpoint pathway in cancer," directly suggesting the involvement of this axis in the pathophysiology of IgG4-RD. Other significantly enriched pathways included "endocytosis," "necrosis and apoptosis," "oxidative phosphorylation," and "antigen processing and presentation," suggesting that immune checkpoint dysregulation is accompanied by alterations in cell metabolism and antigen presentation.

[0044] Furthermore, GSEA revealed that four hallmark pathways were significantly enriched in IgG4-RD compared to healthy controls. Upregulated mTORC1 and PI3K-AKT-mTOR signaling suggest metabolically highly active B and T cells, potentially driving plasma cell differentiation and pathological antibody production. Significantly enriched oxidative phosphorylation provides evidence supporting enhanced mitochondrial metabolism in support of chronic immune activation. Enrichment of the epithelial-mesenchymal transition (EMT) pathway suggests active tissue remodeling and fibrosis, characteristic of IgG4-RD.

[0045] 2.2 Crosstalk between PD-L1 signaling and immune-metabolic pathways in IgG4-RD To elucidate the regulatory processes governing immune checkpoint signaling, we focused on dendritic enzymes (DEPs) associated with PD-L1 expression and the PD-1 checkpoint pathway. By cross-referencing the DEP list with genes in the KEGG PD-L1 / PD-1 pathway, we identified six key proteins (such as...). Figure 1 Volcano plots showed that JAK2, PTEN, and PTPN6 were significantly upregulated in patients with IgG4-RD, while RPS6KB1, NFATC1, and CD3E were downregulated (e.g., Figure 2 ).

[0046] To understand the interactions between these proteins, a protein-protein interaction (PPI) network of all DEPs was constructed (e.g., Figure 3 Subsequent pivot protein analysis identified JAK2 and PTEN as highly connected regulatory nodes, suggesting their crucial roles in the network (e.g., Figure 4 Subnetwork analysis revealed that PD-L1-related DEPs are integrated into a regulatory network with significance for immune checkpoint signal transduction (e.g., Figure 5PTEN occupies a central position and is linked to HSP90AA1 and SUMO2. Notably, PTEN loss has been shown to increase PD-L1 protein expression via the PI3K / AKT pathway, while HSP90 stabilizes PD-L1 expression, and SUMO2 regulates PD-L1 post-translational modifications. JAK2 is linked to IRF4 and SHC1, which mediate the IFN-gamma / JAK / STAT axis and directly promote PD-L1 transcription. NFATC1 is linked to the cytoskeletal proteins MYH9 and FLNA, which are involved in immune synapse formation and T-cell receptor signaling negatively regulated by the PD-1 / PD-L1 axis. RPS6KB1 interacts with metabolic regulators, reflecting its role in the mTORC1 pathway, which controls PD-L1 expression through translational regulation.

[0047] 2.3 ELISA validation confirmed the dysregulation of PTEN and NFATC1 in IgG4-RD patients. To validate the proteomics findings, ELISA was performed to quantify the levels of PD-L1-related proteins in the plasma of IgG4-RD patients and healthy controls (e.g., ...). Figure 6 Of the six PD-L1-related DEPs, PTEN and NFATC1 were preferentially used for validation because PTEN was identified as a highly connected hub protein in the network, while NFATC1 is a key transcription factor for T cell activation suppressed by PD-1 signaling. Results showed that PTEN was significantly upregulated and NFATC1 was significantly downregulated in IgG4-RD patients compared to healthy controls, consistent with proteomic trends. PTEN is a key negative regulator of the PI3K-Akt pathway; its upregulation further suppresses PI3K / Akt-driven proactivating signaling, thereby enhancing the inhibitory output of the PD-1 / PD-L1 axis on this pathway. NFATC1 acts downstream of the TCR-Ca2+ / calcineurin-NFAT activation cascade; its downregulation indicates a weakened TCR-driven transcriptional activation program, which in turn reduces the ability of T cells to resist checkpoint inhibition, making PD-1 / PD-L1-mediated co-inhibitory signaling more likely to dominate. In summary, these changes contribute to maintaining an immunosuppressive / exhausted state and may lead to the immune dysregulation characteristic of IgG4-RD.

[0048] Mechanistically, six PD-L1-related DEPs regulate the PD-1 / PD-L1 pathway in IgG4-RD. Downregulated RPS6KB1 (S6K) reduces mTORC1-S6K output, weakening T cell activation and making cells more susceptible to PD-1 / PD-L1-mediated inhibition. Downregulated NFATC1 (NFAT), a transcription factor in the TCR-NFAT pathway, impairs the transcriptional synthesis of essential cytokines, reduces T cell activation, and, along with enhanced inhibitory checkpoint signaling, leads to functional T cell exhaustion. Downregulated CD3E, a core component of the TCR complex, further impairs the efficient TCR activation pathway, favoring receptor-mediated inhibition subsequently taken over by the PD-1 pathway. Conversely, upregulated JAK2 (IL-6R-JAK2-STAT3 and IFNGR-JAK2-STAT1 signaling) enhances cytokine receptor signaling, directly leading to upregulated PD-L1 transcription and expression. Upregulated PTEN inhibits PI3K-AKT-mTOR (CD28-associated co-stimulation), thereby reducing activation signaling and favoring PD-1 / PD-L1 checkpoint dominance. Upregulated PTPN6 (SHP-1) is a key phosphatase in the TCR inhibitory cascade, reducing the effectiveness of TCR-mediated signaling, decreasing T cell activation, and being associated with overall T cell effector function. Therefore, SHP-1 provides broad regulation of immune signaling receptor thresholds, enhancing PD-1 / PD-L1-mediated inhibition.

