The NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome is an established driver of inflammation in diseases such as diabetes, Alzheimer's disease, and gout. Previously, we screened 875 FDA-approved drugs for NLRP3 inhibitors and identified ponatinib as one of the five candidates that reduced NLRP3 inflammasome activation without causing cytotoxic effects in bone marrow-derived macrophages (BMDMs). Therefore, we hypothesize that ponatinib may be an effective NLRP3 inflammasome inhibitor. We performed dose curves and cytotoxicity assays to determine an effective in vitro concentration of ponatinib in BMDMs (1 μM) and primary microglia (0.5 μM) that reduced IL-1β secretion without inducing cytotoxicity. In BMDMs, ponatinib inhibited Nlrp3- and Caspase-1-dependent IL-1β and IL-18 secretion and significantly reduced caspase-1 processing. Ponatinib also reduced IL-1β secretion by microglia, indicating possible NLRP3 inflammasome inhibition in this cell type, but further testing is required. Although ponatinib has previously been demonstrated to drive cardiotoxicity-mediated inflammation, these data suggest that at specific doses, in vitro ponatinib can be non-cytotoxic and effective in attenuating NLRP3 inflammasome activation in macrophages and microglia.
N6-methyladenosine (m6A) modification plays a crucial role in regulating immune responses against Mycobacterium tuberculosis (Mtb) infection, but the underlying mechanisms remain unclear. Here, we analyzed m6A regulatory gene expression in GEO dataset GSE139825 and validated the finding in THP-1-derived macrophages. Bioinformatics analysis showed that METTL14, ALKBH5, and YTHDC1 were significantly downregulated, while METTL3 was upregulated in Mtb-infected alveolar macrophages compared to controls. In vitro experiments confirmed METTL14 expression was dose-dependently reduced with Mtb infection in macrophages, and METTL14 overexpression reversed Mtb-induced decreases in cell viability, reduced LDH release, and diminished intracellular Mtb burden. Mechanistically, METTL14 overexpression restored Mtb-suppressed autophagy, evidenced by increased LC3II/I ratio and Beclin1 expression as well as decreased p62 levels, which was abrogated by the autophagy inhibitor 3-MA. Further, as validated by bioinformatic prediction, RIP assay and actinomycin D experiment, METTL14 directly bound to TAX1BP1 mRNA and enhanced its m6A modification, thereby stabilizing TAX1BP1 mRNA and upregulating its expression. TAX1BP1 knockdown abolished the protective effects of METTL14 overexpression on macrophage viability, autophagy, and anti-Mtb capacity. Collectively, our findings demonstrate that METTL14, downregulated by Mtb infection, promotes macrophage autophagy and antibacterial activity via m6A-mediated stabilization of TAX1BP1. This METTL14-TAX1BP1-m6A pathway provides a novel therapeutic target for tuberculosis treatment.
PURPOSE:This study evaluated the association of combining veno-venous extracorporeal membrane oxygenation (ECMO) with whole-lung lavage (WLL) and granulocyte-macrophage colony-stimulating factor (GM-CSF) nebulization in pulmonary alveolar proteinosis (PAP), focusing on pulmonary function, oxygenation, lipid metabolism, and inflammatory modulation. METHODS:This retrospective cohort study included 40 PAP patients, categorized into WLL + GM-CSF (n = 20) and ECMO + WLL + GM-CSF (n = 20) groups according to the treatment they received. Outcomes included pulmonary function [diffusing capacity for carbon monoxide (DLCO%), forced vital capacity (FVC%)], arterial blood gas parameters [partial pressure of oxygen (PaO2), carbon dioxide (PaCO2), oxygen saturation (SaO2)], lipid profiles [total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C)], inflammatory cytokines such as interleukin-6 (IL-6) and C-reactive protein, and functional assessments (Barthel Index score, 6-min walk distance). Correlations between lipids profiles, arterial blood gas parameters, pulmonary functional indicators, and inflammatory cytokines were evaluated using Pearson's or Spearman's correlation coefficients. RESULTS:The ECMO + WLL + GM-CSF group showed greater improvements in DLCO% and FVC%, significant increases in PaO2 and SaO2, and greater reductions in PaCO2, pro-inflammatory markers (IL-6, CRP), and lipid levels (TC, TG, LDL-C) compared to the WLL + GM-CSF group. In the ECMO + WLL + GM-CSF group, negative correlations were observed between serum TG and PaO2, DLCO%, and FVC%, and IL-6 inversely correlated with FVC%. The ECMO + WLL + GM-CSF group also achieved higher functional recovery based on the higher Barthel index scores and longer 6-min walk distance. CONCLUSION:ECMO-assisted WLL combined with GM-CSF was associated with greater improvements in pulmonary function and oxygenation, as well as greater reductions in serum lipid levels and inflammatory markers.
