Standardizing the interpretation of ANA patterns is a persistent challenge in rheumatology due to inter-observer discordance. This research introduces an ensemble deep learning framework optimized for the International Consensus on ANA Patterns (ICAP) system. By integrating ResNet-50 and EfficientNet-B0 via validation-tuned weighting and logit-level averaging, we processed both a single-label benchmark and a high-variability clinical dataset containing overlapping (multi-label) patterns. Our model demonstrated high fidelity in the single-label setting (92.5
Background: Chronic obstructive pulmonary disease (COPD) is a disease characterized by airflow obstruction. Peroxiredoxin 6 (PRDX6) is vital in antioxidant defense and phospholipid metabolism, and linked to disease occurrence and progression. Ubiquitin-specific protease 19 (USP19), a deubiquitinating enzyme involved in protein stability regulation, has been implicated in inflammatory diseases and cellular stress responses, yet its role in COPD remains largely unknown. The objective of this study is to investigate the impact and the underlying mechanism of PRDX6 on cigarette smoke extract (CSE)-induced cell injury and ferroptosis, with a focus on the regulatory roles of USP19 and histone deacetylase 6 (HDAC6).Methods: The expression levels of the genes were detected by reverse transcription-quantitative polymerase chain reaction (qRT-PCR) and western blot. The Cell counting kit-8 (CCK-8) was utilized to assess cell viability. Cell apoptosis was detected by flow cytometry. The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected using the enzyme-linked immunosorbent assay (ELISA). The iron ion (Fe2+), reactive oxygen species (ROS), and glutathione (GSH) kits were used to evaluate the ferroptosis index. The Biological General Repository for Interaction Datasets (BioGRID), Hitpredict database, and Ubibrowser database were used to predict protein interactions. Protein modifications and protein-protein interactions were verified using co-immunoprecipitation (CoIP). The treatment with cycloheximide (CHX) was used to verify protein stability. The rat COPD model was used for in vivo validation. Hematoxylin-eosin (HE), Masson’s trichrome (Masson) and Periodic Acid-Schiff (PAS) staining were utilized for the assessment of lung injury. Immunohistochemistry (IHC) analysis was conducted to assess the levels of PRDX6 present in rat lung tissue.Results: In CSE-induced 16HBE cells, upregulating PRDX6 expression mitigated the decline in cellular viability, prevented apoptosis, suppressed inflammatory reactions, and halted ferroptosis. HDAC6 facilitated CSE-induced cellular damage and ferroptosis by mediating the deacetylation modification of PRDX6. USP19 enhanced the stability of HDAC6 by deubiquitination, thereby exacerbating CSE-induced cellular damage and ferroptosis. The elevated levels of PRDX6 mitigated cellular damage and ferroptosis in the rat COPD models.Conclusions: USP19 modulates PRDX6 expression by deubiquitinating HDAC6, thereby promoting CSE-induced cellular damage and ferroptosis. This deepens our understanding on COPD pathogenic mechanism and may offer a new perspective for COPD treatment.
