Search Results (940)

Juvenile colorectal polyps (JP) are the most common polypoid lesions of the colon in children and the leading cause of lower gastrointestinal bleeding in pediatric patients. Although histologically classified as benign hamartomatous lesions, they arise in a context of allergic sensitization and IgE-mediated inflammation, suggesting that the epithelial compartment may play an active role in their pathogenesis. To date, most studies have focused on the immune cell infiltrate, leaving the molecular programs operating within the epithelium largely unexplored. Whole-transcriptome RNA sequencing (RNA-seq) was performed on epithelial cells isolated from pediatric JP (n = 8) and paired tissue circumjacent to polyp (TCP) (n = 7) obtained from children with a history of rectal bleeding and IgE sensitization to food allergens. Differential expression, functional enrichment (GO, KEGG, GSEA), and cross-dataset comparison with TCGA colorectal adenocarcinoma (CRC) was conducted. Candidate genes were validated by RT-qPCR in independent samples, including colorectal cancer (CRC) and inflammatory bowel disease (IBD) tissue. Differential expression analysis identified 3273 differentially expressed genes (2342 upregulated, 931 downregulated in JP vs. TCP), with 220 genes exceeding 32-fold change, including SERPINB3 (log2FC = 12.16), MMP1 (log2FC = 10.19), NMUR2 (log2FC = 10.08) and CHI3L1 (log2FC = 8.87). JP epithelium displayed a coherent type 2 inflammatory signature, encompassing upregulation of the alarmin IL33, eosinophil-attracting chemokines (CCL11, CCL24), IgE receptor subunits (FCER1A, FCER1G), and a coordinately activated leukotriene and prostaglandin biosynthetic program (ALOX5, ALOX5AP, PTGS2). Concurrent alterations in epithelial identity were observed, including downregulation of absorptive enterocyte and intestinal stem cell markers (CDX2, LGR5, ASCL2) alongside upregulation of secretory and regenerative programs, including the ectopic gastric-type mucin MUC5AC. Tight junction dysregulation—notably upregulation of the pore-forming claudin CLDN2 and loss of barrier-sealing claudins (CLDN3, CLDN4, CLDN23)—was consistent with impaired epithelial permeability. Pathway analyses confirmed activation of type 2 immune and extracellular matrix remodeling programs, with concurrent suppression of mitochondrial oxidative phosphorylation. Cross-dataset comparison with TCGA CRC data identified 381 co-upregulated and 87 co-downregulated genes shared between JP and CRC, including SERPINE1, CXCL8, ICAM1, and MMP3, several of which were associated with poorer disease-specific survival in CRC patients. RT-qPCR validation confirmed elevation of CHI3L1 and SERPINE1 in both JP and CRC, while SERPINB4 appeared JP-specific. Epithelial cells from pediatric juvenile colorectal polyps associated with allergic sensitization display a comprehensive type 2 inflammatory transcriptome alongside profound alterations in lineage identity, barrier integrity, and metabolic programming. Partial convergence with CRC-associated gene expression programs—in the absence of histological dysplasia—suggests that chronic allergic inflammation activates conserved epithelial remodeling pathways shared across mucosal tissues. CHI3L1, SERPINE1, and SERPINB4 are candidate biomarkers warranting validation in larger cohorts. Full article

The G protein-coupled receptor, CXC chemokine receptor 5 (CXCR5), is predominantly expressed on B cells located in the secondary lymphoid tissues, follicular helper T cells, and lymphoma cells. Binding to its ligand, CXCL13, mediates cell migration and regulates lymphocyte trafficking. Aberrant CXCL13/CXCR5 expression and signaling have been implicated in tumor progression, autoimmune diseases, and chronic inflammatory disorders. Therefore, specific mAbs against CXCR5 are expected to be useful for diagnosis and therapeutic applications. In this study, novel anti-human CXCR5 mAbs (Cx₅Mabs) were developed through flow cytometry-based high-throughput screening. One clone, Cx₅Mab-6 (IgG_2b, κ), recognized CXCR5-overexpressed Chinese hamster ovary (CHO)-K1 cells but did not react with the other five CXCR receptors-overexpressed CHO-K1 cells in flow cytometry. Additionally, Cx₅Mab-6 recognized endogenous CXCR5 in the human Burkitt lymphoma Raji cell line. The dissociation constant (K_D) values of Cx₅Mab-6 for CHO/CXCR5 and Raji were 3.4 × 10⁻⁹ M and 1.2 × 10⁻¹⁰ M, respectively. Furthermore, Cx₅Mab-6 is useful for Western blotting and can detect CXCR5 in human lymphoma tissue by immunohistochemistry. These findings suggest that Cx₅Mab-6 is versatile for basic research and has potential applications in clinical diagnosis. Full article