BACKGROUND:Interferon (IFN) is an important treatment for chronic hepatitis B (CHB), with highly variable clinical efficacy, and how IFN-α mediates its therapeutic benefits via innate immunity, particularly macrophages, is not fully understood. OBJECTIVES:To elucidate whether and how the CCL2-CCR2-macrophage signaling axis mediates the antiviral efficacy of interferon-α therapy in chronic hepatitis B. DESIGN:We first analyzed the hepatic immune microenvironment induced by long-term IFN-α treatment using a public single-cell dataset (GSE237519). Next, we utilized an AAV-HBV-infected type I interferon receptor humanized mouse model (huIFNAR mice) and treated the mice with short-term pegylated interferon-α-2b (PegIFNα-2b), combined with either CCR2 antagonism (INCB3344) administration or macrophage depletion via clodronate liposomes. Through single-cell RNA sequencing (scRNA-seq), flow cytometry, immunohistochemistry, and serum HBV biomarker detection, we systematically investigate the role of the CCL2-CCR2-macrophage signaling axis in the anti-HBV effect of interferon. RESULTS:Long-term IFN treatment reshaped the hepatic macrophage landscape and activated CCL signaling. Depletion of macrophages attenuated IFN-mediated suppression of HBV DNA replication and impaired the decline in HBsAg levels. CCR2 antagonism impaired the antiviral efficacy of PegIFNα-2b and inhibited monocyte recruitment to the liver. Single-cell RNA sequencing further revealed that the CCR2 antagonist disrupted the CCL2-CCR2 signaling axis, which in turn arrested Ly6c + macrophage differentiation, disrupted intercellular communication networks, and promoted a more pronounced pro-inflammatory state. CONCLUSIONS:Our study defines the CCL2-CCR2-Ly6c+ macrophage axis as a critical immunoregulatory pathway in IFN therapy for CHB. This axis coordinates macrophage recruitment, functional differentiation, and cellular crosstalk to mediate the antiviral effects of PegIFNα-2b. These findings may highlight a potential target to refine IFN-driven immunotherapies for CHB treatment.
Objective This study investigated the impact of cigarette smoke extract (CSE) on BEAS-2B bronchial epithelial cells, focusing on whether it induces pyroptosis via the ROS/NLRP3/GSDMD pathway and how NAC protects these cells from pyroptosis. Methods BEAS-2B cells were treated with varying CSE concentrations, and cellular damage was evaluated via PI staining and release of ROS and LDH. Cellular morphology alterations were examined by immunohistochemistry, and IL-18 and IL-1β release were measured by ELISA. Western blot analysis was used to assess expression of proteins linked to pyroptosis, including ASC, NLRP3, cleaved-caspase-1, GSDMD-N, and IL-1β. Caspase-1 and GSDMD inhibitors were utilized to elucidate the underlying mechanisms. Results Following CSE treatment, bronchial epithelial cells demonstrated inflammatory infiltration and membrane rupture and increased ROS, NLRP3, cleaved-caspase-1, GSDMD-N, IL-18, and IL-1β expression. Treatment with caspase-1 and GSDMD inhibitors decreased expression of these markers, as expected. Interestingly, NAC treatment reduced the expression of these proteins compared to the CSE group, suggesting a protective role against pyroptosis through inhibition of the ROS/NLRP3/GSDMD pathway. Conclusion The study shows that CSE triggers pyroptosis in COPD via the ROS/NLRP3/GSDMD pathway, with NAC offering antioxidative protection. These findings enhance understanding of AECOPD pathophysiology and support NAC's therapeutic role in treatment.