Chimeric antigen receptors (CARs)-T cell therapy is emerging as a potent approach for autoimmune diseases. However, its application in autoimmune conditions remains limited, and safety outcomes observed in malignancies can’t reliably serve as a reference. Therefore, it’s necessary to summarize the safety profiles in autoimmune diseases to provide evidence for future expanding trials. A systematic review was conducted to analyze the CAR-T therapy safety in rheumatic diseases via database searches up to December 2025. Studies reporting safety data were included, while abstracts, reviews, and cases with malignancies were excluded. Factors associated with cytokine release syndrome (CRS) were analyzed using Firth's penalized logistic regression. This study included 38 studies, involving a total of 115 patients with autoimmune disease. Severe adverse events were rare. CRS and immune effector cell-associated neurotoxicity syndrome (ICANS) occurred in 70.4
Langerhans cells (LCs) are primary immune sentinels at barrier surfaces, acting as the first line of defence against foreign pathogens. However, their low abundance within the human epidermis severely limits their availability for functional and mechanistic studies. To address this challenge, we sought to provide a versatile experimental framework for generating, identifying, and characterizing distinct subsets of LCs derived from in vitro differentiation of human peripheral monocytes. LCs’ heterogeneity has long been considered, with recent studies classifying them into four subsets: effector LCs (LC1) and regulatory LCs (LC2) present at steady state, along with activated LCs (aLC) and migratory LCs (migLC) found during skin inflammation. In this study, LC1, LC2, and aLC/migLC subsets were identified by flow cytometric profiling of CD207, CD1a, CD83, and CD197. LC phenotype was further confirmed by examining HLA-DR, CD14, CD209, CD1b, and CD1c expression. Additionally, the phagocytic capacity of LCs relative to distinct macrophage polarizations was evaluated. From the in vitro-generated LCs, the subsets LC1, LC2, and a joint aLC/migLC cluster were identified. LC1 constituted the predominant population, appearing at higher proportions than both LC2 and the aLC/migLC cluster. The proportions of LC2 and aLC/migLC remained relatively consistent across donors, whereas LC1 abundance showed greater variation, suggesting a higher donor-dependent heterogeneity. Overall, LCs exhibited the lowest phagocytic capacity compared with the different macrophage polarizations. Our study defines LC subsets that can be generated from peripheral monocytes in vitro and provides a framework for deeper exploration of human LC biology, subset heterogeneity, and function in both normal and pathological skin.
TBK1 is a serine-tyrosine kinase protein that transmits signals from pattern recognition receptors to the NF-κB pathway leading to production of Type 1 Interferons. Mutations in this protein have been associated with arthritis, vasculitis, herpes simplex encephalitis and amyotrophic lateral sclerosis. In the current study, we characterized the functional consequences of a TBK1-variant bearing a truncation in exon 4 and 5 in a patient with poly arthritis resembling juvenile idiopathic arthritis and necrotizing encephalitis. The truncation was associated with reduced TBK1 protein abundance and altered phosphorylation. The variant was associated with increased basal/and or Poly-I: C induced IL-6, TNFα, IL-1β and IL-18 and type 1 Interferon ex vivo. Our findings expand the phenotypic spectrum of TBK1 loss-of-function variants and may provide insight into the management of immune dysregulation in affected patients.
Ulcerative colitis (UC) is associated with gut microbial dysbiosis, but the host molecular alterations potentially linked to microbially derived metabolites remain incompletely understood. We integrated Mendelian randomization (MR), microbial metabolite annotation, computational target prediction, colonic transcriptomics, network analysis, and machine learning. MiBioGen microbiome GWAS data were used as exposures and FinnGen Release 12 ULCERENTER as the outcome. Metabolites linked to MR-prioritized taxa were retrieved from GutMGene, and human targets were predicted using SwissTargetPrediction and SEA. UC-related genes were defined by integrating differential expression analysis and WGCNA and then intersected with predicted metabolite targets. MR prioritized one family and eight genera showing nominal genetically supported associations with UC, but none remained significant after Benjamini–Hochberg FDR correction. Three prioritized genera were linked to 15 microbe–metabolite records, corresponding to 13 unique metabolites; nine were retained for target prediction, yielding 277 unique predicted human targets. Transcriptomic analysis identified 1,530 DEGs and a 312-gene MEgrey60 module, with 273 overlapping genes, producing 1,569 unique UC-related genes. Their intersection with the 277 predicted targets yielded 47 candidate genes. Enrichment analyses highlighted mainly metabolic and lipid-related processes. Random Forest showed the highest mean AUC across the two independent external benchmarking cohorts, and SHAP prioritized EPHX1, HSD17B2, IGFBP5, and MMP10. IBDome analysis showed inflammation-associated expression differences in these genes. This study provides a genomics-informed, hypothesis-generating framework that prioritizes candidate microbe–metabolite–host relationships in UC for future experimental validation.