Skin aging is characterized by epidermal atrophy, reduced keratinocyte proliferation, and the accumulation of senescent cells that sustain a chronic, low-grade inflammatory secretome known as the senescence-associated secretory phenotype (SASP). We previously showed that Y-box binding protein 1 (YBX1) limits keratinocyte senescence in human epidermis by acting as a translational repressor of SASP chemokines, including CXCL1 and IL8. How this brake is regulated during skin aging, however, remains undefined. Here we demonstrate that, although total YBX1 protein is reduced in keratinocytes from aged human epidermis, the fraction of phosphorylated YBX1 (pYBX1) is increased relative to young donors, resulting in an elevated pYBX1/total YBX1 ratio that correlates with chronological age. Because pYBX1 is predominantly nuclear, whereas unphosphorylated YBX1 is cytoplasmic, this shift is predicted to deplete the cytoplasmic pool available for translational repression of CXCL1 and IL8. Consistent with this model, treatment of immortalized (Ker-CT) and primary human keratinocytes (HK) with the PI3K inhibitors PI-103 and GDC-0941 suppressed YBX1 phosphorylation without altering total YBX1 abundance, thus retaining YBX1 in the cytoplasm. This allows cytoplasmic YBX1 to re-engage and repress the translation of CXCL1 and IL8 mRNAs, lower chemokine secretion, and reduce the fraction of senescent keratinocytes. Moreover, conditioned medium from PI-103- or GDC-0941-treated cells sufficiently decreased senescence in recipient cells, consistent with a paracrine, secretome-mediated effect. Together, these findings support an association between YBX1 phosphorylation and SASP chemokine output in aging human keratinocytes and nominate YBX1-directed modulation as a candidate strategy for selectively attenuating senescence-associated secretory phenotypes in intrinsic skin aging. Full article

Background/Objectives: Osteosarcoma (OS) is an aggressive bone malignancy with a complex tumor microenvironment (TME) that influences therapeutic outcomes and resistance. How neoadjuvant chemotherapy (NACT) reshapes the OS TME at single-cell resolution remains largely undefined. This study aimed to characterize cellular heterogeneity in the OS TME after NACT and identify chemokine-mediated intercellular crosstalk driving chemoresistance. Methods: Single-cell RNA sequencing was performed on surgical specimens from 12 OS patients (6 treatment-naive and 6 post-NACT). After quality control, 77,616 cells (36,214 from naive patients, 41,402 from post-NACT samples) were analyzed through the Seurat pipeline. Unsupervised clustering, differential expression analysis, and cell–cell communication network construction were performed, and candidate signaling axes were validated using transwell assays and Western blotting. Results: Cells were classified into 10 major cell types. Osteoblasts, identified as malignant cells, were partitioned into 11 subpopulations with marked transcriptional heterogeneity and differential PI3K/Akt pathway activity. Post-NACT, stromal and vascular components underwent molecular and functional remodeling, shaping an immune-activated microenvironment. Mononuclear phagocytes resolved into three discrete clusters—monocytes, macrophages, and dendritic cells—with differentiation gradients. Endothelial cells maintained robust CXCL2 expression throughout the therapeutic course. Functional validation via transwell assays and Western blotting confirmed that endothelial-derived CXCL2 promoted macrophage chemotaxis via CXCR2, with corresponding CXCR2 upregulation in macrophages. Conclusions: Collectively, these findings suggest the complex cellular and transcriptional heterogeneity of the OS microenvironment and its chemokine-driven molecular remodeling after NACT, indicating that TME dynamics may be a determinant of therapeutic response and chemoresistance. Full article