Background and purpose: Intrauterine adhesion (IUA) is a prevalent gynecological disorder that causes infertility and recurrent miscarriage; however, effective treatments remain limited. This study aimed to elucidate the function of Sirt3 in IUA and explore its underlying molecular mechanisms. Methods: A rat IUA model was established by simulating mechanical endometrial injury, and an in vitro model was induced by treating human endometrial epithelial cells (hEECs) with recombinant TGF-β1. The expression of miRNAs and key proteins was detected via RT-qPCR, Western blotting, immunohistochemistry, and immunofluorescence. Cytokine levels were measured via ELISA, and endometrial damage in rats was assessed by using hematoxylin and eosin (HE) and Masson staining. Results: We observed significant Sirt3 downregulation in IUA. Sirt3 overexpression ameliorated endometrial damage and fibrosis in IUA rats, thereby suppressing the expression of epithelial-mesenchymal transition (EMT) markers (vimentin and N-cadherin) and fibrosis-related proteins (α-SMA and collagen I) while also restoring E-cadherin expression. The overexpression of Sirt3 could also promote autophagy in hEECs induced by TGF-β1 (which promotes the expression of LC3 and Beclin 1 and inhibits the expression of p62), inhibit pyroptosis (which suppresses the expression of caspase-1, NLRP3, GSDMD, and ASC), and downregulate the levels of IL-1β and IL-18. These effects were reversed by the autophagy inhibitor chloroquine or the pyroptosis activator nigericin. Further investigation revealed that miR-1-3p was upregulated in IUA and could directly target and negatively regulate Sirt3 expression. Treatment with miR-1-3p antagomir suppressed EMT and fibrosis in hEECs under TGF-β1 exposure; however, this effect was attenuated by concomitant Sirt3 knockdown. Conclusion: In summary, miR-1-3p promotes IUA progression by suppressing Sirt3-mediated autophagy and activating pyroptosis, thereby driving EMT and fibrosis. The findings of this research revealed that the key role of Sirt3 in inhibiting the progression of IUA provides a potential target for the development of disease intervention therapies.
The effects of long-term parental microplastic exposure on offspring immunity remain unclear. This study investigated how different parental exposure patterns affect immune status in F1 offspring. Parental rats were divided into four groups: paternal, maternal, dual-parental, and control. Treated groups received polystyrene microplastics (5 mg/L) in drinking water for 90 days. F1 offspring were raised under standard conditions until 8 weeks of age, after which fecal samples, thymus, spleen, and serum were collected for immune evaluation. Maternal microplastic exposure significantly disrupted gut microbiota α-diversity, dysbiosis index, and composition in F1 offspring (all P < 0.05). These changes were accompanied by decreased RBCs and PLT counts, elevated serum TNF-α, reduced thymic CD3+ and CD4+ T cells, and downregulated IL-10 mRNA and NF-kB protein expression. Histological examination revealed blurred corticomedullary boundaries, sparse cellularity, and lymphocyte vacuolization in the thymus, along with thinning of the periarteriolar lymphatic sheaths in the spleen, further indicating immune imbalance. Paternal and dual-parental exposure also induced gut microbiota dysbiosis and reduced thymic CD4+ T cells. Paternal exposure upregulated Th17-related RORγt and TNF-α mRNA in the thymus, whereas dual-parental exposure decreased thymic Nrf2 protein, splenic CD4+/CD8+ ratios, and white pulp area. Thymic pathology was observed in both groups. In conclusion, our findings suggest that prolonged parental exposure to microplastics may disrupt immune homeostasis in F1 offspring by altering gut microbiota composition and modulating oxidative stress or inflammatory responses. Moreover, the extent of these effects varies with the different parental exposure patterns.