This study aimed to investigate the role of the homeobox gene SIX1 (sine oculis homeobox homolog 1) in acute myeloid leukemia (AML) progression and to explore its underlying metabolic and immunological mechanisms. SIX1 expression was examined in human bone marrow stromal cells (HS-5) and AML cell lines (HL-60, SKM-1, THP-1, KG-1, NB4, and MOLM13) using RT-qPCR. SIX1-knockdown and SIX1-overexpressing MOLM13 cells were generated by RNA interference and plasmid transfection, respectively. Cell cycle distribution, proliferation, and metabolic parameters, including intracellular glucose, lactate, and ATP levels, were assessed using flow cytometry, EdU incorporation, MTT assays, and ELISA. Mitochondrial content was analyzed by transmission electron microscopy. Peripheral blood mononuclear cells (PBMCs) from healthy donors were co-cultured with genetically modified MOLM13 cells under low-glucose conditions to evaluate T-cell proportions and cytokine production (IL-6, TNF-α, and IFN-γ). An AML mouse model was further established to assess the effects of SIX1 on T-cell composition and function by flow cytometry, while global metabolomics was applied to characterize metabolic reprogramming in vivo. SIX1 was significantly up-regulated in AML cells and promoted MOLM13 cell proliferation by enhancing glycolytic metabolism. Moreover, SIX1 suppressed CD8⁺ T-cell effector function both in vitro and in vivo and was associated with extensive alterations in amino acid, glucose, and lipid metabolism in AML mice. SIX1 facilitates AML progression by promoting glycolytic reprogramming in leukemia cells and potentially impairing CD8⁺ T-cell effector function through metabolic competition, highlighting SIX1 as a promising therapeutic target.
Recurrent pregnancy loss (RPL) is clinically defined as two or more pregnancy failures before 20–24 weeks of gestation. Over 50
FLNA encodes filamin A, a ubiquitously expressed actin-binding cytoskeletal protein that cross-links actin filaments and links them to membrane-associated signaling complexes. Although FLNA has been implicated in T-cell signaling and regulatory T-cell development in murine models, its role in human immune-cell function remains incompletely understood. Here, we investigated the immunological phenotype associated with a novel hemizygous FLNA variant identified in a pediatric patient presenting with recurrent infections and inflammatory manifestations. Whole-exome sequencing revealed a hemizygous c.7405C>T (p.Pro2469Ser) variant in FLNA, which was confirmed by Sanger sequencing. Its potential impact on immune-cell function and cytoskeletal organization was evaluated using confocal microscopy, flow cytometry, and molecular assays. Patient-derived T cells showed impaired activation and proliferation following CD3/CD28 and IL-2 stimulation, accompanied by reduced CD25 and CD69 upregulation. CD4+ T cells also exhibited reduced IFN-γ, TNF-α, and IL-2 production after stimulation. Despite elevated basal phospho-STAT5 levels, IL-2-induced STAT5 phosphorylation and TCR-associated signaling responses, including pZAP70, pLCK, and p38 MAPK activation, were attenuated. Confocal imaging together with image-based quantification demonstrated altered cortical cytoskeletal organization in patient T cells despite preserved FLNA expression. In parallel, NK cells showed impaired activation responses and reduced cytotoxic activity under the assay conditions used. Increased apoptosis was observed in CD4⁺, CD8⁺, and NK-cell populations. Inflammatory cytokines were elevated in plasma and colonic tissue, whereas colonic ZO-1 and FLNA expression were reduced. Collectively, these findings indicate that the FLNA p.Pro2469Ser variant is associated with altered immune-cell signaling, disturbed cortical cytoskeletal organization, and immune dysregulation. This study expands the phenotypic spectrum linked to FLNA variants and supports a role for filamin A in human immune-cell regulation.
