Emerging evidence links reactive oxygen species modulator 1 (ROMO1), a key mitochondrial ROS regulator, to rheumatoid arthritis (RA) pathogenesis. However, its exact mechanism remains elusive given the conflicting evidence about its specific function. We used a four-level integrative framework combining multi-omics data and literature‑supported mechanistic inference. At the genetic level, Mendelian randomization (MR) was performed to explore potential causal relationships between ROMO1, IL2RA, HLA-DR, MIF, and RA risk, followed by differential expression analysis and machine learning-based feature selection to identify key mROS genes. The temporal expression dynamics of ROMO1 were assessed in RA progression. At the cellular and tissue levels, we integrated single-cell RNA sequencing and spatial transcriptomics to map cell-type-specific expression and synovial localization of ROMO1-related immune cells and pathways. Finally, our multi-omics findings were contextualized with literature-supported mechanistic inference. (1) MR results were consistent with a potential protective effect of ROMO1 on RA (OR = 0.52) and its potential regulation of risk factors IL2RA (OR = 0.46) and HLA-DR (OR = 0.40). Conversely, IL2RA (OR = 1.42), HLA-DR (OR = 1.88), and MIF (OR = 1.17) were positively associated with RA risk. Additionally, ROMO1 was identified as a top candidate diagnostic predictor with stage-specific dynamics: downregulated in the early but upregulated in the late/remission stages. (2) Single-cell RNA sequencing showed ROMO1's cell-specific expression in CD14+ HLA-DR+ CD74+ monocytes and CD4+ IL2RA+ T cells. Cell communication analysis further suggested that these cells may participate in MIF pathway regulation. Spatial transcriptomics subsequently identified that ROMO1-related cells localized to synovial pathological regions, with MIF pathway changes correlated with RA progression. (3) Finally, literature-supported mechanistic inference suggests that ROMO1 may modulate mROS levels to promote anti-inflammatory M2 macrophage polarization, which could theoretically contribute to reduced systemic inflammation and the alleviation of multi-organ decline in RA. This integrated multi-omics investigation, supported by literature-based mechanistic inference, suggests ROMO1 as a stage-dependent biomarker candidate and potential immune regulator in RA.
Characterized by renal inflammation and structural damage, lupus nephritis (LN) is a severe and often debilitating complication of systemic lupus erythematosus. The dynamic regulation of RNA processing, stability, and translation by N6-methyladenosine (m6A) modification has been implicated in the pathophysiology of LN. As an m6A reader, insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) plays a crucial role in the pathogenesis of various kidney diseases, yet its precise function in LN remains unclear. IGF2BP2 expression was measured in kidney specimens from LN patients and MRL/LPR mice. MRL/lpr mice were administered IGF2BP2 short hairpin RNA adeno-associated viral vectors, followed by evaluation of proteinuria, renal function, and histology. In vitro experiments employed lipopolysaccharide (LPS) stimulation of human kidney tubular epithelial cells (HK-2) to simulate LN inflammatory responses and further investigated the mechanism of IGF2BP2. Molecular interactions were validated via RNA immunoprecipitation and luciferase reporter assays. IGF2BP2 expression is significantly upregulated in LN and positively correlated with proinflammatory factor levels. Knockdown of IGF2BP2 significantly improved renal injury and alleviated renal inflammatory responses in MRL/LPR mice, as well as reduced LPS-induced inflammatory responses in HK-2 cells. Mechanistically, under LN pathological conditions, IGF2BP2 recognized m6A modifications on signal transducer and activator of transcription 1 (STAT1) mRNA and enhanced its stability, leading to the activation of proinflammatory signaling, exacerbation of renal inflammation, and ultimately renal injury. The m6A reader IGF2BP2 promotes proinflammatory signaling in LN by recognizing m6A-modified STAT1 mRNA. Targeting the IGF2BP2-STAT1 axis may represent a potential therapeutic strategy to ameliorate renal inflammatory responses in LN.