Hemophagocytic lymphohistiocytosis (HLH) is associated with high mortality, underscoring the importance of rapid diagnosis and treatment. However, accurate and timely diagnosis is challenging due to its similarities with other acute inflammatory conditions. Traditional diagnostic frameworks rely on fever, lymphadenopathy, cytopenias, hyperferritinemia, hypofibrinogenemia, and related laboratory abnormalities, many of which may reflect either HLH or the underlying condition that triggered or mimics it. Newer diagnostics have emerged that are based on the underlying disease biology, particularly interferon-gamma (IFN-γ)-driven immune activation. C-X-C motif chemokine ligand 9 (CXCL9), a downstream marker of IFN-γ activity, has emerged as a promising adjunctive biomarker for HLH diagnosis, risk stratification, monitoring treatment response, and detection of disease reactivation. This narrative review examines the current evidence surrounding CXCL9 and its role in HLH. Full article

Background: Peripheral nerve injury drives persistent remodeling of the dorsal root ganglion (DRG) microenvironment, but the cellular states and intercellular signaling mechanisms that sustain neuropathic pain remain poorly understood. Methods: Here, we integrated single-cell transcriptomics with histological, behavioral, and electrophysiological approaches to identify an interferon-responsive state in satellite glial cells (SGCs) of the DRG following spared nerve injury (SNI). Results: This state was distinguished by STAT1-associated interferon-responsive genes’ activation and enhanced chemokine signaling, particularly involving CXCL10 and its receptor CXCR3. These molecular alterations were accompanied by remodeling of intercellular communication among SGCs, sensory neurons, and immune cells. Pharmacological inhibition of STAT1 or blockade of CXCR3 not only reduced sensory-neuron hyperexcitability but also attenuated pain hypersensitivity and improved deficits in hindlimb weight bearing and gait after SNI. Conclusions: Collectively, our findings indicate that interferon-responsive SGCs (IFN SGCs) contribute to the maintenance of neuropathic pain by regulating STAT1-dependent chemokine signaling in the DRG. Targeting this signaling pathway may provide a therapeutic strategy for persistent neuropathic pain. Full article

Topical non-steroidal anti-inflammatory drugs (NSAIDs) such as flurbiprofen and benzydamine may be used for the symptomatic relief of pharyngitis, triggered by infections. The onset and spectrum of anti-inflammatory activity of both drugs were compared in human tonsil (HTEpiC) and bronchial epithelial (BEAS-2B) cells stimulated with poly I:C against key inflammatory mediators. Cells were stimulated with poly I:C for 24 h and treated with flurbiprofen or benzydamine for 20 s, 2 min and 5 min. Production of prostaglandin E2 (PGE₂), tumour necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-1 beta (IL-1β), interleukin-8 (IL-8), interleukin-18 (IL-18), monocyte chemotactic protein-3 (MCP-3) and C-X-C motif chemokine ligand 10 (CXCL10), as well as cyclooxygenase-2 (COX-2) protein expression, was evaluated. Flurbiprofen significantly (p < 0.05) reduced PGE₂ production and COX-2 expression in both cell types within 20 s of treatment and maintained these effects at 2 and 5 min. However, benzydamine produced delayed, inconsistent inhibition. Although both drugs reduced poly I:C induced IL-6, IL-1β, IL-8 and IL-18 production, the magnitude and onset were greater with flurbiprofen. Flurbiprofen also reduced MCP-3 and CXCL10 in both cell types, alongside significant inhibition of caspase-1 activity. Benzydamine showed limited or no effect and less inhibition of caspase-1. With rapid suppression of the COX-2/PGE₂ pathway, rapid onset and a broad inhibition spectrum, flurbiprofen more comprehensively modulates inflammatory processes in epithelial respiratory models. Full article