[0049] The coordinated changes—downregulation of RPS6KB1 (S6K), NFATC1 (NFAT), and CD3E impairs T cell activation and effector function, while upregulation of JAK2, PTEN, and PTPN6 (SHP-1) leads to PD-L1 expression and enhanced inhibitory checkpoint signaling—alter the immune homeostasis in IgG4-RD to a state of T cell activation deficiency but increased PD-1 / PD-L1 activity, mechanistically explaining the coexistence of chronic inflammation, immune exhaustion, and progressive fibrosis in this disease.

[0050] 2.4 Validation of PD-L1 modulators as drug treatments for IgG4-RD To explore therapeutics targeting the PD-1 / PD-L1 axis in IgG4-RD, a drug repositioning analysis (DSigDB) was performed, focusing on six major PD-L1-related drug efficacy interventions (DEPs). The top-ranked compounds suggested were resveratrol (a polyphenol natural product) and sorafenib (a multi-kinase inhibitor), showing promise as small molecules. Compound-protein interaction heatmaps revealed that resveratrol is strongly associated with JAK2 and NFATC1, while sorafenib was predicted to interact with and regulate JAK2 and PTEN, suggesting complementary regulatory effects on PD-L1 signaling.

[0051] Mechanistically, resveratrol exerts its immunomodulatory effects by inhibiting the JAK2 / STAT3 and NF-kappaB signaling pathways, leading to downregulation of PD-L1 transcription and partial recovery of T cell function. Sorafenib, on the other hand, indirectly increases checkpoint inhibition by blocking the downstream PI3K-AKT-mTOR signaling pathway following tyrosine kinase activity inhibition, while simultaneously weakening the pro-inflammatory signaling cascade. These mechanisms suggest that resveratrol may alleviate immune exhaustion, while sorafenib controls overactivation, potentially leading to a synergistic effect of immune rebalancing and anti-fibrotic effects.

[0052] In the context of IgG4-RD, the combination of resveratrol and sorafenib may offer dual therapeutic advantages: resveratrol restores T-cell activity and reduces fibrotic signaling, while sorafenib acts on kinase-driven immune activation and fibrosis. Despite limited clinical evidence, their distinct but complementary modes of action suggest a potential combination therapy for long-term disease control.

[0053] The above is a further detailed description of the present invention and should not be considered as a limitation on the specific implementation of the present invention. For those skilled in the art, simple deductions or substitutions without departing from the concept of the present invention are all within the protection scope of the present invention.

Claims

1. The use of a reagent in the preparation of a product for detecting IgG4-RD, characterized in that, The reagent is used to detect the level of PD-1 / PD-L1 axis markers.

2. The application according to claim 1, characterized in that, The PD-1 / PD-L1 axis markers are NFATC1 and PTEN proteins.

3. The application according to claim 1, characterized in that, The PD-1 / PD-L1 axis markers are RPS6KB1, NFATC1, CD3E, PTEN, PTPN6, and JAK2 proteins.

4. The application according to claim 1, characterized in that, The product is selected from reagent kits, protein chips, test strips, or immunoassay devices.

5. The application according to any one of claims 1, 3, or 4, characterized in that, The immune detection device includes: A quantitative system for detecting PD-1 / PD-L1 axis marker levels in quantitative samples; The analysis system uses a quantitative device to detect the levels of PD-1 / PD-L1 axis markers and compares them with normal expression levels. The result output system is used to output the analysis results of the analysis device.

6. The application according to claim 5, characterized in that, The higher-than-normal expression levels of JAK2, PTEN, and PTPN6, and the lower-than-normal expression levels of RPS6KB1, NFATC1, and CD3E, indicate IgG4-RD.

7. The application according to claim 5, characterized in that, The sample is a peripheral blood sample from the subject.

8. The use of a reagent in the preparation of a product for treating IgG4-RD, characterized in that, The reagent is a PD-1 / PD-L1 axis modulator, inhibitor, or agonist.

9. The application according to claim 8, characterized in that, The PD-1 / PD-L1 axis regulator, inhibitor, or agonist is a regulator, inhibitor, or agonist targeting the proteins RPS6KB1, NFATC1, CD3E, PTEN, PTPN6, and JAK2.

10. The application according to claim 8, characterized in that, The PD-1 / PD-L1 axis modulator, inhibitor, or agonist is selected from at least one of resveratrol or sorafenib.

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