Tc17 cells and Tc17.1 cells contribute synovial inflammation during the development of rheumatoid arthritis (RA). However, the modulatory mechanisms of synovial Tc17 cells and Tc17.1 cells remain elusive. In the current study, we performed transcriptome profiling analysis on sorted live synovial Tc17 cells and Tc17.1 cells from a mouse model of collagen-induced arthritis (CIA). We found an array of differentially expressed genes (DEGs), which might be involved in the metabolism, cytokine signaling, and other biological aspects in synovial Tc17 and Tc17.1 cells. Importantly, we identified zinc finger protein 24 (ZNF24), a transcription factor, among the DEGs. ZNF24 knockdown impaired Tc17 differentiation and function in vitro, as evidenced by lower expression of IL-17A, RORγt, GM-CSF, IL-22, perforin, and granzyme B. ZNF24 knockdown resulted in similar effects on Tc17.1 differentiation and function in vitro. Furthermore, ZNF24 knockdown weakened the effects of Tc17 cells and Tc17.1 cells on inducing pro-inflammatory mediator expression in synovial fibroblasts. Adoptive transfer of Tc17.1 cells significantly exacerbated CIA development, whereas ZNF24-knockdown ameliorated the detrimental effect of Tc17.1 cells. Synovial fibroblasts up-regulated ZNF24 expression in Tc17 and Tc17.1 cells in a direct contact manner. The expression pattern and effect of ZNF24 were also observed in patients with RA and human Tc17 cells and Tc17.1 cells. Taken together, our study suggests that ZNF24 is a positive regulatory factor for Tc17 and Tc17.1 cells to exert pro-inflammatory effects in RA. We thus unveiled a novel mechanism by which the functions of synovial Tc17 cells and Tc17.1 cells are modulated.
Diffuse alveolar hemorrhage (DAH) is a lethal, female-biased complication of systemic lupus erythematosus (SLE). While adaptive immunity is a known driver, the role of innate lymphoid cells (ILCs) in DAH and the contribution of sex hormones to this process remain unclear. To characterize pulmonary ILC remodeling during DAH and evaluate whether androgen signaling modulates natural killer (NK) cell homeostasis and disease severity. A pristane-induced DAH model was established in C57BL/6J and Rag2 -/- mice. Pulmonary ILCs were profiled by single-cell transcriptomics and flow cytometry. Baseline sex differences were compared between healthy humans and mice. In vivo interventions included male castration, female testosterone propionate (TP) administration, and antibody-mediated NK cell depletion. DAH was accompanied by a marked reduction in pulmonary NK cell frequency in both C57BL/6J and Rag2 -/- mice, indicating an innate feature independent of adaptive lymphocytes. DAH-associated NK cells displayed an activated but constrained state, showing increased expression of effector-associated genes Prf1 and Ifng, together with the inhibitory receptor gene Lag3. At baseline, healthy human females and female mice exhibited lower NK cell frequencies than males. While male castration did not significantly alter DAH severity, TP administration in females restored pulmonary NK cell dominance and significantly attenuated disease severity. NK cell depletion prior to challenge led to a trend toward more severe hemorrhage (p = 0.084). Pulmonary NK cells are reduced in a lupus-associated DAH model and may participate in disease development. Androgen treatment restores pulmonary NK cell predominance and attenuates DAH severity, supporting further investigation of strategies aimed at restoring or preserving pulmonary NK cell homeostasis and function as potential therapeutic approaches for DAH.
Interleukin 2 (IL-2) is a cytokine essential for the development and function of regulatory T cells, which are key players in maintaining immune tolerance by suppressing autoreactive lymphocytes. Patients with systemic lupus erythematosus (SLE) often exhibit reduced IL-2 production, resulting in impaired Treg function and a consequent breakdown of immune tolerance. Hematological abnormalities, mainly leukopenia and lymphopenia, are prominent features of SLE. This study aimed to explore the relationship between serum IL-2 concentrations and the altered blood cell populations observed in patients with SLE. We recruited 235 patients with SLE. Complete blood counts, including red blood cells, white blood cells, and platelets, were obtained, and IL-2 levels were quantified using the ultrasensitive single-molecule array (Simoa) technique. Multivariate linear and logistic regression analyses were performed to evaluate the associations between IL-2 concentrations and hematological parameters. Multivariate linear regression revealed significant inverse associations between the serum IL-2 concentration and the hemoglobin level, hematocrit, mean corpuscular hemoglobin concentration, and total leukocyte, neutrophil, lymphocyte, and basophil counts. Similarly, after adjustment for covariates, serum IL-2 was associated with increased odds of anemia, leukopenia, and lymphopenia but not neutropenia or thrombocytopenia. In conclusion, circulating IL-2 levels are independently associated with specific hematological abnormalities, including anemia, leukopenia, and lymphopenia, in patients with SLE. These findings highlight the potential role of IL-2 and regulatory T cell dysfunction in the hematologic manifestations of SLE.