Background/Objectives: Excessive inflammation is a critical contributor to impaired oral mucosal wound healing, yet effective therapeutic strategies are still lacking. Although dehydrocorydaline (DHC) has been reported to exhibit anti-inflammatory and analgesic properties, its role in wound healing and the underlying mechanisms have not been fully elucidated. This study aimed to investigate whether DHC accelerates palatal wound healing and to elucidate the role of the p38 mitogen-activated protein kinase (MAPK)/CCL2 signaling axis in DHC-mediated regulation of macrophage chemotaxis. Methods:In vitro, macrophages were stimulated with 1 μg/mL lipopolysaccharide (LPS) and treated with DHC at 0.1, 1, and 10 μM. The chemotactic response and inflammatory function of macrophages were assessed using real-time PCR, ELISA, Western blotting, and Transwell assays. Molecular docking simulations and Western blotting analyses were performed to examine the regulatory effect of DHC on MAPK signaling pathway. In vivo, a full-thickness palatal mucoperiosteal wound extending from the left maxillary first to third molars was established in mice by scalpel scraping. The effects of topical 10 μM DHC gel on wound healing were evaluated using stereomicroscopy, histological staining, and real-time PCR at 0, 3, and 5 days post-modeling. Results:In vitro, DHC effectively downregulated the expression of chemokines, including C-C motif chemokine ligand 2 (Ccl2), Ccl5, Ccl22, C-X-C motif chemokine ligand 10 (Cxcl10), and Ccl24, with the most significant inhibitory effect on Ccl2 (70.8% inhibition). In Transwell assays, DHC reduced macrophage migration by 68.2%. Mechanistically, DHC prominently inhibited the activation of the MAPK signaling pathway. In vivo, DHC treatment accelerated wound healing and markedly reduced macrophage infiltration in mouse palatal wound tissues. Conclusions: These findings demonstrated that DHC accelerated palatal wound healing 1.6-fold in mice. DHC suppressed macrophage chemotaxis by 68.2% through modulation of the MAPK signaling pathway. Full article

Periodontitis is a chronic dysbiotic inflammatory disease increasingly recognized as a contributor to systemic inflammatory burden and cardiovascular risk. Consensus and mechanistic literature support an association between periodontitis and atherosclerotic cardiovascular disease, while recent interventional evidence suggests that intensive periodontal treatment can improve endothelial function, reduce systemic inflammation and oxidative stress, and favorably influence the progression of carotid intima-media thickness. Among candidate mediators, interleukin-6 (IL-6) is especially compelling because it participates in local periodontal inflammation, hepatic acute-phase activation, endothelial dysfunction, oxidative stress, and the biology of atherosclerotic cardiovascular disease. CXCL10 has emerged as a relevant adjunctive chemokine because it is detectable in saliva, serum, gingival crevicular fluid, and inflamed periodontal tissues. In addition, it has also been implicated in atherosclerosis and adverse cardiac remodeling. Proteogenomic evidence suggests that CXCL10 may act as a potential downstream mediator of IL-6-associated signaling in atherosclerosis. Haptoglobin 2-2 (Hp2-2), while not itself a primary cytokine driver, may function as a host-susceptibility modifier, as the Hp2-2 phenotype has been associated with impaired hemoglobin scavenging, HDL dysfunction, endothelial dysfunction, oxidative vulnerability, and higher cardiovascular risk, particularly in dysglycemic states. This review proposes an IL-6-centered model linking chronic periodontal inflammation to cardiovascular risk, with CXCL10 positioned as an inflammatory recruitment amplifier and Hp2-2 as a genotype-defined modifier of vascular susceptibility. The translational implications of this framework for precision oral–systemic risk assessment, integration of salivary and blood-based biomarkers, and future biomarker-guided intervention studies are discussed. Full article

Background: The efficacy of chemotherapy in triple-negative breast cancer (TNBC) is limited by intrinsic resistance and the tumor microenvironment (TME). Accumulating evidence reveals a mechanistic connection between programmed death-ligand 1 (PD-L1) and c-x-c motif chemokine receptor 4 (CXCR4), which dominate stroma barriers, immune escape, and cancer metastasis. Earlier studies have shown that dual suppression of c-x-c motif ligand 12 (CXCL12)/CXCR4 and programmed cell death-1 (PD-1)/PD-L1 pathways regulates extracellular matrix (ECM) deposition, activation of cancer-associated fibroblasts (CAFs), and epithelial–mesenchymal transition (EMT) of pancreatic cancer cells. Methods: We combined BsNb PX4, a bispecific nanobody targeting PD-L1 and CXCR4, with paclitaxel or gemcitabine in multiple tumor cell lines and human peripheral blood mononuclear cell (hPBMC)-reconstituted xenograft mouse models. Antitumor activity was assessed by CCK-8, flow cytometry, and ELISA, and immune cell infiltration and TME remodeling were examined by immunofluorescence, immunohistochemistry, cytokine assays, and RNA-seq. Results: In MDA-MB-231 cells, BsNb PX4 synergistically enhanced paclitaxel-induced growth inhibition and apoptosis via G₂/M cycle arrest. This combinatorial strategy profoundly remodeled tumor immunity by expanding CD8⁺ T cells and depleting Foxp3⁺ CD4⁺ regulatory T cells (Tregs), while concurrently restoring T-cell cytotoxicity and skewing the cytokine balance toward an antitumor state, with elevated IFN-γ and reduced TGF-β1. Notably, compared with paclitaxel monotherapy, the combination significantly elevated intratumoral CD8⁺ T-cell infiltration, decreased Treg abundance, and exerted robust inhibitory effects on tumor growth and metastasis in humanized TNBC xenografts. Conclusions: These findings reveal that dual blockade of PD-L1 and CXCR4 acts synergistically with chemotherapy by triggering tumor cell apoptotic effects and reversing the immunosuppressive microenvironment, thereby emerging as a promising therapeutic strategy for TNBC. Full article