Coronaviruses (CoVs) cause severe respiratory diseases and continue to pose a significant worldwide health threat. Antibodies that specifically target the structural proteins of SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) are crucial for immunological research, development of diagnostic assays, and evaluation of immune responses. Avian egg yolk immunoglobulin Y (IgY) represents a novel approach that has attracted considerable interest due to its strong immunological response, scalability, and excellent safety profile. The study involved generating SARS-CoV-2 receptor-binding domain (RBD)-specific spike (S) IgY antibodies by immunising laying hens with a synthetic RBD epitope. IgY antibodies were extracted from egg yolks using optimised purification methods and subsequently characterised for yield, purity, and antigen specificity. The purified IgY preparations were evaluated and verified for specificity and binding affinity to the target antigen using indirect enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), conventional Western blotting, and automated Jess Western blot analysis. The produced antibodies exhibited targeted recognition of the antigen and were effectively purified with commendable yield and purity, highlighting their potential as a cost-effective, scalable, and dependable source of primary antibodies for immunological research and diagnostic applications. Additional research, including virus-neutralisation and in vivo efficacy assessments, is needed to explore their potential for passive immunisation and various biomedical applications.
Background Diabetic foot ulcers (DFU) are one of the most serious complications of diabetes, closely associated with high amputation rates, mortality, and healthcare burdens, and the clinical treatment research is still limited. Negative Pressure Wound Therapy (NPWT) is a treatment modality for DFU, but its specific mechanisms promoting wound healing remain largely unknown. Methods The bulk RNA-seq and scRNA-seq data for DFU used in this study were obtained from the GEO database. First, this study used differential analysis and multiple machine learning algorithms to identify key genes activated by NPWT. Subsequently, this study used GSVA and AddModuleScore analysis to evaluate the mesenchymal-epithelial transition (MET) capability of keratinocytes (KCs), and used CytoTRACE and pseudotime trajectory analysis to construct MET differentiation trajectories. Finally, immunofluorescence (IF) was used to validate the expression of GRHL2 in DFU clinical samples. Results Differential expression analysis identified 975 dysregulated genes in DFU (530 upregulated, 445 downregulated). Through four machine learning algorithms, GRHL2 was consistently selected. GRHL2 was primarily expressed in skin at both RNA and protein levels. Single-cell analysis of 13 DFU samples confirmed GRHL2 is predominantly expressed in KCs. Specifically, NPWT activates GRHL2, thereby driving mesenchymal fibroblasts (MFs) into supraspinous_KCs. Additionally, molecular docking suggests that Parthenolide, MG-132, Mitoxantrone, and Irinotecan may have potential efficacy in treating DFU. Finally, IF results show that GRHL2 expression is consistent with transcriptomic analysis results. Conclusion Overall, this study established a MET model of “MFs - supraspinous_KCs” in DFU repair. Additionally, the “NPWT - GRHL2 - MET - cell conversion” signaling axis proposed in this study supplements the molecular mechanism by which NPWT promotes wound healing through the regulation of cell fate determination, deepening the biological effects of physical stimulation from the macroscopic tissue repair phenotype to the microscopic cell conversion level, and further enriching the mechanism by which NPWT promotes wound healing.
Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder in which immune dysregulation has emerged as an important component of disease pathogenesis; however, the contribution of circulating proteins and their cellular context remains incompletely understood. Here, we performed an integrative multi-omics analysis combining Mendelian randomization (MR), bulk transcriptomics, single-cell RNA sequencing, and peripheral blood validation to systematically identify plasma proteins associated with AD.Proteome-wide MR analysis identified multiple circulating proteins associated with AD risk. Integration with transcriptomic data identified AIF1 (allograft inflammatory factor 1) as a shared candidate supported by both genetic prioritization and differential expression analysis. Although bulk transcriptomic data showed reduced AIF1 expression in AD, single-cell analysis revealed distinct cell type-specific expression patterns, with predominant enrichment in monocytes and other innate immune populations. PBMC-based qPCR further confirmed an overall reduction in AIF1 expression in AD.Further analyses suggested that AIF1-associated immune alterations were linked to changes in inflammatory signaling pathways, including STAT, IRF, and NF-κB-related activity, as well as differences in intercellular communication involving MIF, GALECTIN, ANNEXIN, and CypA-related signaling. Peripheral immune cell composition analysis indicated differences between AD and control samples, characterized by relative changes in innate immune cell proportions.Collectively, these findings identify AIF1 as an immune-associated factor linked to genetic and transcriptional alterations in AD and suggest its association with monocyte-related immune states and altered immune signaling patterns. This study provides a multi-layered framework for investigating peripheral immune involvement in AD and highlights potential directions for understanding immune-related alterations and biomarker discovery.
T cell fate is partly determined by T cell receptor (TCR) affinity for its cognate peptide ligand. During thymic selection, CD4+ T cells with higher affinity for self-antigens are more likely to be deleted or become FOXP3+ natural T regulatory (nTreg) cells. However, downstream transmembrane adaptor proteins such as the T cell receptor associated transmembrane adaptor 1 (TRAT1) also modulate T cell activation. Here we show that TRAT1 decreases the activation threshold of human naïve CD4+ T cells and mediates TCR activation leading to transient FOXP3 expression via FOXO3A. Individuals at high risk for autoimmune type 1 diabetes (T1D) had increased proportions of CD4+ T cells with elevated TRAT1 expression in their naïve CD4+ T cell compartment. Naïve CD4+ T cells transduced with an insulin-specific TCR-TRAT1 construct exhibited enhanced proliferation, even in the absence of exogenously added insulin peptide, and showed heightened sensitivity to low doses of insulin peptide. These findings reveal an additional layer of T cell reactivity regulation independent of TCR affinity, which may underlie homeostatic and pathogenic functions of human T cells.
Anti-neutrophil cytoplasmic antibody (ANCA) is an important serological marker for autoimmune diseases, particularly ANCA-associated vasculitis (AAV). AAV frequently presents with refractory rhinosinusitis as an initial manifestation, yet ANCA testing in patients with rhinosinusitis continues to face challenges, including inconsistent subtype classification, variable detection performance, and difficulties in early identification, leading to misdiagnosis and delayed treatment. This narrative review focuses on the diagnostic value of ANCA in AAV-related rhinosinusitis, encompassing granulomatosis with polyangiitis (GPA), microscopic polyangiitis (MPA), and eosinophilic granulomatosis with polyangiitis (EGPA). We summarize the research progress on ANCA detection and its association with clinical, imaging, and pathological features, aiming to provide a theoretical reference for early clinical diagnosis, individualized intervention, and improved patient prognosis.