The advancement of hepatocellular carcinoma (HCC) is associated with metabolic reprogramming and immune escape. Compound kushen injection (CKI) is a cancer therapeutic agent with a long history. O-GlcNAcylation is reported to regulate tumor progression. We aimed to investigate how CKI affects HCC cell glycolysis and immune evasion and then clarify the underlying O-GlcNAcylation regulatory mechanism. HCC cell lines and subcutaneous transplantation mice were treated with CKI, and glycolysis and immune evasion were analyzed. The O-GlcNAcylation was analyzed using immunoprecipitation and western blotting. The results showed that CKI inhibited the viability, glycolysis, and immune escape of HCC cells, as well as hindered tumor growth, glycolysis, and immune evasion in vivo. Induction of glycolysis caused by hypoxia reduced the effect of CKI on immune evasion. Besides, CKI reduced the O-GlcNAc levels and OGT transcription activity. Overexpression of OGT counteracted the inhibition of immune evasion caused by CKI. Additionally, OGT catalyzed the O-GlcNAcylation of PD-L1 and enhanced its stability. In conclusion, CKI suppresses immune evasion of HCC cells in association with glycolysis suppression, which is related to the reduction of PGT-mediated PD-L1 O-GlcNAcylation. These findings suggest that CKI possesses an anti-HCC effect, and targeting PD-L1 O-GlcNAcylation may improve its therapeutic efficacy on HCC.
Increasing evidence shows that CD8+ T cells are the pathogenic mediators of tissue injury in systemic lupus erythematosus (SLE), sustaining the chronic inflammation through the accumulation of long-lived cytotoxic memory populations. However, the transcriptional mechanisms that prevent the aberrant differentiation of pathogenic CD8+ T cells remain poorly understood. Here, the transcription factor E4BP4 (NFIL3) was identified as a critical restraint of cytotoxic effector-memory CD8+ T cells in lupus. E4BP4 expression was reduced in CD8+ T cells from SLE patients and inversely correlated with disease activity. Using a lupus-like disease model, we found that E4BP4 deficiency accelerated disease progression, resulting in heightened autoantibody production, immune complex deposition, and renal pathology. This phenotype was associated with the systemic accumulation of cytotoxic effector-memory CD8+ T cells. Depletion of CD8+ T cells significantly ameliorated the disease phenotype, confirming the functional contribution of CD8+ T cells to lupus-like immunopathology. Competitive adoptive transfer experiments revealed that E4BP4 functions cell-intrinsically to limit the cytotoxicity and proliferation of CD8+ T cells in autoimmunity. Beyond autoimmunity, E4BP4 deficiency also resulted in an exuberant CD8+ effector-memory T cell response to Listeria monocytogenes infection, indicating a broader role for E4BP4 in limiting CD8+ T cell effector-memory responses. Collectively, these findings establish E4BP4 as a transcriptional checkpoint that restricts pathogenic CD8+ effector-memory T cell responses to maintain immune homeostasis in autoimmunity and infection.
