INTRODUCTION:HIV screening by chemiluminescent immunoassay (CLIA) enables the simultaneous detection of anti-human immunodeficiency virus (HIV)-1/2 antibodies (Abs) and the p24 antigen (Ag), ensuring high sensitivity and specificity. Viral typing is essential for the clinical follow-up of seropositive blood donors. This study aimed to identify anti-HIV Ab profiles among blood donors who tested positive by CLIA and to assess the concordance between CLIA screening and supplemental rapid testing. METHODS:A descriptive study was conducted at the Regional Blood Transfusion Center of Bouaké from November 2024 to February 2026. Blood donors were screened for HIV using two CLIA platforms (MAGLUMI and MACCURA). The testing algorithm consisted of CLIA screening, followed by HIV Bioline rapid testing for serological typing and a follow-up test on a new blood sample. CLIA-reactive samples were classified as concordant reactive when confirmed by the HIV Bioline test and as presumed false positives when the initial CLIA-reactive result was not confirmed by HIV Bioline and became non-reactive on repeat CLIA testing on a new sample. RESULTS:Among 21,643 blood donors screened, 79 (0.37%) were reactive by CLIA. A male predominance was observed (sex ratio: 2.8). The mean age was 33.9 years in men and 31 years in women. Of these, 44/79 (55.7%) were classified as concordant reactive, 19/79 (24.1%) as presumed false positives, and 16/79 (20.2%) as indeterminate (CLIA-reactive but HIV Bioline-negative pending follow-up). Among the confirmed reactive samples, 34 (43.0%) were infected with HIV-1, 1 (1.3%) with HIV-2, and 9 (11.4%) had HIV-1/HIV-2 dual infection. Two cases of hepatitis B co-infection were also identified. The MAGLUMI assay showed a specificity of 99.93%, a proportion of concordant reactive results of 77.5%, and a presumed false-positive rate of 0.07%, whereas the MACCURA assay showed a specificity of 99.87%, a concordance of 65%, and a presumed false-positive rate of 0.13%. Signal-to-cutoff (S/CO) ratios were higher in concordant reactive samples (1.89-500) than in presumed false-positive samples (1.03-11.56). CONCLUSION:Despite their high performance, CLIA assays require confirmatory testing to determine the HIV type (HIV-1 or HIV-2), reduce false-positive results, and ensure a reliable diagnosis.
Asthma and chronic obstructive pulmonary disease (COPD) are chronic respiratory diseases associated with inflammation and structural changes in the airways. Hypoxia is a key element of the pulmonary microenvironment, particularly in obstructive diseases, where it can be transient (e.g., during asthma exacerbations) or chronic (COPD). Pulmonary vascular endothelial cells (ECs) are important components of the immune response and (like the epithelium) are exposed to hypoxia and viral infections. Although it has been demonstrated that ECs can be infected by respiratory viruses and that hypoxia can affect their function, the impact of hypoxia on the antiviral capabilities of the endothelium remains poorly understood. This study examined the influence of acute and chronic hypoxia on the antiviral properties of human lung microvascular ECs. Acute and chronic hypoxia upregulated endosomal RNA receptors TLR3 and TLR7, whereas they downregulated cytosolic RIG-I and MDA5. Acute hypoxia downregulated mRNA and protein expression of IRF3 and IRF7 and inhibited effector antiviral mechanisms, whereas chronic hypoxia induced IRF7, OAS-1, protein kinase-R (PKR), and Mx1 at both mRNA and protein levels by flow cytometry. In addition, both hypoxia models caused increased secretion of lymphocyte-recruiting chemokines, whereas the secretion of cytokines/chemokines responsible for the involvement of monocytes and macrophages was inhibited. Acute hypoxia suppresses antiviral regulatory and effector pathways, whereas chronic hypoxia enhances viral RNA detection and intracellular antiviral mechanisms. These findings highlight the complex role of hypoxia in shaping endothelial immune responses and suggest that, depending on its duration, it may influence susceptibility to viral infections in chronic respiratory diseases.
