Search Results (628)

(TB) induces a pronounced thromboinflammatory state during active disease, characterized by elevated fibrinogen, D-dimer, and thrombin-related activity, reduced levels of endogenous anticoagulants, impaired fibrinolysis, platelet activation, and endothelial dysfunction. Although many of these abnormalities improve after treatment initiation, accumulating evidence suggests that microbiological cure may not fully restore vascular, immune, and hemostatic homeostasis. This raises the possibility that TB leaves a persistent thromboinflammatory imprint after cure. This narrative synthesizes current evidence on tuberculosis-associated hypercoagulability during active disease and after treatment, and proposes a conceptual framework for post-tuberculosis thromboinflammatory memory grounded in cellular persistence, tissue remodeling, and extracellular vesicle-mediated signaling. Candidate storage compartments include hematopoietic stem and progenitor cells, monocyte/macrophage lineages, alveolar macrophages, remodeled pulmonary endothelium, and fibrotic post-TB lung tissue. EVs may function as mobile vectors that transfer procoagulant phospholipids, tissue factor, inflammatory proteins, and regulatory microRNAs between these compartments, thereby linking local post-TB remodeling to systemic vascular and coagulation pathways. A mechanistic evidence ladder is proposed, encompassing phenotypic persistence, EV cell-of-origin attribution, molecular persistence, paired longitudinal validation, functional transfer, and clinical outcome linkage. Current data support the biological plausibility of this framework but remain insufficient to establish post-TB thromboinflammatory memory as a defined clinical entity. Direct evidence in long-term TB survivors is still lacking, particularly with respect to persistent EV signatures, cell-specific reservoirs, and the functional transfer of procoagulant phenotypes. Longitudinal, cell-resolved, multi-omic, and functionally validated studies are required to determine whether TB leaves a durable thromboinflammatory memory, where it is stored, and whether it contributes to long-term thrombotic and cardiovascular risk. This article should be interpreted as a narrative review with a conceptual framework rather than as evidence that post-tuberculosis thromboinflammatory memory is already a formally established clinical entity. Full article

Dysregulated activation of the NLRP3 inflammasome is a key driver in the pathogenesis of numerous inflammatory disorders. This study aimed to evaluate the protective effect of Platycodon grandiflorus polysaccharide (PGPS_t) against NLRP3-inflammasome-mediated inflammation and elucidate its underlying mechanisms. An in vitro inflammatory model was established in porcine alveolar macrophages (3D4/21) using LPS/ATP co-stimulation. The effects of PGPS_t were assessed by measuring inflammasome activation, intracellular reactive oxygen species (ROS) generation, and pro-inflammatory cytokine secretion. Molecular docking, alongside inhibitors (NAC, MCC950) and siRNA targeting NEK7, was employed to probe the involved mechanisms. PGPS_t significantly suppressed NLRP3 inflammasome assembly and activation, reduced caspase-1 cleavage, and decreased the maturation and release of IL-1β and IL-18. It exerted its inhibitory effects through dual mechanisms: scavenging intracellular ROS and directly binding to NEK7 and NLRP3 to disrupt their interaction, as supported by molecular docking. The anti-inflammatory effect was diminished upon NEK7 knockdown. In conclusion, PGPS_t is an effective natural inhibitor of the NLRP3 inflammasome, functioning through ROS clearance and direct interference with the NLRP3–NEK7 interaction. These findings propose PGPS_t as a promising therapeutic candidate and further validate NEK7 as a potential target for treating NLRP3-driven inflammatory diseases. Full article

