Search Results (2,185)

Background: Cystic fibrosis (CF) is an inherited multisystemic disease. Despite advances in treatment, many patients still experience progressive lung dysfunction. Dental follicle-derived mesenchymal stem cells (DF-MSCs) possess significant immunomodulatory potential in inflammatory airway diseases. We evaluated the effects of DF-MSCs on lymphocyte proliferation, CD4⁺CD25⁺FoxP3⁺ regulatory T cell (Treg) frequency, and cytokine responses in peripheral blood mononuclear cells (PBMCs) from CF patients. Methods: PBMCs from 20 CF patients and 20 matched healthy controls were isolated by density gradient centrifugation, stimulated with a CD3/CD28 T-cell-activating antibody cocktail (CD-mix), and co-cultured ex vivo with cryopreserved DF-MSCs. Lymphocyte proliferation was assessed by carboxyfluorescein succinimidyl ester (CFSE)-based flow cytometry, and Tregs were analyzed by flow cytometry. Cytokine levels in culture supernatants were quantified using a multiplex immunoassay. Results: DF-MSC co-culture significantly suppressed lymphocyte proliferation in CF PBMCs. Treg frequency significantly increased in the stimulated CF samples following MSC co-culture. Following co-culture, levels of tumor necrosis factor-alpha (TNF-α), interleukin-8 (IL-8), and IL-23 decreased. In CF samples, levels of interferon-alpha 2 (IFN-α2), monocyte chemotactic protein-1 (MCP-1), interleukin-12 (IL-12), interleukin-18 (IL-18), and interleukin-33 (IL-33) were elevated. Conclusions: DF-MSCs reduced lymphocyte proliferation, increased Treg frequency, and regulated cytokine levels in CF, supporting their potential to restore immune balance. Full article

Background: Progressive supranuclear palsy (PSP) is a primary 4-repeat tauopathy in which neurodegeneration may be accompanied by neuroinflammatory and peripheral immune alterations. Whether peripheral inflammatory activity reflects structural degeneration within vulnerable brain networks remains unclear. Methods: This retrospective case–control study included 12 patients with PSP and 12 patients with Parkinson’s disease (PD). Automated volumetric analysis of 3-Tesla MRI was performed using volBrain 2.0. Blood-derived inflammatory indices included neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), systemic inflammation response index (SIRI), and red blood cell distribution with coefficient of variation (RDW-CV). Results: Patients with PSP showed significantly lower normalized superior frontal gyrus and pallidal volumes than patients with PD. Within the PSP group, higher values of selected blood-derived inflammatory indices were associated with lower frontal network volumes. After adjustment for age, MLR was inversely associated with the composite Frontal Network Score (partial r = −0.7333, p = 0.0067, FDR q = 0.020). The strongest regional association was observed between SIRI and medial frontal cortex volume (rho = −0.748, p = 0.0051); however, regional associations did not remain significant after FDR correction. Conclusions: Peripheral inflammatory markers were associated with MRI-derived measures of frontal network degeneration in PSP. The association between MLR and the composite Frontal Network Score supports a link between systemic immune alterations and network-level neurodegeneration in PSP. Full article

Malaria pathogenesis involves complex immunopathological mechanisms that hinder early diagnosis and effective treatment. This study integrates multi-omics data and experimental models to identify host-derived biomarkers and elucidate their functional roles. By combining human transcriptomic datasets, weighted gene co-expression network analysis (WGCNA), and machine learning (LASSO, SVM, RF), we identified C1QB as a key hub gene. In human data, C1QB was significantly upregulated in both training and validation cohorts (AUC 0.983 and 0.970). Single-gene GSEA and immune infiltration analyses linked C1QB to apoptosis, inflammation, and altered immune cell composition, including increased activated dendritic cells and neutrophils, and decreased naïve B cells and CD8⁺ T cells. In a murine malaria model (Plasmodium berghei ANKA), C1QB expression rose as early as day one post-infection, preceding detectable parasitemia. Immunohistochemistry revealed C1QB accumulation in the liver and spleen. Single-cell RNA sequencing in the murine model confirmed monocyte-predominant expression, and scTenifoldKnk analysis suggested its role in immune regulation. Crucially, inhibiting C1q in mice via antibody intervention alleviated malaria-induced inflammation, tissue damage, and apoptosis, indicating that C1QB/C1q actively contributes to immunopathology. AI-based drug prediction and molecular docking further supported its therapeutic potential. Collectively, our findings establish C1QB as a dual biomarker and pathogenic driver in malaria, with diagnostic and therapeutic implications. Further studies are required to validate direct target engagement and clarify upstream regulatory mechanisms. Full article