HIV-1 reservoirs are predominantly located in CD4+ T-cells; however, not all CD4+ T-cells contribute to the reservoir in the same way. Factors such as activation, differentiation, and cell metabolism have been proposed to determine the relative susceptibility of cells to HIV-1 infection. HIV-1 reservoirs are seeded early during the acute phase of infection, but the cell composition of these reservoirs evolves in the transition to chronic infection. This suggests that there are factors during the acute phase that may alter the intrinsic susceptibility of CD4+ T-cells to HIV-1. We investigated here the influence of common cytokines known to be secreted during the acute phase and to play a role in either T cell homeostasis or HIV-1 infection on activation, differentiation, metabolic activity, and HIV-1 susceptibility of CD4+ T-cells. We show that the proinflammatory cytokines interleukin (IL)-2, IL-7, and IL-15 induce cellular activation, differentiation to effector profile, increase in metabolic capacity, and HIV-1 susceptibility. In contrast, IL-21, another cytokine from the gamma-chain (γc) family, showed opposing effects on the same parameters, including decreasing oxidative phosphorylation (OXPHOS) and HIV-1 infection. We also show that IL-10 and interferon (IFN)-α are able to at least partially revert the cellular and metabolic changes induced by IL-2 or IL-7 and reduce the cells' susceptibility to HIV-1.
Oncology is undergoing a profound transformation driven by the convergence of artificial intelligence (AI), RNA-based vaccines, and chimeric antigen receptor T-cell (CAR-T) therapies. Individually, these technologies have advanced cancer diagnosis, treatment, and patient stratification. AI-driven approaches enhance drug discovery, optimize clinical trial design, and enable personalized therapeutic decision-making. RNA vaccines provide a flexible platform for encoding tumor-specific neoantigens, while CAR-T therapies enable targeted immune-mediated tumor cell elimination. Early-phase clinical trials, particularly those combining RNA vaccines with immune checkpoint inhibitors, have demonstrated promising improvements in recurrence-free survival (RFS) and immunogenicity. However, evidence supporting direct combinations of RNA vaccines and CAR-T therapies remains largely preclinical or limited to early-phase investigation and should therefore be interpreted with caution. This review explores how AI facilitates neoantigen discovery, RNA vaccine optimization, and CAR-T cell engineering, and examines the emerging interplay between these modalities. While their integration represents a compelling framework for personalized oncology, significant challenges remain, including clinical validation, scalability, regulatory oversight, and equitable access.
Acute graft-versus-host disease (aGvHD) remains a substantial cause of morbidity and mortality after allogeneic hematopoietic cell transplantation (allo-HCT). Limited studies explore the association between specific major histocompatibility complex (MHC) alleles and aGvHD or transplant-related mortality (TRM). After a noted clinical trend of higher TRM among a series of patients with aGvHD and MHC class I HLA- A ∗ 01:01, we retrospectively evaluated transplant outcomes of 404 adult allo-HCT patients who underwent unmodified allografts between 03/2010 and 02/2017. HLA- A ∗ 01:01 was expressed by 104 (25.7%) patients. In a univariate analysis, patients who underwent unmodified transplants and expressed HLA- A ∗ 01:01 had a higher risk of TRM (HR 1.63 [95% CI: 1.05-2.53], p = 0.035). In a multivariate cause-specific Cox model, HLA- A ∗ 01:01 was significantly associated with TRM after adjusting for grade II-IV aGvHD, conditioning regimen intensity, and age at transplant (HR 1.59 (95% CI: 1.02-2.48) p = 0.039). There were no significant differences in overall survival (OS), aGvHD, or chronic GvHD based on HLA- A ∗ 01:01 expression. With confirmation in a larger cohort, these findings have potential implications for the selection of allograft type, conditioning regimen, and post-transplant monitoring for this high-risk population.
Acute-on-chronic liver failure (ACLF) is a fatal syndrome defined by hepatic decompensation and systemic inflammation, yet the underlying cellular networks remain elusive. Here, we established an ACLF mouse model recapitulating clinical hallmarks and performed single-cell RNA sequencing (scRNA-seq) on hepatic non-parenchymal cells (NPCs). We identified 30 cell clusters and observed profound spatial remodeling of NPCs driven by pro-inflammatory mediators. Endothelial cells (ECs) underwent a critical transition from homeostasis to dysfunction, marked by mitochondrial damage and activation of NF-κB and MAPK pathways. Ligand-receptor interactome analysis identified dysfunctional ECs as central hubs driving global network reconfiguration, primarily via the Lgals9-Ighm/Cd45/Cd44 axes, alongside the suppression of homeostatic signals (Cd55 and APP). Furthermore, we mapped B cell lineage trajectories and validated the spatial co-localization of EC and B-cell interactions through Lgals9-Ighm. This study defines the EC and B-cell communication landscape and suggests that the Lgals9-Ighm axis may play a critical role in ACLF progression, representing a potential candidate for therapeutic intervention.