Periodontitis is a chronic inflammatory disease characterized by alveolar bone loss driven by dysregulated immune responses. We previously showed that extracellular vesicles derived from retinoic acid-induced regulatory T lymphocytes (RA-Treg EVs) suppress pathogenic CD4⁺ T-lymphocyte responses and reduce alveolar bone loss during periodontitis. Herein, we investigated whether RA-Treg EVs also modulate macrophage and CD8⁺ T-lymphocyte responses during experimental periodontitis. Ligature-induced periodontitis was generated in mice, followed by local administration of RA-Treg EVs. Alveolar bone loss was analyzed by micro-computed tomography, and periodontal tissues and cervical lymph nodes were analyzed by flow cytometry to quantify antigen-presenting cells, macrophages, macrophage subsets, and CD8⁺ T lymphocytes. The direct effects of RA-Treg EVs on macrophage phenotype and CD8⁺ T-cell proliferation and activation were assessed in vitro. RA-Treg EV treatment attenuated alveolar bone loss and preserved trabecular microarchitecture. This effect was associated with reduced macrophage infiltration into periodontal tissues, modulation of macrophage polarization, and restoration of CD8⁺ T-cell abundance in periodontal tissues and draining cervical lymph nodes, without major changes in CD8⁺IFN-γ⁺ or CD8⁺RANKL⁺ cells. In vitro, RA-Treg EVs induced heterogeneous macrophage phenotypes distinct from the classical M1/M2 polarization states while markedly enhancing CD8⁺ T-cell proliferation and activation. These findings indicate that RA-Treg EVs preserve alveolar bone during experimental periodontitis while selectively modulating macrophage and CD8⁺ T-lymphocyte responses. Full article

Hemorrhagic pneumonia is a severe and often fatal disease in captive forest musk deer (Moschus berezovskii), but the pathogen remains incompletely understood. Based on incomplete statistics, the estimated incidence in captive populations ranges from 20% to 80%, with the disease occurring mainly in autumn, winter, and early spring. The disease has an acute onset and rapid progression. Due to the species’ strong stress response, affected animals rarely show behavioral changes, making early detection difficult. In this study, we investigated a mortality case presenting with oral bleeding and hematemesis on a forest musk deer farm. Postmortem examination revealed diffuse hemorrhagic pneumonia, and lung tissue samples were collected for histopathology, bacterial isolation, full-length 16S rRNA gene sequencing, and DNA/RNA virome sequencing. Histological examination showed extensive alveolar hemorrhage, fibrinous exudate, and macrophage infiltration. Bacterial culture and 16S rRNA gene sequencing identified Bergeyella zoohelcum as the predominant bacterium, accounting for 100% of the bacterial community in the lung tissue. Virome analysis revealed predominantly DNA bacteriophages (e.g., Cirlivirales, Cremevirales, Microviridae) and no known pathogenic RNA viruses; only seven low-abundance, unclassified RNA viral contigs of low completeness were detected. These results indicate that B. zoohelcum is the likely causative agent of hemorrhagic pneumonia in this case, with no evidence of viral involvement. This study provides the first direct association of B. zoohelcum with hemorrhagic pneumonia in forest musk deer, highlighting its pathogenic potential and the importance of monitoring this bacterium in captive populations. Full article

Old murine bone marrow-derived monocytes and macrophages (BMMs) display enhanced CD38 protein, a nicotinamide adenine dinucleotide (NAD⁺) glycohydrolase, and reduced NAD⁺ level after infection with oral pathogens compared to young controls. We aimed to determine whether treatment with a CD38-specific inhibitor (78c) in mice with experimental periodontitis could alleviate alveolar bone loss and enhance NAD⁺ levels in tissues compared with vehicle treatment. Twenty young (2-month-old) and twenty old (18-month-old) male C57BL/6J mice with experimental periodontitis were treated with either vehicle or 78c twice daily via intraperitoneal injection for 4 weeks. The liver, spleen, and right maxillary tissues were harvested to analyze NAD⁺ levels. The left maxillary tissues were scanned by micro-CT. Then, the left maxillary tissues were processed for tissue sectioning and stained with hematoxylin and eosin (H&E) and tartrate−resistant acid phosphatase (TRAP). Treatment with 78c significantly enhanced NAD⁺ levels in the liver and spleen of both young and old mice, and significantly increased NAD⁺ in the right maxilla of old mice compared with vehicle treatment. Additionally, treatment with 78c alleviated alveolar bone loss in both young and old mice. Our results support the notion that 78c is a promising therapeutic strategy for treating periodontal disease. Full article