Vernal keratoconjunctivitis (VKC) is a chronic pediatric ocular allergy that can progress from seasonal to persistent inflammation with vision-threatening complications. Although Th2-associated mechanisms have been implicated, the immune correlates of disease severity are not well defined. Tear samples from VKC patients (moderate intermittent and moderate persistent) and healthy controls were collected using Schirmer’s strips. Cytokine profiling was performed using the OLINK^® Target 48 Cytokine Panel. Differential expression, correlation with clinical features, and pathway enrichment analyses were performed. Compared to control, IL-15, CXCL11, CXCL9, MMP12, and CCL13 were significantly elevated in VKC, with higher levels in the persistent phenotype. These cytokines correlated with symptom duration, limbal involvement, and papillary hypertrophy. Pathway analysis revealed enrichment of IL-17, JAK–STAT, and chemokine signaling pathways. VKC severity is associated with distinct tear cytokine signatures, with the persistent phenotype showing enhanced chronic inflammatory signaling. These findings identify candidate tear-based markers of disease severity that require validation in larger, independent, and longitudinal cohorts. Full article

Nucleated erythroid cells (NECs) are emerging as important immunoregulators at the feto–maternal interface, yet their chemokine profiles and functional dynamics across pregnancy remain poorly understood. Using a murine allogeneic pregnancy model (CBA × C57Bl/6), we isolated placental and splenic TER-119⁺ NECs at mid- (E12.5) and late (E19.5) gestation. Chemokine production (13-plex), chemokine receptor expression (qPCR), immunosuppressive molecules (PD-L1, TGF-β, and ROS), T-cell proliferation (CFSE), and immune cell migration (Transwell) were assessed. CD45⁺ placental NECs were the main producers of PD-L1, TGF-β, and ROS, with maximal expression at E19.5, suggesting their potential contribution to the immunosuppressive functions observed in the total TER-119⁺ population. Chemokine production showed a striking shift in CCL17 and CXCL9 from the spleen to the placenta as pregnancy advanced (E12.5 → E19.5). Splenic NECs displayed dominant expressions of CCR3 and CXCR4. Unexpectedly, CCL2 and CCL4 blockade enhanced immune cell migration toward placental NECs at E19.5. Placental nucleated erythroid cells potently suppressed T-cell proliferation at E12.5, and this suppressive capacity remained stable until full term. Placental NECs undergo dynamic chemokine reprogramming while maintaining stable T-cell suppression. The paradoxical enhancement of migration after CCL2/CCL4 blockade suggests a complex chemokine network warranting further investigation. These findings provide new insights into the immunobiology of pregnancy and may have implications for understanding pregnancy complications. Full article