Major airway diseases, including chronic obstructive pulmonary disease (COPD), asthma, bronchiectasis and chronic rhinosinusitis without nasal polyps (CRSsNP), frequently coexist and share inflammatory, epithelial and remodeling features. However, whether these clinically distinct airway disorders are driven by a unified genetic liability and how this shared liability maps to disease-relevant tissues, genes and immune-regulatory programs remain incompletely understood. We integrated GWAS summary statistics for COPD, asthma, bronchiectasis and CRSsNP using linkage disequilibrium score regression, local genetic correlation analysis and Genomic structural equation modeling. A latent shared airway disease factor, termed gAirwayDisease, was constructed to capture common genetic liability across the four conditions. We then applied an integrative functional genomics framework, including gsMap spatial enrichment, PoPS gene prioritization, MAGMA gene-set enrichment, GTEx v8 lung MTWAS, OneK1K and DICE immune-cell MTWAS, scMORE regulon analysis and phenome-wide Mendelian randomization. All six airway disease pairs showed positive genetic correlations, with estimates ranging from 0.508 to 0.685. Genomic SEM supported a single shared factor, with positive standardized loadings for COPD, asthma, bronchiectasis and CRSsNP and excellent model fit. Spatial mapping localized gAirwayDisease-associated signals to airway- and epithelial-associated anatomical domains. PoPS prioritized immune and airway-relevant genes, including SMAD3, GATA3, IL1R1, RUNX3 and STAT6, while MAGMA enrichment highlighted B-cell activation, T-cell activation and transcriptional regulatory pathways. Lung MTWAS identified SLC9A2 and ORMDL3 as top genetically regulated expression signals. OneK1K immune-cell MTWAS highlighted recurrent IL18R1 associations across CD4 and CD8 T-cell subsets. scMORE further identified 36 significant regulon–cell type pairs across dendritic cells, B cells, monocytes, T cells and NK cells, including BCL11A, TCF4, KLF4, RUNX1 and STAT4 regulons. MR-PheWAS linked genetically predicted gAirwayDisease to respiratory, allergic, lung function and immune-related traits. This study defines gAirwayDisease as a genetically informed latent factor capturing shared liability across major airway diseases. Integrated functional genomic analyses highlight airway epithelial and immune regulatory programs associated with shared disease susceptibility and prioritize candidate genes and regulons for future experimental validation. Full article

Bone-resident macrophages are increasingly recognized as regulators of bone metabolism, yet their contribution to primary human osteoblast cultures remains poorly understood. We investigated the functional relevance of CD14-positive cells in primary human osteoblast cultures obtained using two isolation methods. CD14-positive cells were depleted by magnetic-activated cell sorting, and non-depleted and depleted cultures were compared regarding cell viability, matrix mineralization, osteogenic and macrophage-associated marker expression, and responses to IL-1β, IL-6, and TNF-α. CD14 depletion reduced cell viability under growth conditions and significantly impaired matrix mineralization in cultures obtained with both isolation protocols. Depletion markedly decreased CD14 and other monocyte/macrophage-associated markers, while the osteogenic cell population was largely preserved, although marker-specific changes suggested altered osteoblast maturation. Despite reduced basal mineralization, IL-1β and TNF-α significantly enhanced mineralization in both non-depleted and CD14-depleted cultures, whereas IL-6 had no significant effect. These findings indicate that CD14-positive cells support basal osteoblast viability, maturation, and mineralization but are not required for the mineralization-promoting effects of IL-1β and TNF-α. Primary human osteoblast cultures should therefore be considered multicellular systems in which macrophage-like cells contribute substantially to basal osteogenic function. Full article