HTLV-1 infection causes chronic immune activation and a prolonged asymptomatic phase, but the functional features that define early disease remain unclear. We evaluated whether spontaneous proliferation (SP) and related functional immune parameters can distinguish clinical stages across the HTLV-1 spectrum. Spontaneous and mitogen-induced lymphoproliferation (PHA, anti-CD3), measured by CFSE dilution, and in vitro cell death, measured by cytometry, were assessed in 435 HTLV-1-infected individuals: asymptomatic carriers (AC, n = 303), individuals with intermediate syndrome (IS, n = 21), patients with HTLV-1-associated myelopathy (HAM, n = 94), adult T-cell leukemia/lymphoma (ATL, n = 17), and two control groups. Notably, SP levels divided the asymptomatic individuals in two subgroups: low-SP (24.5
This study aimed to identifydistinct profiles of response to Tumor Necrosis Factor inhibitors (TNFi) in rheumatoid arthritis (RA) patients using unsupervised machine learning to integrate clinical, demographic, and genetic data. A cohort of 294 RA patients was analyzed using hierarchical clustering techniques. Data collected included body mass index (BMI), adherence to physical activity, prevalence of comorbidities, seropositivity, Health Assessment Questionnaire (HAQ) scores, and genetic polymorphisms in TNF pathway. Responders to TNFi demonstrated a more favorable clinical profile, with a lower BMI (25.6 vs. 27.0; p = 0.03), higher physical activity adherence (56.6
Recurrent pregnancy loss (RPL) is a multifactorial reproductive disease that is typified by repeated pregnancy loss with immune dysregulation emerging as a critical contributing factor. Immunoglobulins are among the immune components that are instrumental in preserving tolerance between the mother and the fetus, regulating inflammatory responses, and supporting placental development. This review provides a comprehensive overview of the role of immunoglobulins in recurrent pregnancy loss, with particular emphasis on immune dysregulation, emerging biomarkers, and therapeutic implications. The immunoglobulin profiles, such as the changes in IgG subclasses and the presence of pathogenic autoantibodies, e.g. antiphospholipid and antinuclear antibodies, are highly correlated with unfavourable pregnancy outcomes. These changes, in a mechanistic manner, lead to poor trophoblast invasion, endothelial dysfunction, thrombosis and cytokine imbalance. Emerging evidence also highlights the contribution of regulatory B cells and the antibody-mediated immune network in maintaining pregnancy homeostasis. Immunoglobulin profiling has clinical potential as a diagnostic and prognostic methodology that can be used to stratify patients and provide customised therapy. Immunomodulatory therapies, especially intravenous immunoglobulin (IVIG), are potentially effective in a limited range of patient populations, but clinical response is variable. On the whole, this review demonstrates the role of immunoglobulin-mediated mechanisms in RPL and their future potential in enhancing precision medicine approaches to better diagnosis and management.
Chronic viral coinfections may contribute to persistent immune dysregulation in people with HIV, but their association with PD-1/PD-L1 expression remains uncertain. We analysed 100 people with HIV using available-case endpoint analyses and marker-specific paired baseline-to-follow-up analyses. Follow-up was scheduled at 12 months (allowable window, 9–15 months), and all participants had HIV RNA < 50 copies/mL at the endpoint. Baseline HBV contact, HCV status, and CMV IgG serostatus were evaluated in relation to PD-1 and PD-L1 expression on CD4⁺ T cells and CD19⁺ B cells and exploratory composite checkpoint-expression indices. HBV contact and CMV IgG serostatus were not associated with reproducible differences. The global comparison of longitudinal CD4⁺ T-cell PD-L1 change across HCV groups did not remain significant after false-discovery-rate correction; an exploratory pairwise comparison suggested a greater increase in participants with baseline HCV RNA positivity. Overall, the HCV-related pattern was hypothesis-generating and requires prospective validation.