Myasthenia gravis (MG) is a complex autoimmune neuromuscular disorder, and the role of lipid metabolism dysregulation in MG pathogenesis remains unclear. This study aimed to investigate the molecular mechanisms of lipid metabolism regulation in myasthenia gravis development by integrating single-cell and bulk transcriptomic data. This study analyzed bulk RNA sequencing data from the GSE85452 dataset (13 MG patients and 12 healthy controls) and single-cell RNA sequencing data from the GSE227835 dataset (10 MG patients and 10 healthy controls). Differential expression analysis was performed, and weighted gene co-expression network analysis (WGCNA) was conducted to identify disease-related modules. Key genes were screened through the intersection of differentially expressed genes (DEGs), WGCNA hub genes, and lipid metabolism-related genes. Functional enrichment analysis, protein‒protein interaction (PPI) network construction, immune infiltration analysis, and regulatory network analysis were performed. Single-cell analysis was used to characterize cellular heterogeneity and intercellular communication features. A nomogram prediction model was constructed and internally validated using leave-one-out cross-validation (LOOCV). Potential therapeutic compounds were identified through drug prediction and molecular docking analysis. Furthermore, key genes were validated by RT-qPCR in an independent cohort of 5 MG patients and 5 healthy controls. A total of 823 DEGs and 13 co-expression modules were identified, of which 4 modules were significantly associated with MG. Twenty-one candidate genes were screened, and 2 key genes (IRS2 and ALDH2) were ultimately determined. IRS2 was significantly downregulated while ALDH2 was significantly upregulated in MG patients. The nomogram model based on key genes demonstrated excellent predictive performance (AUC = 0.897). Immune infiltration analysis showed increased regulatory T cells (Tregs) and decreased CD4+ memory activated T cells in MG patients. Single-cell analysis identified 10 major cell types. Cell‒cell communication analysis revealed dense interactions among CD4+ T cells, B cells, and CD14+ monocytes. Drug prediction identified metformin and cyclophosphamide as potential therapeutic candidates, and molecular docking confirmed favorable binding affinities. This integrative study generated the hypothesis that lipid metabolism dysregulation, potentially mediated by IRS2 and ALDH2, may contribute to immune dysfunction in MG pathogenesis. These findings provide preliminary insights into the molecular mechanisms underlying MG development and suggest potential diagnostic biomarkers and therapeutic targets. However, these results are hypothesis-generating, and clinical translation is contingent upon rigorous mechanistic validation through functional experiments, animal models, and larger multicenter clinical cohorts.
While interferon regulatory factor 1 (IRF1) has been implicated in reactive oxygen species (ROS)-dependent neutrophil extracellular trap (NET) release during acute lung injury, its regulatory role in mitochondrial dysfunction-driven NETosis specific to pediatric pneumonia remains unexplored. Neutrophils were isolated from the bronchoalveolar lavage fluid (BALF) of pediatric pneumonia patients and the bone marrow of mice. IRF1 expression was quantified via qRT-PCR and Western blotting. Mitochondrial ROS (mtROS) and total ROS were measured by flow cytometry. Mitochondrial dysfunction was assessed by ATP quantification. NETosis was evaluated through immunofluorescence staining and ELISA quantification of neutrophil elastase (NE), myeloperoxidase-DNA (MPO-DNA), and citrullinated histone H3 (Cit-H3) levels. For in vivo studies, pneumonia-related lung injury was induced by intratracheal LPS instillation in mice, with pathological severity graded by H&E staining, pulmonary edema quantified via the wet/dry weight ratio, and inflammation assessed by the BALF protein concentration. Mechanistically, the IRF1/MMP9 interaction was predicted by bioinformatics (STRING database) and validated by co-immunoprecipitation (Co-IP) and immunofluorescence staining, while MMP9 overexpression was achieved via lentivirus transduction to delineate pathway regulation. IRF1 was significantly upregulated in BALF neutrophils from patients and correlated with elevated ROS production and mitochondrial dysfunction, as well as NETs release. IRF1 knockdown attenuated ROS-driven NETosis in vitro. Matrix metalloproteinase 9 (MMP9) was predicted to interact with IRF1, and MMP9 overexpression effectively reversed the beneficial effects of IRF1 deficiency on ROS release, mitochondrial dysfunction, neutrophil apoptosis, and NETosis. Consistently, in mouse models, MMP9 overexpression abolished the protective effects of IRF1 deficiency, exacerbating acute lung injury and restoring NETs levels in BALF.