Exosomes carry bioactive macromolecules driving sepsis pathogenesis, but the mechanisms underlying macrophage-to-hepatocyte communication during systemic inflammation remain poorly understood. We investigated how sepsis-induced macrophage exosomes are involved in remote intercellular crosstalk with hepatic cells via transcription factor-encoding mRNA cargo. Human monocytic THP-1 macrophages were stimulated with lipopolysaccharide (LPS), followed by exosome isolation, recipient cell uptake verification, and high-throughput RNA sequencing cargo analysis. To functionally reconstruct downstream signaling in recipient cells, exosome-enriched MAFB mRNA was transiently overexpressed in a HepG2 cell model, with subsequent expression changes mapped at both the transcript and protein levels using quantitative PCR and Western blot analyses. This ectopic MAFB expression directly upregulates the expression of microRNA-155 (miR-155). Crucially, elevated miR-155 acts as a post-transcriptional repressor that directly targets and downregulates JARID2 and H3F3A mRNAs and their corresponding protein products within the liver cells, orchestrating a “repressor-of-repressors” disinhibition cascade that drives net chromatin remodeling and activation of downstream hepatic target genes. This study demonstrates that sepsis alters exosomal transcription factor mRNA cargo and delineates a mechanistic downstream pathway—MAFB → ↑miR155 → ↓Jarid2 & ↓H3F3A → Chromatin Remodeling → Downstream Hepatic Gene Activation pathway—that provides novel, specific molecular checkpoints for therapeutic intervention in sepsis-induced liver injury. Full article

The diagnosis and clinical management of myelodysplastic neoplasms are increasingly challenged by the disease’s inherent heterogeneity, particularly with respect to the diagnosis of low-grade variants. While standardized flow cytometric protocols traditionally rely on static biomarkers for lineage assignment, these often fail to capture the dynamic biological behavior of the malignant clone. This review synthesizes studies on the integration of functional biomarkers, specifically the nuclear proliferation marker Ki-67 and the anti-apoptotic protein Bcl-2, into the diagnostic and prognostic workflow. By utilizing high-dimensional multiparameter flow cytometry (MFC) and software-based maturation continuum analysis, the survival and growth kinetics of the myeloid, erythroid, and monocytic lineages can be quantified. These findings redefine myelodysplastic syndromes (MDS) as characterized by a significant decrease in cell-cycle progression and an increase in anti-apoptotic activity during early stages of maturation. Recent studies demonstrate that integrating the erythroid Ki-67 proliferation index as a fifth parameter into the conventional Ogata score dramatically improves diagnostic sensitivity for detecting MDS from 66% to 90% while maintaining 100% specificity. In particular, the sensitivity for detecting low-grade MDS improved from 56% to 91%. Additionally, a reduced erythroid Ki-67 index (≤28%) is a powerful independent predictor of transfusion dependence within 1 year. Beyond diagnostics, the introduction of the Bcl-2:Ki-67 ratio provides a superior metric for biological aggressiveness and a potential predictive tool for precision medicine. A high ratio identifies a quiescent, apoptosis-resistant cell population that is likely refractory to standard chemotherapy but is an ideal candidate for targeted Bcl-2 inhibition with Venetoclax. The integration of functional biomarkers bridges the gap between complex mutational landscapes and clinical manifestations. While digital imaging and artificial intelligence (AI) are beginning to automate blast enumeration and maturation analysis, functional kinetics may provide a necessary biological readout for personalized therapy. Full article

Atherosclerosis remains the principal underlying cause of myocardial infarction, ischemic stroke and peripheral artery disease, and its progression reflects a complex interplay between lipid accumulation, endothelial dysfunction, chronic vascular inflammation and maladaptive remodeling of the arterial wall. The Notch signaling pathway, an evolutionarily conserved juxtacrine communication system, has emerged as a central regulator of every cell type implicated in atherogenesis, including endothelial cells, vascular smooth muscle cells, monocytes/macrophages and T lymphocytes. Depending on the receptor–ligand pairing, the hemodynamic context and the cellular compartment involved, Notch signaling can be either atheroprotective or atherogenic, a duality that has complicated efforts to translate mechanistic insight into therapy. This review summarizes current knowledge of the molecular architecture of the Notch pathway in the vasculature, dissects its cell type-specific and stage-specific contributions to atherosclerotic plaque initiation, progression, calcification and destabilization, and critically appraises pharmacological strategies designed to modulate Notch activity, including γ-secretase inhibitors, ligand- and receptor-directed monoclonal antibodies, soluble decoy receptors, microRNA-based approaches and drug repurposing strategies such as statins. Particular attention is paid to the cardiovascular toxicities that have emerged from oncology trials of Notch pathway inhibitors, which illustrate both the pharmacological tractability and the narrow therapeutic window of this pathway. We conclude that Notch-directed therapy for atherosclerosis is mechanistically well justified but will require cell type-selective and context-selective delivery strategies to be clinically viable. Full article