This study aims to assess the circulating galectin-3 (Gal-3) levels in patients with systemic lupus erythematosus (SLE) and explore the major related influencing factors. Systematic searches of all relevant English articles were conducted in PubMed, Embase, Web of Science, and the Cochrane Library from inception to September 22nd, 2025. Continuous variable data from multiple studies were subjected to a random-effects meta-analysis using Stata 12.0 software to estimate the pooled effect size. Data on circulating Gal-3 levels were analyzed using the standard mean difference (SMD) with 95
Pustular psoriasis (PP), including generalized pustular psoriasis (GPP) and palmoplantar pustulosis (PPP), is a rare and severe inflammatory dermatosis distinct from plaque psoriasis and associated with significant unmet therapeutic needs. Although advances in immunopathogenesis have identified key cytokine and signaling pathways, evidence-based treatment options remain limited. This systematic review evaluates the efficacy and safety of emerging Janus kinase (JAK) and tyrosine kinase (TYK) inhibitors in the management of PP. This systematic review was conducted in accordance with PRISMA 2020 guidelines. PubMed/Medline, Ovid-Embase, and Web of Science were searched from inception to November 15th, 2025, to identify all English-language clinical studies evaluating JAK or TYK inhibitors in patients with GPP or PPP. Methodological quality and risk of bias were independently assessed using National Institutes of Health quality assessment tools and the Murad et al. criteria. Of 2,259 records identified, 33 clinical studies (177 patients with GPP or PPP) met the predefined eligibility criteria. TYK inhibitors were evaluated in three studies of deucravacitinib (n = 11), which showed variable efficacy across reports: improvements in PASI/PPPASI, symptoms, and quality of life in some patients, and discontinuation due to insufficient response in others, with no serious adverse events reported. JAK inhibitors were evaluated in 30 studies involving 166 patients and were associated with improvements in disease severity, quality of life, and physician-assessed outcomes. Favorable responses were reported in observational studies, while rapid clinical improvement was frequently described in case-based evidence. Overall, treatment was generally well tolerated, although interpretation is limited by the predominance of uncontrolled studies and heterogeneous data. Available evidence indicates that JAK and TYK inhibitors may provide clinical benefit in GPP and PPP, particularly in refractory cases, with generally acceptable safety profiles. However, conclusions are limited by small, heterogeneous studies, and well-designed randomized controlled trials with longer follow-up are needed to establish long-term efficacy and safety.
Anti-topoisomerase I (anti-Scl-70) antibodies are strongly associated with systemic sclerosis and fibrotic organ involvement. However, their significance in ANA-positive individuals without established systemic sclerosis remains unclear. We investigated whether isolated anti-Scl-70 positivity is associated with subclinical cardiac involvement. This retrospective single-center study included 486 ANA-positive individuals identified between 2016 and 2026. Patients were classified as anti-Scl-70-positive (n = 242) or anti-Scl-70-negative (n = 244). Individuals with systemic sclerosis, other rheumatologic diseases, cardiovascular disease (except hypertension), malignancy, or advanced renal/hepatic disease were excluded. All anti-Scl-70-positive patients remained free of systemic sclerosis during at least 12 months of follow-up. Electrocardiographic Diastolic Index-1 (EDI-1) and EDI-2 were calculated from standard 12-lead ECGs. Demographic characteristics, inflammatory markers, ANA titers, and most ENA antibodies were similar between groups. Anti-Ro52 positivity was more frequent in anti-Scl-70-positive individuals (p = 0.004). EDI-1 was significantly lower, whereas EDI-2 was significantly higher in the anti-Scl-70-positive group (both p < 0.001). Among patients with available echocardiographic data, anti-Scl-70-positive individuals demonstrated higher E/e′ ratios (p = 0.004) and lower LV-GLS values (p = 0.046). EDI-1 showed a strong correlation with E/e′ (rs = -0.898, p < 0.001). In multivariable analysis, anti-Scl-70 positivity was the only independent predictor of electrocardiographic diastolic dysfunction (p < 0.001). Isolated anti-Scl-70 positivity is associated with impaired electrocardiographic markers of diastolic function in ANA-positive individuals without overt systemic sclerosis. These findings suggest that isolated anti-Scl-70 positivity is associated with early subclinical myocardial involvement and support further investigation of ECG-derived diastolic indices as potential screening tools in this population.