BACKGROUND:Pancreatic cancer is among the most lethal cancers because of late diagnosis and inadequate therapeutic preferences. The tumor microenvironment and immune response have critical roles in disease progression and patient outcomes. LncRNAs are regulators of immune function and tumorigenesis, yet their prognostic value in pancreatic cancer remains partly clarified. METHODS:RNA-sequencing data and corresponding clinical features for 178 pancreatic cancer patients were obtained from TCGA. Immune-related mRNAs were retrieved from the MSigDB, and immune-related lncRNAs were identified based on a significant Pearson correlation with at least two immune-related mRNAs. Regression analyses were sequentially applied to construct a prognostic immune-related lncRNA set. The predictive performance of the set was assessed using time-dependent ROC curves and Kaplan-Meier survival analysis in both the TCGA training cohort and an independent external validation cohort (GSE224564). RESULTS:From 4059 lncRNAs, 1172 immune-related lncRNAs demonstrated strong associations with immune-related mRNAs. Sequential screening identified 34 survival-associated lncRNAs by univariate Cox regression (P < 0.01), 12 lncRNAs following LASSO regression, and ultimately two lncRNAs-CASC8 (HR = 1.7, P < 0.05) and LINC01004 (HR = 0.54, P < 0.05)-by multivariate Cox regression. A risk score signature was developed using these two lncRNAs. Time-dependent ROC analysis demonstrated moderate-to-good predictive precision in the training cohort, with area under the curve (AUC) values of 0.702, 0.774, and 0.84 for 1-, 3-, and 5-year overall survival, respectively. Patients in the high-risk group exhibited significantly worse survival compared to the low-risk group (P < 0.05). External validation in the GSE224564 cohort confirmed consistent risk stratification (P = 0.019), though with lower discriminative accuracy (AUC ∼ 0.60). Multivariate analysis demonstrated that the risk score acted as an independent prognostic factor in both cohorts. CONCLUSION:The CASC8 and LINC01004 immune-related lncRNA profile serves as a potential prognostic marker for pancreatic cancer. This set demonstrates increasing predictive trends over time in the training dataset. The presented findings offer novel insights into the correlation of immune-related lncRNAs with pancreatic cancer progression and provide a foundation for future risk stratification modeling.
BACKGROUND:Colorectal cancer (CRC) can induce stresses on the immune system that can affect both the numbers and function of immune cells. Changes in immune cell functions can also occur during ageing and these may affect both the ability to fight infections and to protect against cancers. As the incidence of CRC is age-related, the aim of this work was to identify changes in immune cell subtypes that are specific to CRC and not merely due to age-related changes. METHODS:Whole blood samples from 49 CRC patients about to undergo surgery and 22 healthy controls were collected. Samples were analysed by immunophenotyping, detection of HLA-DR on T-lymphocytes and monocytes, and senescent-like T-lymphocytes. RESULTS:CD4 staining intensity of monocytes was significantly increased in CRC patients and showed a positive correlation with their HLA-DR staining intensity. In most CRC patients, the numbers of helper T-lymphocytes were lower with the progression of the disease, while cytotoxic T-lymphocytes were higher (an opposite pattern to the immunophenotypes of the healthy ageing cohort). NKbright cells were higher while NKdim cells were lower in patients with large tumours. An increase in the T-lymphocytes to B-lymphocytes ratio correlated with the metastatic status. CONCLUSIONS:Complex changes in the immune phenotypes in CRC, distinct from those that occur during ageing were observed, that imply development of an immuno-suppressive phenotype that may aid tumour evasion of immunity.
OBJECTIVE:Concentrated growth factor (CGF) is rich in growth factors and complement proteins like C3a and C5a, showing promise for enhancing tissue regeneration. This study explores the regulatory effects of recombinant C3a/C5a proteins on the osteogenic differentiation of PDLSCs in an inflammatory microenvironment and their potential association with the Wnt/β-catenin pathway. METHODS:P-PDLSCs were generated by treating PDLSCs with IL-1β (5 ng/mL) and TNF-α (10 ng/mL) for 24 h. Subsequently, P-PDLSCs were treated with C5a and C3a, followed by evaluation of cell viability and early osteogenic differentiation capacity (ALP activity). To investigate the underlying mechanism, cells were treated with the Wnt/β-catenin pathway inhibitor, XAV939, either alone or combined with C3a/C5a. Cell viability, ALP activity, and the mRNA and protein levels of AKT, Runx2, and β-catenin were measured. RESULTS:Compared with the control group, exogenous recombinant C3a and C5a treatment can promote the proliferation and early osteogenic differentiation of P-PDLSCs. Additionally, through KEGG enrichment, the Wnt/β-catenin pathway was identified as a key target. The pro-osteogenic effects of C3a/C5a were partially reversed by XAV939, pointing to a potential correlation with the Wnt/β-catenin signaling pathway. CONCLUSION:Exogenous recombinant C3a/C5a is associated with increased early osteogenic differentiation of P-PDLSCs, a process correlated with the activation of the Wnt/β-catenin signaling pathway, providing potential insights into regenerative strategies for periodontal tissues.