Vaccinium macrocarpon (American Cranberry) fruit is processed to make juice, supplements, and other edible products, leaving pomace as a sidestream that contains secondary metabolites with potential anti-inflammatory and anti-proliferative activities. Ultrasound-assisted extraction and chromatographic methods were developed to prepare an extract (POM-ACE) and fractions rich in pentacyclic triterpenoids and phytosterols for bioactivity evaluation. These were characterized using UPLC-MS and GC-MS. Anti-inflammatory effects were assessed in a human monocyte (THP-1) model, identifying several fractions containing triterpenoids and sitosterol that significantly inhibited IL-1β expression. Molecular docking indicated favorable interactions of these components with NLRP3, suggesting a possible modulation of inflammasome. Fractions rich in ursolic acid (UA) and oleanolic acid (OA) and their p-hydroxycinnamic acid (HCA) esters also showed mild concentration-dependent inhibition of lipoxygenase (LOX) activity. Pearson correlation analysis identified strong positive correlations between UA, OA, and their trans-HCA esters with LOX inhibition. Anti-proliferative activity was assessed in HT-29 colon adenocarcinoma cells using MTT, with several fractions exhibiting moderate concentration-dependent activity (IC₅₀ = 12–17 µg/mL). PLS regression analysis supported significant contributions by UA, OA, and their HCA esters. Flow cytometry experiments demonstrated that cell death occurs in part through apoptosis; the role of apoptosis was further supported by favorable molecular docking interactions between pomace triterpenoids and caspases-3 and -9. These findings suggest that cranberry pomace is a promising source of triterpenoids and phytosterols with anti-inflammatory and anti-proliferative properties. Full article

Therapies based on mesenchymal stromal cells (MSCs) have high potential in the field of regenerative medicine due mainly to their immunomodulatory properties. However, their clinical translation is hampered by a lack of sufficiently standardised potency tests. Since macrophages constitute key mediators of the effects of MSCs, macrophage-based assays potentially provide a relevant in vitro tool for the evaluation of the activity of MSC products. This study involved the coculturing of canine adipose-derived mesenchymal stromal cells (ASCs) with macrophages derived from human THP-1 and U937 monocyte cell lines, murine RAW264.7 macrophages and primary human macrophages. The M2 polarisation was assessed following stimulation with IL-4/IL-13 in THP-1 and U937 macrophages. The mRNA expression of the pro- and anti-inflammatory markers was analysed using qPCR. The ASC transwell coculture altered the LPS-induced inflammatory mRNA expression in a strongly model- and marker-dependent manner. The U937-derived macrophages exhibited the most consistent suppression of the tested inflammatory transcripts and the RAW264.7 cells provided a practical readout for selected inflammatory markers, whereas the THP-1 macrophages evinced the suppression of TNFA but not IL1B or PTGS2 under the selected stimulation conditions. IL-4/IL-13 induced moderate but statistically non-significant changes in IL10 and TGFB1 in the U937-derived macrophages but no reproducible response in the THP-1-derived macrophages. In a subsequent U937 coculture experiment, ASC-derived paracrine factors altered selected M2-associated transcripts at specific time points. The results thus provided support for macrophage-based transcriptional readouts as an early-stage tool for comparing responder macrophage models and detecting the selected anti-inflammatory paracrine effects of canine ASCs; the U937 cells were found to be particularly suitable for the study of inflammatory polarisation and the RAW264.7 cells for the purpose of standardised screening. Full article