Toll-like receptors (TLRs) and P2 receptors are key regulators of innate immunity. During infection, pathogen-associated molecular patterns (PAMPs) activate TLRs, whereas extracellular nucleotides engage P2 receptors and influence inflammatory responses. Here, we show that polyinosinic:polycytidylic acid (poly[I:C])-activated TLR3 and P2Y2 signalling functionally interact in intestinal epithelial cells (IECs) to regulate chemokine production. HT-29 cells were stimulated with PAMPs, including the TLR3 agonist poly(I:C), in the presence or absence of P2 receptor signalling inhibitors. CXCL8/IL-8 or CXCL10/IP-10 secretion was assessed by ELISA and mRNA expression by quantitative real-time PCR (RT-qPCR). Primary IECs from P2Y2 knock-out (KO) and wild-type (WT) mice were also treated with poly(I:C), and CXCL1/KC secretion was measured. Poly(I:C) stimulation induced robust CXCL8/IL-8 and CXCL10/IP-10 release in HT-29 cells. This response was inhibited by nucleotide scavenging, P2 receptor blockade, and P2Y2 targeting using either specific antagonists or siRNAs. P2Y2 was the dominant receptor expressed, and its ligands ATP/UTP constitutively released by these cells amplified CXCL8/IL-8 production induced by a suboptimal concentration of poly(I:C), while alone they had no effect. In support of these findings, primary IECs from P2Y2 KO mice secreted significantly less CXCL1/KC than WT controls. Altogether, extracellular nucleotide signalling regulates TLR3-induced chemokine release in IECs through P2Y2 receptors.
Staphylococcus aureus remains a major cause of hospital- and community-acquired infections, successfully evading host defense mechanisms and escaping proposed vaccination strategies. The effective response of professional phagocytes is crucial in limiting the spread, tissue invasion, and subsequent infection by S. aureus. However, direct stimulation of innate immunity with S. aureus antigens is too risky due to their strong proinflammatory properties. Therefore, this study aimed to test the hypothesis that plant extracts trigger the gentle release of staphylococcal bioactive components that can modulate innate immunity. This strategy was tested in vitro; however, it may have future applications by employing the resident microbiota and dietary supplements containing plant extracts. In this study, the THP-1-derived macrophages were exposed to the supernatants of planktonic and biofilm S. aureus cultures pretreated with Viburnum opulus L. bark and fruit extracts. Vancomycin (VAN) and chlorogenic acid (ChA), used at subinhibitory concentrations, were included for comparison regarding the release of S. aureus active components. The cells were also exposed to purified bacterial cell wall components. Various elements of the immune response were assessed, including surface expression of cluster of differentiation (CD11c, CD206) via flow cytometry, cytokine production (tumor necrosis factor-alpha [TNF-α] and interleukin 10 [IL-10]) via ELISA, and S. aureus phagocytosis and intracellular killing using fluorescein isothiocyanate (FITC)-labeled bacteria and Alamar Blue staining. It was demonstrated that both supernatants from planktonic and biofilm S. aureus cultures pre-exposed to V. opulus extracts, as well as the bacterial cell wall components themselves, significantly increased TNF-α and IL-10 production by THP-1-derived macrophages and visibly reduced intracellular multiplication of engulfed S. aureus. The lack of changes in the surface CD marker expression may be a limitation of the model used (phorbol-12-myristate-13-acetate [PMA]-differentiated THP-1) rather than the phenomenon itself, that leads to a "ceiling effect". To sum up, the study presents V. opulus extracts as the triggers to release bioactive components from S. aureus cells, able to target innate immunity against staphylococci, which may be used in the future.
Zika virus (ZIKV) infection represents a critical threat to maternal-fetal health due to its ability to cross the placental barrier, converting this normally protective organ into a direct target of viral infection. This study examines the mechanisms by which ZIKV infects placental cells, including trophoblasts and Hofbauer macrophages, through the engagement of specific cellular receptors such as AXL and T-cell immunoglobulin and mucin (TIM), while simultaneously evading host immune defenses. Although the placental immune system employs interferon signaling and innate immune cells to restrict viral replication, ZIKV disrupts these pathways, promoting chronic inflammation and structural damage within placental tissue. These morphofunctional alterations impair fetal nutrient exchange and facilitate vertical transmission, leading to severe adverse outcomes such as congenital Zika syndrome (CZS), intrauterine growth restriction, and preterm birth. Importantly, pregnancy outcomes appear to depend on a delicate balance between the timing of maternal infection and the effectiveness of the local immune response. Finally, this study highlights the need for advanced molecular approaches and "omics" technologies to address existing knowledge gaps regarding viral variability and to inform the development of future therapeutic strategies aimed at preventing fetal transmission.