Children with autism spectrum disorder (ASD) exhibit gut mucosal immune alterations, but the co-regulatory architecture linking stool immune proteins and cytokines within the same cohort remains unstudied. In 115 children (74 ASD, 41 controls; age 5–18 years), seven stool immune proteins (IgA subclasses, α₁-antitrypsin and calprotectin subunits) were quantified by UHPLC-MS/MS and ten by Luminex—chemokines eotaxin/CCL11 and IL-8/CXCL8 plus eight cytokines—each normalised to total protein. Age-adjusted partial Spearman correlations were computed for all 70 protein–cytokine pairs per stratum, using Benjamini–Hochberg correction, bootstrap confidence intervals and Fisher r-to-z tests. No pair survived FDR correction in any stratum. IgA1 and IL-1β/TP correlated positively across all strata (full cohort ρ = 0.409, 95% CI 0.131–0.634, n = 62; controls 0.583; ASD 0.210), with no significant between-group difference (Fisher z = 1.70, p = 0.090). Multiple imputation attenuated this to ρ = 0.265 (95% CI 0.055–0.452). An inverse trend between IL-1β/TP and CARS score (ρ = −0.336, n = 41) did not survive correction (p-FDR = 0.576). This power-limited, hypothesis-generating study identifies an exploratory IgA1–IL-1β mucosal axis present across groups, with no confirmed between-group difference. Adequately powered multi-centre studies are required. Full article

Chemokines are best known for orchestrating leukocyte migration; however, accumulating evidence indicates that mammalian CXCL14 also functions as a direct antimicrobial effector of innate immunity. Nevertheless, the molecular basis underlying the bactericidal activity of fish chemokines remains largely unexplored. In this study, we identified and functionally characterized Ctenopharyngodon idella CXCL14 (CiCXCL14) as a broad-spectrum antimicrobial protein. CiCXCL14 is predominantly expressed in immune-related organs and is significantly upregulated upon Aeromonas hydrophila challenge, suggesting an active role in antibacterial defense. Recombinant CiCXCL14 exhibited potent, concentration-dependent bactericidal activity against E. coli, A. hydrophila, P. fluorescens, A. veronii, S. aureus, and S. agalactiae. Time-kill assays further demonstrated that CiCXCL14 exhibited rapid bactericidal activity, achieving near-complete killing of S. aureus within 40 min and E. coli within 160 min. Mechanistically, CiCXCL14 disrupts bacterial membrane integrity and interacts with purified bacterial genomic DNA, indicating that membrane damage and DNA interaction may both contribute to its bactericidal activity. Notably, CiCXCL14 retained substantial antibacterial activity after heat treatment, whereas its activity gradually decreased with increasing NaCl concentrations. In vivo, administration of recombinant CiCXCL14 improved survival rates in grass carp challenged with a lethal dose of A. hydrophila and reduced bacterial burdens in the liver, spleen, and trunk kidney. Collectively, these findings identify CiCXCL14 as a teleost chemokine with direct antibacterial activity and demonstrate that it contributes to antibacterial defense through membrane disruption and interaction with bacterial DNA. These findings provide a foundation for future studies investigating the potential application of CiCXCL14 in aquaculture disease control. Full article

Background/Objectives: Ultra-high-magnification endoscopy (Endocyto) visualizes microscopic mucosal structures and has been proposed as a tool for assessing mucosal inflammation. We previously developed an endocytoscopic classification system (EC-A to EC-D) for grading the severity of inflammation. In this study, we investigated the relationship between endocytoscopic classification and microRNA (miRNA) expression in 36 patients with ulcerative colitis (UC). Methods: This cross-sectional study was conducted in two phases at two institutions. In the first phase, microarray analysis was performed on biopsy samples from patients with UC and healthy controls enrolled at Nagasaki University Hospital. In the second phase, 36 patients with UC who underwent total colonoscopy via Endocyto at Tottori University Hospital were included, and selected miRNAs were analyzed via quantitative polymerase chain reaction. Results: Differentially expressed miRNAs were identified, and four (miR-141-5p, miR-192-5p, miR-194-5p, and miR-215-5p) were downregulated in inflammatory areas. Notably, miR-192-5p expression was markedly lower in the EC-B and EC-C+D groups than in the EC-A group. Furthermore, expression of CXC motif ligand 2 (CXCL2), a pro-inflammatory chemokine, was upregulated in inflammatory tissues and negatively correlated with miR-192-5p expression. Conclusions: These results indicate that reduced miR-192-5p expression is inversely associated with CXCL2 expression and that, among the miRNAs examined, miR-192-5p downregulation most consistently corresponded to the endocytoscopic classification. Our findings suggest that Endocyto may serve as a real-time, in vivo adjunct for a histology-like assessment of mucosal inflammation; its clinical utility requires prospective validation. Full article