Porcine reproductive and respiratory syndrome (PRRS) is an immunosuppressive disease caused by PRRS virus (PRRSV). PRRSV infection not only compromises the host immune defenses, but also predisposes the host to secondary infections by other pathogens, of which PRV is one of the common secondary infection pathogens. Porcine alveolar macrophages (PAMs) are the primary target cells of PRRSV, and their phagocytic function is critical for immune defense, homeostasis maintenance, and disease regulation. However, PRRSV disrupts PAMs phagocytosis, impairing the host’s ability to combat infection. This study used PRV-pAb complexes as phagocytic indicators, investigated the effect of PRRSV infection on PAMs phagocytosis and its underlying molecular mechanisms. We found that PRRSV infection interfered with phagosome maturation—a process regulated by Rab7 and other regulators, thereby blocking phagocytic degradation and significantly suppressing PAMs phagocytic activity. Further analysis revealed that reactive oxygen species (ROS) play a key role in this process. Elevated ROS levels damaged lysosomal membrane integrity, ultimately inhibiting phagosome-lysosome fusion. Notably, phagocytosis of PRRSV-infected PAMs was partially restored with N-acetylcysteine (NAC) by reducing ROS levels. These findings offer novel insights into PRRSV-induced immunosuppression and secondary infections while providing a theoretical foundation for developing more effective PRRSV prevention and control strategies. Full article

Local immune cell activation and vascular remodelling are characteristic pathogenic features of pulmonary arterial hypertension (PAH). HIV and schistosome infections have been individually associated with PAH. However, whether co-infection with these pathogens has a distinct impact on the development of pulmonary vascular disease remains poorly understood, partly due to the lack of experimental animal models. In a novel non-infectious model of HIV and Schistosoma pulmonary co-exposure based on lung embolisation of S. mansoni eggs in HIV-transgenic (HIV) mice, we previously reported exacerbated endothelial remodelling and dysfunction, along with increased pulmonary arterial pressure; which were associated with a unique profile of pro-inflammatory cytokines in the lung. In the present study, we used flow cytometric analysis of isolated lung leukocytes and immunofluorescence staining to characterise the pulmonary immune cell landscape associated with individual or combined exposure to HIV and schistosome. Compared with mice exposed to HIV (untreated HIV mice) or schistosome (egg-treated wild-type mice), co-exposed (egg-treated HIV mice) animals showed significantly increased numbers of interstitial and alveolar macrophages, patrolling-type monocytes, NKT and γδ T cells, and reduced CD8⁺ αβ T cells. Other lung immune cells, including inflammatory-type monocytes, eosinophils/neutrophils, dendritic cells, CD4⁺ αβ T cells, NK cells and B cells were not significantly affected in the co-exposure condition. Taken together, these results show for the first time that combined pulmonary exposure to HIV and Schistosoma, as it may occur in co-infected individuals, alters the local immune cell landscape in a manner distinct from that of individual exposure. Furthermore, these findings may contribute to a better understanding of the complex inflammatory processes involved in the pathogenesis of PAH, thereby supporting the development of therapies targeting pathogenic immune cells in pulmonary vascular disease associated with HIV and Schistosoma co-morbidity. Full article