Abdominal aortic aneurysm (AAA) is characterized by an inflammatory response that plays a crucial role in its pathogenesis. This study aimed to investigate the association between novel circulating inflammatory biomarkers and AAA. A total of 579 patients (240 with AAA and 339 with valvular heart disease as non-AAA controls) were enrolled from the Department of Cardiovascular Surgery at Zhongnan Hospital of Wuhan University between January 2020 and October 2024. Clinical characteristics and levels of circulating inflammatory markers were compared between the two groups. Multivariate logistic regression analyses indicated that male gender, lower HDLC, and higher neutrophil-to-HDL-C ratio (NHR), systemic inflammation response index (SIRI), and aggregate index of systemic inflammation (AISI) were independent but modest risk factors for AAA (P < 0.05). Furthermore, these biomarkers correlated with AAA severity. In conclusion, gender, HDLC, NHR, SIRI, and AISI serve as useful predictive factors for AAA. The nomogram prediction model demonstrated good discriminatory ability and may facilitate early diagnosis of AAA.
Acute myocardial infarction(AMI)is an important type of cardiovascular disease, which seriously threatens the lives of humans. In order to help patients receive timely clinical treatment and improve their survival rates, it is necessary to screen out related molecules in advance. To this end, we applied bioinformatics methods and machine learning algorithms to find possible biomarkers associated with AMI. When considering possible connections with dynamics of immune cells activation. Based on the RNA-seq results, differential expression analysis was performed followed by WGCNA to identify stably changed genes. The KEGG and GO enrichment analyses revealed that the identified genes are mainly related with inflammatory response, immune processes, apoptosis and immunomodulation. In our PPI network, some hub genes (e.g., FOS, JUN, TNF, IL1B, TLR2) were located in the center of the whole network. We found strong associations between the major genes and their interaction effects, we developed a predictive model for diagnosis using an artificial intelligence algorithm as follows: suggesting a substantially increased discriminative performance for AMI with the 4 hub genes NFKBIA, FCER1G, CD36 and ICAM1. The model achieved a high AUC score of 0.924. External validation based on an independent single-cell sequencing dataset demonstrated that NFKBIA, FCER1G, and ICAM1 displayed consistent expression patterns in both the training and test cohorts, with the highest expression levels observed in cardiomyocytes. Accordingly, these three genes may serve as reliable biomarkers for AMI and are closely associated with the distribution of immune cells.
BACKGROUND:Gastric cancer (GC) ranks as the fifth most prevalent malignancy globally. Emerging evidence implicates gut microbiome as a key modulator of anti-tumor immunity and immunotherapy response. Traditional Chinese Medicine (TCM) presents a promising yet underexplored avenue for microbiome modulation. METHODS:16S rDNA sequencing of fecal samples was used to detect changes of gut microbiota in advanced GC patients. GC models were established in huPBMC-NOG-dKO mice after fecal microbiota transplantation (FMT) to investigate the potential synergy between Number 3 Prescription (WD-3) and anti-PD-L1 monoclonal antibody (mAb) in treatment of non-responders. RESULTS:In this study, WD-3 combined with αPD-L1 showed additive benefit after the FMT of non-responders in the humanized mouse model model. WD-3 combination therapy correlated with reduced proportion of Treg cell infiltration in tumors. WD-3 combination was also associated with increased species richness and improved gut microbiota community structure compared to αPD-L1 alone, with increased relative abundances of Enterobacteriaceae and Lachnospiraceae. CONCLUSION:Our data provide correlative evidence that WD-3 supplementation combined with αPD-L1 treatment may attenuate GC progress in the FMT mouse model, indicating association with the modulation of gut microbiota. These findings aim to provide treatment strategies for the clinical treatment of advanced GC.