Enteroviruses (EVs) lead to 10-15 million symptomatic infections each year in the United States alone. EVs enter through either the gastrointestinal or respiratory mucosa, depending on the specific virus. Since these mucosal surfaces are the primary entry points for viruses, mucosal immunity serves as a crucial first line of defense. Nevertheless, the mechanisms that drive it are poorly understood for most EVs. Conventional intramuscular vaccines provide limited protection at these sites, leaving them unprotected; mucosal vaccination addresses this issue directly by generating local SIgA in GALT/NALT that neutralizes pathogens before they spread systemically, while also offering advantages in accessibility and cost. This is not merely theoretical; oral vaccines for polio and rotavirus have already shown the effectiveness of mucosal immunization at scale for enteric viral diseases. This review explores innate pattern recognition through TLR3, RIG-I, and MDA5 pathways, type I/III interferon (IFN) signaling, and mucosal SIgA responses across key EVs. It also assesses current mucosal vaccine candidates for EV-A71, CVA16, and EV-D68, most of which remain in preclinical stages. Understanding of these mechanistic gaps is important for advancing successful mucosal vaccine strategies into effective next-generation vaccines for EVs.
Intensive usage of cancer cell lines as cellular model system in cancer research caused the generation of variant subtypes during the last decades. Genetic aberrations, altered transcriptomic profiles as well as changed protein expression have been reported for diverse cancer cell lines resulting in hampered reproducibility of experimental results within the scientific community. Remarkably, the human leukemic THP-1 cell line, the most prominent model for immune cell research questions, was demonstrated to be a subject of branched evolution through diverse cultivation conditions; however, little is known about its impact on THP-1 as monocyte model. This study compares the whole transcriptomic landscape of THP-1 cells from two different depositors during differentiation into various immune states. Numerous genes showed significant altered transcript expression in immune-related signaling pathways, which pervade through macrophage differentiation. These differences are reflected in key biological processes such as metabolic pathways, including glycolysis and lipid metabolism. Furthermore, analysis of immune cell surface markers by flow cytometry and determination of cytokine levels using LEGENDplex revealed variations in protein expression, such as increased interleukin 10 (IL-10) raising questions about its impact on differentiation potential and immune function. These alterations have been evaluated by immune functional assays, including phagocytosis, determination of superoxide (SOX) levels as well as caspase-1 activity. In addition, the transcriptomic comparison to a primary immune cell dataset revealed high similarity between the THP-1 subtypes. However, minor discrepancies compared to primary cell types were identified, which could influence their immunological behavior. These findings suggest that the variability between THP-1 subtypes may undermine the reliability of THP-1-based immunological research, especially when attempting to replicate experiments across laboratories.
Dry eye disease (DED) can lead to severe eye discomfort, which manifests as dryness, pain and decreased vision, and significantly affects daily life. This study elucidated the effects of electroacupuncture (EA) intervention on DED and clarified its potential mechanisms through changes in lacrimal gland lipid metabolism and inflammation. A DED mouse model was established by repeated subcutaneous injections of scopolamine hydrobromide, followed by EA intervention. The therapeutic effects of EA intervention on DED were determined based on tear secretion volume (n = 10), corneal fluorescein staining scores (n = 10), corneal stromal (CS) damage (n = 6), and changes in corneal and lacrimal gland morphology (n = 10). Mechanistic endpoints included lacrimal gland lipidomic profiling by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) (n = 6) and assessment of IL-1β and IL-18 expression by immunohistochemistry (n = 6), western blotting (n = 3) and qRT-PCR (n = 6). EA increased tear secretion, reduced corneal fluorescein staining and ameliorated CS and lacrimal gland pathological alterations. Lipidomic analysis identified 81 metabolites, and Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analysis highlighted arachidonic acid metabolism and inflammation-related pathways. EA reduced arachidonic acid and several downstream pro-inflammatory metabolites while partially restoring omega-3 polyunsaturated fatty acids (PUFAs) and their derivatives. EA also reduced lacrimal gland IL-1β and IL-18 expression at the protein and mRNA levels. These findings suggest that EA may ameliorate scopolamine-induced DED, at least in part, through modulation of lacrimal gland lipid homeostasis and suppression of inflammatory responses.
Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide. Approximately 15% of localized and 5% of metastatic cases exhibit mismatch repair deficiency (dMMR) or high microsatellite instability (MSI-H). While immune checkpoint inhibitors (ICIs) have revolutionized the first-line treatment for this subgroup, 15%-46% of patients experience primary resistance, and a subset of responders eventually acquires resistance. This review synthesizes the multifaceted mechanisms underlying ICI resistance in dMMR/MSI-H CRC. We delineate tumor-intrinsic alterations, including defects in the antigen presentation machinery (specifically transporter associated with antigen processing [TAP]1/TAP2 and β2-microglobulin [β2m]), oncogenic signaling via the Wnt/β-catenin and JAK/STAT pathways, and epigenetic remodeling involving ARID1A. Furthermore, we explore the role of the immunosuppressive tumor microenvironment (TME), characterized by T-cell exclusion and myeloid-derived suppressor cell (MDSC) accumulation. To address these barriers, we evaluate the clinical potential of third-generation ICIs targeting lymphocyte activation gene 3 (LAG-3), T-cell immunoglobulin and mucin-domain-containing-3 (TIM-3), and TIGIT, as well as emerging biomarker strategies such as gut microbiome modulation and circulating tumor DNA (ctDNA) dynamics. By integrating these mechanistic insights with novel therapeutic approaches, including bispecific antibodies (BsAbs) and adoptive cell transfer, this review aims to provide a roadmap for overcoming resistance and advancing precision immunotherapy in dMMR/MSI-H CRC.
Objective Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by dynamically recurring inflammatory episodes, and targeting T cells has been shown to effectively alleviate RA. Caveolin‐1 (CAV‐1) is widely distributed in the membrane structures of immune cells and participates in the differentiation and homeostasis of CD4 + T cell subsets through signal transduction; however, its role in RA has not yet been investigated. This study aimed to examine the expression pattern and clinical relevance of CAV‐1 in RA, and to elucidate its regulatory role in CD4 + T cell‐mediated immune imbalance during disease progression. Methods This retrospective clinical study was conducted to detect serum CAV‐1 levels in RA and healthy controls (HCs), characterize its expression in peripheral blood CD4 + T cells and synovial tissues, and evaluate its association with clinical parameters. A collagen‐induced arthritis (CIA) mouse model was established and treated with the CAV‐1 inhibitor hydroxypropyl‐β‐cyclodextrin (HP‐β‐CD). Joint pathology, CD4 + T cell infiltration, differentiation, and the expression of related inflammatory cytokines were assessed using histopathological and immunological approaches. Results CAV‐1 expression in serum, CD4 + T cells, and synovial tissues was significantly higher in RA than in HC and was positively correlated with anti‐cyclic citrullinated peptide antibody (Anti‐CCP) and rheumatoid factor (RF) levels. HP‐β‐CD treatment markedly relieved joint inflammation and structural damage in CIA mice, reduced CD4 + T cell infiltration, and significantly suppressed the differentiation of T helper 1, T helper 17, and regulatory T (Treg) cells, accompanied by decreased expression of IFN‐γ, IL‐17A, and transforming growth factor‐β (TGF‐β). Conclusion CAV‐1 is aberrantly upregulated in RA and contributes to disease pathogenesis by regulating CD4 + T cell differentiation. Targeting CAV‐1 may represent a promising strategy for RA diagnosis and immunomodulatory therapy.