OBJECTIVE:To investigate the effects of Platycodin D (PD) on the proliferation, migration, and angiogenesis of EA.hy926 endothelial cells stimulated by supernatant from a rheumatoid arthritis (RA) synovial cell model (MH7A), this study aims to preliminarily explore its potential mechanisms. METHODS:The TNF-α-stimulated MH7A cells were utilized as a model for rheumatoid arthritis (RA) cells. The supernatant from these cells was collected and designated as conditioned medium (CM), which was then used to stimulate EA.hy926 cells, thereby establishing an RA endothelial cell model; EA.hy926 cells were transduced with lentivirus to overexpress or knock down CD146; The experimental groups included: a normal control group, a model group (optimal CM stimulation group), and PD treatment groups (PD at concentrations of 1.25, 2.5, and 5 mg/L); sh-CD146 + CM, sh-NC + CM, sh-NC, lenti-CD146 + CM, lenti-CD146-NC. The rescue experiment included the following groups: CM, CM + PD group, CM + lenti-CD146 group, and CM + lenti-CD146 + PD treatment group. Cell proliferation was assessed using the CCK-8 assay; cell migration was evaluated through the wound-healing assay; angiogenic capability was determined via tube formation assays; and CD146 expression levels were measured by Western blotting. RESULTS:Compared to the normal group, CM stimulation significantly enhanced endothelial cell proliferation (P < 0.01). However, after 24 hours of PD treatment, a notable decrease in cell proliferation was observed (P < 0.05). Additionally, CM stimulation improved cell migration ability relative to the normal group; this enhancement was significantly diminished following PD treatment (P < 0.01). Furthermore, tube formation capability was markedly increased with CM stimulation compared to the normal group, but showed significant inhibition after 24 hours of PD treatment (P < 0.0001). Moreover, CD146 expression levels were significantly elevated in the model group when compared to the normal group and subsequently decreased following PD treatment (P < 0.01), and in a dose-dependent manner. In the rescue experiment, compared with the PD treatment group (CM + PD 5 mg/L), the proliferation, migration, and tube formation abilities of cells in the CD146 overexpression + PD treatment group (CM + lenti-CD146 + PD 5 mg/L) were significantly increased (P < 0.0001, P < 0.05, P < 0.001). Interestingly, when compared with the overexpression group (CM + lenti-CD146), these abilities in the overexpression+ PD treatment group (CM + lenti-CD146 + PD 5 mg/L) remained significantly suppressed (P < 0.0001, P < 0.0001, P < 0.05). CONCLUSION:In this study, we further found that CD146 exhibits significant pro-angiogenic, pro-migratory, and pro-proliferative effects in EA.hy926 cells. Meanwhile, PD was shown to significantly inhibit the angiogenesis, proliferation, and migration of EA.hy926 cells in a dose-dependent manner, and downregulate CD146 expression in a dose-dependent manner. CD146 overexpression partially attenuated PD's anti-angiogenic effect, but PD still exhibited strong anti-angiogenic activity in CD146-overexpressing cells, suggesting that PD acts partly through CD146 regulation and possibly other pathways, reflecting its multifaceted nature. This study is the first to link PD's anti-angiogenic effect to CD146, revealing a promising therapeutic direction for RA.
Hashimoto's thyroiditis (HT) is a common autoimmune thyroid disorder with a complex genetic background. Interleukin-6 (IL-6) has been implicated in its pathogenesis, but the genetic mechanisms remain unclear. Two-sample Mendelian randomization (MR) using expression quantitative trait loci (eQTL) and gene set enrichment analysis (GWAS) data identified IL‑6-HT genes. In vivo validation in nonobese diabetic (NOD) mice involved siRNA knockdown, histopathology HE/Immunohistochemistry (HE/IHC), and enzyme-linked immunosorbent assay (ELISA). GSEA and molecular docking explored mechanisms. Western blot in THP‑1 cells assessed Nuclear factor kappa-light-chain-enhancer of activated B cells (NF‑κB) pathway proteins (pIKKβ, IKKβ, pTAK1, TAK1, pp65, p65) and associations with SLC45A3/CCDC77 at 2, 24, 48 h (β‑actin control). MR analysis identified CCDC77 and SLC45A3 as key genes associated with both IL-6 and HT. In vivo experiments showed that knockdown of CCDC77 exacerbated thyroid damage, increased serum anti-thyroperoxidase antibody (TPOAb), anti-thyroglobulin antibody (TGAb), and IL-6 levels, while reduced free triiodothyronine (FT3) and free thyroxine (FT4). Conversely, SLC45A3 knockdown alleviated thyroiditis, reduced antibody levels, and improved thyroid function. GSEA Western blot analysis further revealed that both CCDC77 and SLC45A3 were involved in regulating the activation of NF‑κB signaling pathway. Our study demonstrated that SLC45A3 and CCDC77 were involved in modulating HT progression through NF‑κB-IL-6 axis, providing novel therapeutic strategies for clinical applications.
Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease with complex molecular mechanisms. Although transcriptomic studies have revealed prominent interferon signatures in SLE, robust prioritization of candidate genes across independent cohorts remains challenging. In this study, we performed a multi-cohort transcriptomic analysis using publicly available GEO datasets. Differential expression analysis was conducted independently within each cohort to minimize cross-study confounding. Machine learning models, including LASSO, support vector machine, and random forest, were applied for feature prioritization in a designated training cohort, followed by independent validation in separate datasets. Model interpretability was assessed using SHAP analysis. Immune cell composition was estimated descriptively using CIBERSORT. In addition, molecular docking and molecular dynamics simulations were performed as exploratory in silico analyses to evaluate potential protein-compound interactions. A set of consistently dysregulated genes across cohorts was identified, many of which are associated with interferon signaling. Among these, RSAD2 showed robust prioritization across multiple machine learning models. The predictive performance of the models was stable in independent validation datasets. SHAP analysis highlighted the contribution of interferon-stimulated genes to model predictions. Immune deconvolution suggested altered immune cell composition in SLE samples, consistent with previously reported immune activation patterns. Exploratory in silico analyses suggested a potential interaction between artemisinin and RSAD2. This study provides a robust, multi-cohort computational framework for prioritizing candidate genes associated with SLE. The findings highlight interferon-associated transcriptional features as reproducible molecular signatures of SLE and generate testable hypotheses for future experimental and clinical investigation.
IgA vasculitis (IgAV) primarily affects small vessels, but rare cases with necrotizing arteritis (NA) raise questions about overlap with polyarteritis nodosa (PAN). To characterize IgAV with necrotizing arteritis (IgAV-NA) and compare its phenotype with classical IgAV and PAN. We performed a multicenter retrospective study combined with a systematic literature review (1990-2025). Patients fulfilled EULAR/PRINTO/PRES IgAV criteria, had pathological or imaging evidence of NA in small or medium arteries, and were ANCA-negative. Thirty patients were included (7 from databases, 23 from the literature). NA was confirmed by biopsy (n = 16) or vascular imaging (n = 14). Clinical features, treatments, remission, and mortality were compared with 257 adult IgAV and 196 PAN patients. Median age was 54.5 years. IgAV-NA was characterized by severe manifestations, including gastrointestinal bleeding, perforation, surgical abdomen, neuropathy, pancreatitis, and livedo. Compared with classical IgAV, IgAV-NA showed significantly higher rates of multi-organ involvement and mortality. Compared with PAN, IgAV-NA shared vascular complications but had less fever and neuropathy. Despite arterial involvement, patients did not fulfil PAN criteria. IgAV-NA represents a rare, severe IgAV phenotype with life-threatening complications rather than an IgAV-PAN overlap. Severe or atypical IgAV presentations should prompt vascular imaging and intensified immunosuppression.
To systematically examine global research trends in the association between ferroptosis and autoimmune diseases from 2018 to 2025, thereby informing future mechanistic and translational investigations in this field. On March 17, 2026, relevant publications were obtained from the Web of Science Core Collection and PubMed databases. This included articles and reviews written in English and published between 2018 and 2025. Bibliometric visualization was conducted using CiteSpace 6.4.R1, VOSviewer 1.6.20, and Scimago Graphica to map countries, authors, institutions, keywords, journals, and references. Data processing and descriptive statistical analysis were performed using Microsoft Excel 2019. Seven hundred and nineteen publications from 27 countries were included in the analysis. The annual number of publications showed a consistent upward trend, with China contributing the largest share (487 publications). High-frequency keywords included rheumatoid arthritis, ulcerative colitis, inflammatory bowel disease, oxidative stress, and lipid peroxidation, indicating research hotspots in this field. Research on the association between ferroptosis and autoimmune diseases has expanded substantially over the past 8 years. However, the field remains in a relatively early stage of development. These findings suggest that the field is receiving increasing scholarly attention and may provide useful references for future biomarker discovery and novel therapeutic strategy development.
Monocytes are major contributors to systemic lupus erythematosus (SLE) pathogenesis, modulating B- and T-cell autoreactivity through inflammatory cytokine secretion and disrupted immune clearance. Under prolonged inflammatory pressure, monocytes develop a pathogenic “exhausted” immune memory state defined by paradoxical proinflammatory and immunosuppressive gene expression and diminished immune effector functions. To determine whether chronic inflammation in SLE elicits a similar monocyte exhaustion phenotype, we analyzed bone marrow and splenic reservoir monocytes from lupus-prone MRL and MRL/lpr (LPR) mice. Monocytes from LPR mice exhibit chronic immune exhaustion, including reduced chemokine receptor CX3CR1 cell surface levels, heightened sensitivity to endotoxin stress, and altered expression of T cell regulatory molecules PD-L1, CD200R, ICOS-L, and CD86. Exhaustion severity correlated with SLE progression, although the development of monocyte exhaustion preceded the onset of disease symptoms. These features were largely recapitulated in monocytes from patients with SLE, demonstrating that monocyte exhaustion is a generalizable feature of SLE pathogenesis.