BACKGROUND:Retinal ischemia-reperfusion (RI/R) injury causes vision loss and lacks effective treatments. The role of diosmetin (DIO) in this condition, particularly through the SIRT1/Nrf2 pathway and its effect on PANoptosis, is unknown. METHODS:An in vitro oxygen-glucose deprivation/reperfusion (OGD/R) model was developed using R28 retinal precursor cells, while an in vivo rat model of retinal ischemia/reperfusion (RI/R) was created through transient intraocular pressure elevation. DIO was administered in vitro, at the onset of reperfusion, and in vivo, via intraperitoneal injection. Cell viability, proliferation, oxidative stress markers, inflammatory cytokines, and PANoptosis-related protein expression were assessed. The role of SIRT1 was confirmed using siRNA knockdown in vitro and the pharmacological inhibitor EX527 in vivo. RESULTS:DIO treatment notably enhanced cell viability and proliferation in OGD/R-injured R28 cells, while also maintaining retinal structure and neuronal survival in RI/R-injured rats. DIO stimulated the SIRT1/Nrf2 pathway, evidenced by elevated levels of SIRT1, nuclear Nrf2, and HO-1 expression. This activation diminished oxidative stress, evidenced by lower ROS and MDA levels and higher SOD, CAT, and GSH levels, while also reducing inflammation, as indicated by decreased TNF-α, IL-1β, and IL-18. Consequently, DIO inhibited PANoptosis by downregulating key markers of apoptosis (cleaved caspase-3, BAX), pyroptosis (NLRP3, GSDMD), and necroptosis (p-RIPK3, p-MLKL). The beneficial effects of DIO, including neuroprotection, antioxidant activity, anti-inflammation, and anti-PANoptosis, were entirely nullified by SIRT1 knockdown or inhibition. CONCLUSION:DIO protects against retinal I/R injury by activating SIRT1/Nrf2 to inhibit oxidative stress, inflammation, and PANoptosis, highlighting its therapeutic potential.
Chronic urticaria is a common mast-cell-driven inflammatory disorder characterized by recurrent wheals, angioedema, or both. Angioedema is associated with higher disease burden, impaired quality of life, and greater difficulty in clinical assessment, yet the overall research structure of chronic urticaria in the context of angioedema has not been systematically mapped. Publications were retrieved from the Web of Science Core Collection on July 14, 2026. English-language Articles and Reviews published between 1947 and 2026 were included after independent screening by two researchers, with disagreements resolved by a third researcher. Bibliometrix, VOSviewer, CiteSpace, Scimago Graphica, and GraphPad Prism were used to analyze annual output, countries, institutions, authors, journals, co-cited references, and keyword evolution. A total of 881 publications were included. Annual output increased over time, with faster growth after 2014. The United States ranked first in productivity, Germany had the highest citation count and total link strength, and Charité – Universitätsmedizin Berlin was the leading institution. The knowledge base was concentrated in mast cell biology, autoimmune mechanisms, anti-IgE therapy, and patient-reported outcomes. Keyword analysis showed a shift from early immune plausibility and idiopathic descriptions toward disease standardization, quality of life, and targeted therapy, including omalizumab and newer targeted treatments. Angioedema remained a high-frequency term and was repeatedly linked to disease burden, assessment, and treatment. The literature addressing both chronic urticaria and angioedema showed increasing emphasis on structured clinical assessment, mast-cell and autoimmune mechanisms, patient-reported outcomes, biomarker-oriented stratification, and targeted treatment. Future research should broaden international participation and evaluate clinically useful biomarkers and patient-centered outcomes across diverse populations.
Background: Antinuclear antibodies (ANAs) are serological markers of clinical importance in the diagnosis and monitoring of autoimmune diseases. This study aimed to develop a HEp-2 cell-based ELISA method as a quantitative and standardized alternative to traditional ANA detection techniques such as indirect immunofluorescence (IFA). Materials and methods: HEp-2 cells were seeded in ELISA plates at 3.0 × 10⁴ cells per well and incubated. After fixation with formaldehyde and acetone, cells were quenched with 1