BACKGROUND:Chemical-induced sensitization is a major health concern. To date, no internationally accepted method is able to discriminate a skin from a respiratory sensitizer. The identification of an immune profile specific to skin or respiratory sensitizers is therefore scientifically relevant. METHODS:Female BALB/c mice were dermally exposed to a skin (2,4-dinitrochlorobenzene, DNCB) or to a respiratory sensitizer (phthalic anhydride, PA) at day 0 (D0) and D5 to initiate allergic sensitization and at D10, D11, and D12 to induce elicitation. Auricular lymph nodes (LNs) were collected at D0, D3, D7, D10, and D13. Single-cell transcriptomic and flow cytometric analyses were performed in order to identify immune cell proportions and marker expression. Enrichment analyses were performed at D13 on dendritic and T cells. Cytokine measurements were performed in LN cells' supernatant by bead array cytometry. RESULTS:Exposure to both sensitizers induced a strong immune reaction demonstrated by an increase of mature B cells and an evolution of the T cell subpopulations, with an increase of follicular and memory T cells. The proportion of NKT, Th2, and Th9 cells and the levels of IgE increased specifically during PA exposure. Follicular, memory, and helper T cells showed distinct transcriptomic responses to DNCB or PA. The proportion of dendritic cells (DCs) strongly increased starting from D10 in mice exposed to DNCB, and those cells showed a specific transcriptomic signature towards the skin sensitizer. Enrichment analyses suggested a metabolic shift in DCs exposed to DNCB. Single-cell transcriptomic data were confirmed by flow- and bead array cytometry. CONCLUSION:A distinct transcriptomic signature was identified in immune cells during sensitization to DNCB or PA in this study.
BACKGROUND:Allergic rhinitis (AR) is a prevalent chronic inflammatory nasal disorder with suboptimal current therapies. Glycyrrhizic acid (GA), a bioactive triterpenoid from Glycyrrhiza uralensis, exerts well-documented antiinflammatory effects, and preliminary evidence indicates its efficacy in alleviating airway inflammation in AR models. However, two critical knowledge gaps remain unaddressed: whether GA directly binds to core proteins of the toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB)/IL-1β inflammatory pathway, and whether this binding mediates GA's therapeutic effects in AR have not been systematically verified by integrated computational and in vivo experiments. METHODS:An ovalbumin (OVA)-induced AR rat model was established. GA was administered intranasally for seven consecutive days. Behavioral observations, histopathological examination, and serum cytokine detection were performed to evaluate AR-related symptoms and immune imbalance. Molecular docking was employed to assess the binding affinity between GA and four key proteins in the TLR4 pathway (TLR4, myeloid differentiation primary response 88 [MyD88], NF-κB, and IL-1β). About 100 ns molecular dynamics (MD) simulations were further conducted to validate the stability of the GA-protein complexes. Immunohistochemistry and RT-qPCR were used to verify the expression of pathway-related proteins and cytokines in nasal mucosal tissues. RESULTS:GA significantly reduced sneezing, rhinorrhea, and nasal mucosal pathological damage in AR rats. It restored the T helper 1 (Th1)/Th2 immune balance by suppressing Th2 cytokines (IL-4 and IL-13) and enhancing Th1 cytokines (interferon-gamma [IFN-γ] and IL-2). Molecular docking results showed that GA bound strongly to all four target proteins, with the highest affinity for IL-1β (-9.3 kcal/mol) and TLR4 (-7.6 kcal/mol). MD simulations confirmed the stable conformational dynamics of the GA-IL-1β and GA-TLR4 complexes. In vivo experiments further demonstrated that GA significantly downregulated the expression of TLR4, MyD88, NF-κB, and IL-1β in nasal mucosa and reduced the levels of downstream proinflammatory cytokines (TNF-α and IL-6). CONCLUSIONS:GA alleviates AR symptoms by directly binding to core proteins of the TLR4/NF-κB/IL-1β pathway, inhibiting pathway activation, restoring Th1/Th2 immune balance, and suppressing inflammatory responses. This study provides structural and functional evidence supporting GA as a promising targeted therapeutic candidate for AR.
OBJECTIVE:To clarify the regulatory mechanisms of the dynamic balance of histone H3 methylation in rheumatoid arthritis (RA) immune dysregulation and bone metabolism imbalance, and to explore related therapeutic approaches. METHODS:We systematically reviewed the roles of key histone H3 methylation sites, including H3K4me3, H3K9me3, and H3K27me3, in RA immune cell activation, inflammatory signaling pathway regulation, and bone cell differentiation. We analyzed the core functions of these sites in the cross-regulation of "immune dysregulation-bone metabolism imbalance" and discussed therapeutic strategies targeting this methylation balance. RESULTS:Accumulating preclinical evidence indicates that the dynamic balance of histone H3 methylation may regulate RA immune cell differentiation imbalance, excessive activation of inflammatory signaling pathways, and the imbalance state where bone resorption exceeds bone formation by modulating the activities of methyltransferases and demethylases. This balance appears to be a key mechanistic link connecting RA immune dysregulation and bone metabolism imbalance. Agents targeting this balance (such as EZH2 inhibitors and Janus kinase [JAK] inhibitors) have shown promising preclinical potential for dual anti-inflammatory and bone-protective effects. CONCLUSION:The dynamic balance of histone H3 methylation emerges as a potential core regulatory hub in the pathological process of RA based on preclinical studies, providing theoretical support for precise diagnosis and treatment of RA. Related targeting strategies may become a new direction to break through the limitations of existing treatments.