C-C motif chemokine ligand 2 (CCL2) interacts with cytokines, adipokines, miRNAs, and multiple synovial cell populations. Experimental studies indicate that these interactions can form a CCL2-associated inflammatory amplification network across cell types. In cellular and animal models, increased CCL2 is associated with monocyte recruitment, synovial fibroblast activation, osteoclast-related bone remodelling, and vascular responses. Therapeutic strategies targeting the CCL2-centered inflammatory network include antagonists of the CCL2/CCR2 axis, natural products, synthetic compounds, conventional antirheumatic drugs, and emerging delivery-based approaches. Notably, direct CCL2/CCR2 inhibition has shown biological activity in experimental models but has not produced consistent clinical benefit in established rheumatoid arthritis (RA). Although these findings do not establish CCL2 as a dominant causal driver of RA, human observational studies suggest that circulating CCL2 may complement established markers in preclinical RA risk assessment, disease activity and remission classification, estimation of treatment response, and evaluation of RA-related complications such as interstitial lung disease. Of note, no validated concentration cut-off or standardized assay currently supports its routine clinical use. This review examines the CCL2-related inflammatory network in RA and evaluates its cellular mechanisms, therapeutic implications, and potential clinical applications. Full article

Primary membranous nephropathy (pMN) is an antibody-mediated podocytopathy, most commonly caused by autoantibodies against the M-type phospholipase A2 receptor (PLA2R1). Rituximab (RTX), an anti-CD20 monoclonal antibody, is a first-line treatment for moderate-to-high-risk pMN, inducing partial or complete remission in about 60% of patients within 24 months. However, treatment response varies considerably, and current biomarkers, including anti-PLA2R1 titers and peripheral B cell counts, have limited predictive value for non-response or relapse. Beyond B cell depletion, RTX exerts broader immunomodulatory effects by influencing T cell subsets, monocytes, and natural killer (NK) cells involved in antibody-dependent cellular cytotoxicity. This review examines the peripheral immune cell changes that accompany anti-CD20 therapy and their value as candidate biomarkers. Total CD19+ B cell depletion is the standard pharmacodynamic measure of drug effect but correlates only loosely with clinical outcome. A specific B cell reconstitution profile was associated with pending relapse. Class-switched memory B cells remain depleted during sustained remission, and their premature re-expansion has been associated with subsequent relapse. Regulatory T cells are reduced in active disease and rise within days of infusion in patients who later respond. The systemic inflammation response index, derived from the routine differential count, has been associated with both 6- and 12-month remission. These observations derive from small, mostly single-center cohorts using heterogeneous panels and different RTX regimens. On the available evidence, immune cell profiling cannot yet be recommended for routine disease monitoring, and larger prospective studies with standardized panels are required. Full article

Background: One of the key hallmarks of aging is the age-related decline in immune system function, accompanied by a chronic low-grade inflammation, or “inflammaging”. Simultaneously, a reduced capacity of immune cells to recognize and eliminate pathogens, along with immune exhaustion, is also defined as a sign of aging. Cynomolgus macaques (Macaca fascicularis) belong to a group of non-human primates evolutionarily close to humans and are often used for preclinical research. Methods: In this study, we performed mRNA sequencing of bone marrow and peripheral blood samples from young (5 years old) and old (over 19–21 years old) cynomolgus macaques to identify key markers of immunosenescence. Results: Although an increase in p16 expression was detected, we did not observe the increase in the senescence-associated secretory phenotype (SASP) cytokines reported in previous studies. Instead, we observed a transcriptional profile characterized by increased lymphocyte cytotoxic activity combined with a decrease in proinflammatory signaling, reduced markers of myeloid cells, and lowered sensitivity to pathogen-associated patterns. Similar changes were detected in both blood and bone marrow: decreased expression of naive T-cell markers (CCR7, LEF1, SELL, and FOXO1), reduced markers of the myeloid lineage—neutrophils and monocytes (CD177, CD14, CD163, FPR1, FPR2, and CXCR1)—and downregulation of genes belonging to different pattern-recognition receptor families (TLR1, TLR2, TLR4, TLR5, TLR6, TLR8, TLR10, IFIH1, CLEC4E, NOD2, NLRC4, NLRP12, NLRX1, and NAIP). In contrast, the group of old animals showed increased expression of markers associated with terminally differentiated cytotoxic lymphocytes (CD8+ T cells and NK cells): GZMB, PRF1, KLRK1, FASLG, TBX21, CCR5, and GNLY. Conclusions: Our findings offer new perspectives on the molecular mechanisms of age-associated immune dysregulation in non-human primates, serving as a baseline for selecting key candidate genes in subsequent functional investigations. Full article