Maternal factors during fetal development may influence the onset and progression of autoimmune diseases such as type 1 diabetes (T1D). Using the non-obese diabetic (NOD) mouse model, we investigated whether the in utero environment affects diabetes incidence, pancreatic insulitis, autoantibody profiles, and lymphocyte composition. NOD embryos were transferred to C57BL/6 (B6) females, generating NOD/B6 offspring, and reciprocal transfers produced B6/NOD mice. Diabetes incidence was monitored for 52 weeks, and immune parameters were analyzed at 7-11 weeks of age using immunofluorescence, ELISA, and flow cytometry. NOD/B6 mice exhibited a ~50% reduction in cumulative diabetes incidence compared to NOD controls, despite similar insulitis severity. Serum analysis revealed significantly increased IgM titers against insulin, DNA, and GAD65 in NOD/B6 mice relative to NOD mice, along with altered IgG subclass distributions, including elevated IgG1 and IgG2c levels. UMAP and ROC analyses confirmed that distinct IgG subclass patterns were influenced by the maternal environment. Furthermore, NOD/B6 mice displayed an intermediate phenotype in B cell subset distribution and a notable increase in CD4⁺CD25⁺ regulatory T cells (Tregs) in the blood and spleen, with restored CD25 expression compared to NOD mice. Marginal zone B cells as well as thymic B cell proportions were reduced in NOD/B6 relative to NOD, whereas bone marrow B cell development remained unaffected. These findings demonstrate that maternal factors imprint long-term changes in adaptive immunity and modulate diabetes susceptibility in NOD mice. The observed alterations in antibody profiles and the Treg population suggest that transplacental influences contributed to immune regulation beyond genetic predisposition, highlighting a potential non-genetic contributing factor in T1D pathogenesis.
Systemic lupus erythematosus (SLE) represents a prototypical autoimmune disorder characterized by multi-organ involvement due to loss of immune tolerance. Emerging evidence implicates macrophage polarization dynamics as a pivotal contributor to SLE pathogenesis, with epigenetic regulation by long noncoding RNAs (lncRNAs) emerging as a critical regulatory mechanism. To elucidate the pathogenic role of exosome-derived lncRNAs in this process, we performed comprehensive RNA sequencing analysis on circulating exosomes from SLE patients, identifying lncRNA H19 as a significantly upregulated candidate. Subsequent functional analyses using primary murine macrophages demonstrated that exogenous H19 administration recapitulated the pro-inflammatory phenotype observed in SLE, as evidenced by enhanced M1 macrophage differentiation, transmigration capacity, and renal infiltration patterns in experimental models. Notably, we primarily identified lncRNA H19 as a potential indicator in SLE, the knockdown of which might significantly attenuate these pathological manifestations. Mechanistic investigations revealed that H19 promoted the proliferation and migration of macrophages in vitro and exerted its regulatory function through competitive binding to miR-145-5p, thereby derepressing PAI-1 expression and subsequently activating the JAK2-STAT3 signaling cascade to orchestrate pro-inflammatory cytokine production. Overall, this study reveals novel molecular mechanisms underlying immune dysregulation in SLE, establishing exosomal lncRNA H19 as a key epigenetic modulator of macrophage polarization via the miR-145-5p/PAI-1/STAT3 axis, and proposes a novel potential therapeutic target for restoring immune homeostasis in patients with SLE before clinical practice.