Neonatal acute respiratory distress syndrome (NARDS) is a life-threatening respiratory disorder characterized by high mortality and significant long-term morbidity. Increasing evidence suggests that pyroptosis contributes to pulmonary tissue injury in ARDS. In this study, single-cell RNA sequencing (scRNA-seq) was utilized to investigate the molecular mechanisms underlying pyroptosis within the NARDS microenvironment. Our analysis identified chemokine (C-C motif) ligand (CCL5) expression in M2 macrophages as a potential regulator of pyroptosis associated with impaired tissue repair. Mechanistically, CCL5 was linked to the activation of the pyroptotic pathway through interactions with inflammasome components, promoting caspase-11-dependent gasdermin D (GSDMD) cleavage and interleukin-1β (IL-1β) release. Network pharmacology analysis identified artesunate, a clinically approved antimalarial drug, as a potential inhibitor of CCL5 expression in patients with NARDS. Functional validation showed that artesunate attenuated M2 macrophage pyroptosis, reduced pulmonary injury, and promoted tissue repair. Collectively, our findings identify CCL5 as a potential regulator of macrophage pyroptosis and support the therapeutic potential of artesunate for modulating pyroptosis-associated inflammation in NARDS. This study provides a mechanistic framework for the development of CCL5-targeted interventions and highlights the dual anti-inflammatory and pro-repair properties of artesunate in NARDS treatment.
Endothelial cells (ECs) serve as crucial components of blood vessels and are vital for preserving vascular health in humans. The aging of these cells significantly accelerates vascular aging. Factors released by aging ECs serve as key initiators of arterial dysfunction and various cardiometabolic diseases. As a result, the targeted removal of aging ECs from injured tissues could reduce these issues and potentially improve lifespan. In this research, we explored the therapeutic capabilities of chimeric antigen receptor (CAR)-T cells designed to focus on senescent cells as potential senolytic agents. We identified GPNMB, a transmembrane glycoprotein, as a protein characterized by extensive expression during cellular senescence. Our findings demonstrate that CAR-T cells specifically targeting GPNMB can efficiently remove senescent cells in vitro. In addition to this, the engineered CAR-T cells significantly clear replicative and doxorubicin (Dox)-induced senescent human umbilical vein ECs (HUVECs). Notably, these CAR-T cells successfully eradicated senescent HUVECs within a Matrigel matrix implanted subcutaneously in mice, thereby creating a simulated in vivo environment that mimics physiological conditions. The findings emphasize the possibilities of CAR-T cells in treating EC senescence.
The identification of novel compounds capable of stimulating or suppressing immune responses is critical for advancing immunotherapy and improving our understanding of immune regulation. However, traditional screening approaches are often limited by low throughput, poor physiological relevance, and insufficient resolution to detect subtle immunomodulatory effects. In this study, we introduce a validated and robust multiplexed flow cytometry-based functional screening assay that enables efficient and precise identification of immunomodulatory compounds, while addressing the key limitations of conventional methods. By using human peripheral blood mononuclear cells (PBMCs) and defined stimulation conditions, the assay mimics complex immune interactions in a physiologically relevant context. It simultaneously monitors T- and B-cell responses across multiple functional readouts, including T-cell proliferation (carboxyfluorescein succinimidyl ester [CFSE]), activation (CD25, CD71), and B-cell costimulatory marker expression (CD86), enabling an in-depth assessment of immune modulation within heterogenous cell populations. The incorporation of robust statistical validation using the Z-factor further supports assay reliability and reproducibility. In a pilot screen with 295 immunology-related compounds, the assay demonstrated its ability to identify known immunomodulators with distinct functional profiles. This multiplex functional screening approach bridges the gap between mechanistic immunology and drug discovery by enabling systematic identification and characterization of immunotherapeutic candidates, while providing new insights into the complex dynamics of immune regulation.