Deoxynivalenol (DON) is a mycotoxin commonly found in food crops and animal feed worldwide. Its pronounced toxicity in pigs poses a serious risk to the swine industry and to human health. This study focused on two central features of ferroptosis—iron metabolism and lipid peroxidation—and examined how chlorogenic acid (CGA) affects DON-induced ferroptosis in porcine alveolar macrophages (PAMs) via cell-based assays and oxidative lipid metabolomics. These findings show that DON disrupts intracellular iron homeostasis by altering iron-handling proteins (upregulating TFR1 and DMT1 and downregulating FPN1), which may lead to iron overload. Concurrently, DON impairs the GPX4 antioxidant axis (downregulating GPX4, SLC3A2, SLC7A11, and GCLC) and increases ROS, and exposure led to a significant increase in numerous oxidized lipid metabolites, consistent with elevated lipid peroxidation, culminating in ferroptosis in PAMs. CGA mitigates these effects by restoring iron homeostasis and reestablishing GPX4 axis function, thereby reducing oxidative stress. Moreover, CGA suppresses lipid peroxidation pathways, notably linoleic acid oxidation metabolism. In conclusion, CGA protects PAMs and mitigates the proferroptotic effects of DON. Full article

Objective: To establish a porcine complement receptor type 1-like (CR1-like)-mediated targeted anti-porcine reproductive and respiratory syndrome virus (PRRSV) nanodrug delivery system by investigating interactions between erythrocytes from Landrace piglets (both male and female), PRRSV, and anti-PRRSV nanodrugs. Methods: Optimal conditions for PRRSV sensitization with fresh porcine serum were determined. CR1-like-dependent immune adhesion of porcine erythrocytes to sensitized PRRSV was verified by immunofluorescence, electron microscopy, qPCR, and Western blot. The effect of this adhesion on PRRSV infection of porcine alveolar macrophages (PAMs) was studied using a flow chamber system. Mannose-modified matrine nanoliposomes (MMLNPs) were prepared, characterized, and evaluated for cytotoxicity, targeting ability, and in vitro antiviral activity. Results: PRRSV was optimally sensitized by incubation with fresh porcine serum at 37 °C for 2 h. Porcine erythrocytes specifically adhered to sensitized PRRSV via CR1-like, significantly promoting PRRSV infection of PAMs. Stable, uniform-sized MMLNPs showed no cytotoxicity, targeted PAMs via CR1-like, and exhibited superior antiviral activity to free matrine. Conclusions: CR1-like-mediated immune adhesion is a critical mechanism for PRRSV infection of PAMs. Harnessing this natural pathway enables efficient targeted delivery of matrine nanoliposomes to PAMs, providing a promising translational strategy for PRRSV control. Full article

Periodontitis-induced destruction of periodontal tissues and tooth loss remain major clinical challenges. Although periodontal regenerative therapies aim to reconstruct damaged structures, particularly the repair of alveolar bone defects, current biomaterials have limited capacity to simultaneously promote cell adhesion and osteogenic differentiation. Amelogenin (Am) plays a key role in mineralized tissue development through its highly conserved N-terminal and C-terminal regions, but studies have shown that Am has certain limitations in promoting cell adhesion. In contrast, the arginine-glycine-aspartic acid (RGD) domain of fibronectin (FN) effectively mediates cell–matrix adhesion. Based on these properties, we developed a novel recombinant fusion protein (rtAmR) by combining the conserved regions of Am with the RGD domain of FN. In vitro, rtAmR significantly promoted the adhesion and spreading of human stem cells from the apical papilla (hSCAPs) compared with the control group. Quantitative analysis showed that the number of adherent cells and the cell spreading area in the rtAmR group were 1.9-fold and 2.1-fold higher than those in the rhAm group, respectively. In osteogenic differentiation assays, rtAmR exhibited activity comparable to that of rhAm and even outperformed rhAm in terms of alkaline phosphatase (ALP) activity, collagen type I (COL I) expression, and calcium nodule formation. In a Sprague–Dawley (SD) rat alveolar bone defect model, rtAmR treatment significantly promoted bone regeneration, achieving superior bone volume/total volume (BV/TV) values compared to the rhAm and rhFN groups. Immunohistochemistry revealed that rtAmR did not obviously increase neutrophils, mast cells, or M2 macrophages versus control, confirming its biosafety and suggesting M2-independent osteogenesis. These findings suggest that rtAmR is a promising bifunctional bioactive protein for periodontal bone regeneration. Full article