Lupus nephritis (LN) represents the most severe renal manifestation of systemic lupus erythematosus (SLE), contributing to significant morbidity. While current assessments focus on glomerular pathology, tubulointerstitial lesions may offer critical insights into disease progression and treatment response. This study develops a clinical prediction model integrating tubulointerstitial molecular signatures. We performed bioinformatics analysis using two independent tubulointerstitial gene expression datasets (GSE113342 and GSE200306), applying batch effect correction and principal component analysis (PCA) to identify differentially expressed genes (DEGs). A protein‒protein interaction (PPI) network isolated hub genes, and least absolute shrinkage and selection operator (LASSO) regression defined the novel “Nscore” parameter predictive of treatment response. The Nscore, incorporating seven key genes (EGR1, IL6R, TFRC, CCL19, IFI16, IFI35, and Fra1), showed a significant positive correlation with 24-h proteinuria and effectively distinguished complete-response (CR)/partial-response (PR) from non-response (NR). Immune deconvolution using the CIBERSORT algorithm revealed an increased abundance of T follicular helper (Tfh) cells and M1 macrophages in NR samples. A clinical nomogram integrating Nscore and sex demonstrated excellent discrimination. This model combines molecular biomarkers with clinical parameters to improve personalized therapeutic stratification, advancing treatment strategies beyond traditional glomerulocentric paradigms and identifying immune cell signatures as potential targets for immunomodulatory interventions.
Excessive M1 polarization of macrophages is a key driver of inflammatory processes in gout, while the regulatory mechanisms involved remain unclear. Herein, the role of carbohydrate antigen 72-4 (CA72-4), derived from human synovial cells (HFLS), in regulating macrophage M1 polarization during gout progression was investigated. PMA was used to induce differentiation of THP-1 cells into macrophages, and the macrophages were then treated with a conventional medium (CM), the CM was from Non-treated HFLS (NCM), or the CM from MSU-treated HFLS (MCM), and then incubated with MSU treatment for 24 h. Cell viability was assessed by CCK-8 assay. IL-1β, TGF-β1, and CA72-4 levels in the supernatant of macrophages were detected by ELISA. The interaction between CA72-4 and sialic acid-binding immunoglobulin-like lectin-15 (Siglec-15) was analyzed by Co-IP assay. MSU-induced HFLS to secrete CA72-4, and CA72-4 derived from HFLS inhibited MSU-induced macrophage M1 polarization. Mechanistically, CA72-4 activated the TIGIT/SHP-1 signaling in macrophages by binding to Siglec-15 on macrophages. As expected, Siglec-15 knockdown weakened the activation effect of MCM on TIGIT/SHP-1 signaling and the inhibitory effect on macrophage M1 polarization. MSU stimulated HFLS to secrete CA72-4, and CA72-4 derived from HFLS suppressed MSU-induced macrophage M1 polarization by activating TIGIT/SHP-1 signaling through targeting Siglec-15.
Asthma represents a classic respiratory disorder marked by chronic inflammation in the airways. This study aims to demonstrate the mechanism of bronchial epithelial cell-derived extracellular vesicles (BEC-EVs) carrying miR-27b-3p in airway inflammation in asthmatic mice, providing potential therapeutic targets for asthma. BECs and their EVs were isolated from mice and characterized. Asthmatic mouse models were established using ovalbumin and neutrophils were isolated. Histopathological changes and collagen deposition in lung tissues were observed. The levels of proinflammatory factors in the BALF and neutrophils were measured by ELISA. dsDNA levels in BALF or neutrophils were quantified using a dsDNA kit. Expression of cy3-labeled miR-27b-3p in neutrophils was detected. The levels of miR-27b-3p, ZMAT3, FGF1, Cit-H3, and MPO in lung tissues and cells were detected by RT-qPCR and Western blot. The binding relationships between miR-27b-3p and ZMAT3 and between ZMAT3 and FGF1 were verified. Combined experiments were used to validate the molecular mechanism by which BEC-EVs promote NET formation and regulate airway inflammation in asthmatic mice via the miR-27b-3p/ZMAT3/FGF1 axis. After BEC-EVs treatment, peribronchial inflammatory cell infiltration and collagen deposition were aggravated, and Cit-H3, MPO, and dsDNA were increased in OVA-induced mice, indicating exacerbated airway inflammation and promoted NET formation. BEC-EVs delivered miR-27b-3p to neutrophils and upregulated miR-27b-3p expression, which inhibited ZMAT3 to promote FGF1 expression. ZMAT3 overexpression or FGF1 knockdown partially reversed the BEC-EVs-induced NET formation. BEC-EVs promote NET formation and further aggravate airway inflammation in OVA-induced asthmatic mice by delivering miR-27b-3p to neutrophils, inhibiting ZMAT3, and promoting FGF1 expression.