Gastrointestinal (GI) cancers continue to account for a major global cancer burden, with immune dysregulation recognized as a key determinant of their initiation and progression. Interleukin-27 (IL-27), a heterodimeric cytokine of the IL-12 family, has emerged as a pivotal but paradoxical regulator of tumor immunity in the GI cancers. IL-27 signals through the WSX-1/gp130 receptor complex, activating signal transducer and activator of transcription (STAT1) - and STAT3-dependent transcriptional pathways, yielding opposing outcomes. STAT1-dominant signaling promotes cytotoxic T-cell proliferation, T helper 1 (Th1) polarization, and natural killer (NK) cell cytotoxicity. However, STAT3-mediated responses induce IL-10, enhances regulatory T (Treg) cells suppression, stimulates myeloid-derived suppressor cells (MDSCs) activity, and upregulates programmed death ligand 1 (PD-L1), thereby facilitating immune evasion. These contrasting effects manifest in tumor-specific patterns: IL-27 demonstrates potent anti-tumor activity in experimental colorectal (CRC) and pancreatic cancer models, while elevated IL-27 associates with disease progression in hepatocellular carcinoma (HCC). Recent translational advances include IL-27-expressing viral vectors, dendritic cell platforms, and IL-27-blocking antibodies, underscoring its therapeutic duality. Overall, IL-27 functions as a context-dependent cytokine whose final effect on tumor progression is determined by the intrinsic characteristics of each tumor, the composition of tumor microenvironment (TME) and the interaction between immune and epithelial cells. Understanding the mechanisms that drive the dual roles of IL-27 across different GI cancers will be important for designing rational immunotherapeutic strategies that either enhance or inhibit IL-27 activity.
Defensins are key innate immune proteins exhibiting pleiotropic functions such as antibacterial activity, antiviral activity, anti-inflammatory activity, immunomodulatory functions, and roles in reproduction. Genomic information inadequately explains the whole gamut of the landscape of β-defensin in Bubalus bubalis and Ovis aries genomes. This study investigates the sequence evaluation and evolutionary relationships of β-defensin with 131 complete sequences in buffalo, sheep, and human. Sequence and domain-based analyses revealed diverse patterns of disulfide bridges and multiple signature patterns, including N- and O-glycosylation sites, N-myristoylation sites, and protein kinase C phosphorylation sites, indicating extensive post-translational regulation involved in various antimicrobial activities, signaling pathways, and reproduction. Phyre2-based 3D structural modeling, followed by PyMOL visualization, revealed β-strands stabilized by cysteine disulfide bonds and a short N-terminal α-helix. Variation analysis of β-defensin sequences identified unique allelic variants with several conserved amino acid positions. All sequences shared the characteristic six conserved cysteine residues, along with conserved glycine (G) in the GXC motif and glutamic acid (E), which contribute to structural stability and proper protein folding. We report sixteen amino acid sites depicting 9 distinct types of mutations within the cysteine residues, with most frequent substitutions of C→S (5/16) and C→P (3/16) amino acids. Phylogenetic and domain-based analyses across humans, sheep, and buffalo revealed eight major clusters, of which cluster I was found to be most diverse (36/131). Altogether, we provide a comprehensive analysis of β-defensin protein sequences revealing conserved features, functional motifs, and evolutionary relationships underpinning the functional role of β-defensins.