Indolo[2,3-b]quinoxalines, along with their N-substituted derivatives, exhibit pronounced anticancer activity, although the mechanisms of their biological action may vary. Herein, a panel of sixty-five 6-(2-aminoethyl)-6H-indolo[2,3-b]quinoxaline derivatives comprising eight series with distinct amine moieties connected to the tetracyclic indoloquinoxaline core via a dimethylene linker was evaluated as drug-like candidates for kinase binding and cytotoxic activity. The ADME (Absorption, Distribution, Metabolism, and Excretion) properties of the compounds included in this set were preliminarily determined using the SwissADME tool. Analysis revealed that the library of quinoxaline derivatives largely complies with the drug-likeness rule for kinase-targeted compounds. As part of the biological screening, the compounds were initially tested on two cell lines MonoMac-6 and THP-1 (both derived from patients with acute monocytic leukemia) using sunitinib, a known antitumor agent acting as a multi-target receptor tyrosine kinase inhibitor, as a reference compound. Compound 3g, which demonstrated the highest activity in the cytotoxicity analysis (IC₅₀ = 1.9 and 3.5 μM for the MonoMac-6 and THP-1 cell lines, respectively), was screened using the Eurofins DiscoverX scanEDGE panel, comprising 97 distinct kinases representing all known kinase families. Subsequently, the compound was tested using the Eurofins DiscoverX scanTK™ panel, covering 135 distinct receptor and non-receptor tyrosine kinases. Based on initial screening results, compound 3g exhibits relatively high binding activity against fourteen tyrosine kinases, including TYK2, ZAP70, eight mutant forms of ABL1, two mutant forms of FLT3, and one mutant form of ALK, and demonstrates relatively high binding selectivity with respect to non-mutant tyrosine kinases (S-score: 0.024). Secondary screening of nine selected analogs of compound 3g led to the identification of compound 3h, which demonstrates relatively high binding affinity for FLT3 (D835V) (K_d = 0.41 μM). Molecular modeling suggested modes of binding interaction of the compounds 3h and 3g in the FLT3 (D835V) catalytic site. Our results demonstrate that 6-(2-aminoethyl)-6H-indolo[2,3-b]quinoxaline derivatives could be potential candidates for developing anticancer drugs. Full article

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is traditionally known for regulating plasma LDL cholesterol levels via LDL receptor degradation. This exploratory study examined the association between chemically induced changes in Krüppel-like factor 2 (KLF2), a vasoprotective transcription factor, and PCSK9 expression under ox-LDL-induced conditions in RAW264.7 cells. In silico molecular docking was also performed to determine whether GGTI298 could adopt a predicted binding pose within PCSK9. KLF2 was modulated by adding GGPP (a chemical inhibitor) and GGTI298 (a chemical activator) to the monocytes. Quantitative real-time PCR and immunocytochemistry were used to assess KLF2 and PCSK9 expression. Molecular docking was performed to examine the interaction between GGTI298 and PCSK9 using computational tools. Results show that ox-LDL significantly increased PCSK9 expression in monocytes during foam cell formation. However, GGPP significantly increased PCSK9 expression. In contrast, GGTI298 markedly reduced PCSK9 expression, suggesting a negative regulatory role of KLF2. Docking studies revealed that GGTI298 binds to the PCSK9 catalytic domain with favorable binding energy, forming stable hydrogen bonds and hydrophobic interactions with key amino acid residues, indicating potential interference with PCSK9 function. GGTI298 suppresses PCSK9 expression during foam cell formation, highlighting its protective role. The dual ability of GGTI298 to enhance KLF2 expression and directly bind to PCSK9 underscores its potential as a therapeutic agent for managing foam cell formation that leads to atherosclerosis. The findings demonstrate an inverse association between KLF2 and PCSK9 expression following chemical treatment under ox-LDL-induced conditions. However, direct KLF2-dependent regulation of PCSK9, direct GGTI298–PCSK9 binding, and functional inhibition of PCSK9 were not established and require further genetic, biochemical, and functional validation. Full article