SIRT2 (Sirtuin 2) is an NAD+-dependent deacetylase that exerts crucial regulatory effects on immune homeostasis and macrophage activation. While chronic cold exposure is a known predisposing factor for pulmonary dysfunction, the precise mechanisms by which SIRT2 potentially modulates lung macrophage polarization under cold stress remains poorly understood. In this study, we evaluated the protective capacity of SIRT2 using both wild-type (WT) and Sirt2-knockout (Sirt2−/−) murine models subjected to chronic cold exposure (4 °C for 3 h daily over 21 days). Our results demonstrated that Sirt2 deficiency significantly exacerbated cold-induced pulmonary histopathological damage and increased the secretion of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) (p < 0.05). Furthermore, chronic cold stress triggered a macrophage-centered inflammatory response, a process wherein SIRT2 was found to curtail M1 pro-inflammatory polarization. To further investigate these mechanisms, in vitro experiments were conducted using the mouse alveolar macrophage cell line MH-S. While LPS was utilized as a canonical inflammatory stimulus to mimic the injury environment, SIRT2 overexpression was found to reverse the LPS-induced increase in M1 markers and attenuate inflammatory cytokine secretion. These findings suggest that SIRT2 maintains intracellular homeostasis by modulating macrophage plasticity and plays a protective role in the development of chronic cold stimulus-induced lung injury. Consequently, SIRT2 activation may represent a potential therapeutic pathway for the treatment of environment-related respiratory diseases. Full article

Efferocytosis—the tightly regulated clearance of apoptotic cells by phagocytes—maintains tissue homeostasis and is impaired in chronic obstructive pulmonary disease (COPD), where it contributes to persistent inflammation and increases the risk of comorbidities, including lung cancer. Inhaled corticosteroids (ICS) and long-acting β2 agonists (LABAs) are cornerstones of COPD therapy, but their effects on efferocytosis and on the COPD–lung cancer interface are incompletely understood. The primary objective of this study was to determine whether the ICS fluticasone propionate and the LABA salmeterol xinafoate, alone or in combination at clinically informed concentrations (10⁻⁸–10⁻⁶ M; 10⁻⁴ M reserved for cytotoxicity screening), modulate efferocytic capacity and inflammatory cell survival across diverse phagocyte models. We performed standardized in vitro efferocytosis assays using murine peritoneal and alveolar macrophages, the murine macrophage line J774A.1, PMA-differentiated human THP-1 macrophages, human blood-derived neutrophils, and the human alveolar adenocarcinoma cell line A549. Apoptosis was induced in Jurkat T cells by UV irradiation (100 mJ/cm²) and in murine thymocytes by dexamethasone (1 µM, 4 h); apoptotic and necrotic populations were characterized by annexin-V/propidium iodide and Sytox Green/Hoechst H-33342 staining. Peritoneal macrophages showed the highest efferocytic activity (~75%), followed by J774A.1 (~75% at 24 h), THP-1 (~30% at 2 h; ~60% at 24 h), alveolar macrophages (~40%), and A549 cells (<20%). Neither fluticasone nor salmeterol, individually or in combination, significantly altered efferocytic capacity in any phagocyte tested (all ANOVA p > 0.26). Fluticasone (10⁻⁸ and 10⁻⁶ M) significantly improved 24 h neutrophil survival and reduced early apoptosis (p < 0.05) but did not translate this survival benefit into enhanced efferocytosis. Salmeterol was cytotoxic at 10⁻⁴ M and inactive at 10⁻⁸–10⁻⁶ M. These findings indicate that the established anti-inflammatory benefits of ICS/LABA in COPD do not extend to augmentation of efferocytosis in this acute, serum-free in vitro setting and that pharmacological restoration of efferocytosis in COPD—a defect implicated in the pathogenesis and progression of comorbid lung cancer—will likely require strategies targeting the efferocytic machinery itself (e.g., MerTK, Rac-1, MFG-E8) rather than relying on current inhaled therapy. Full article

African swine fever virus (ASFV) is the causative agent of African swine fever (ASF), a fatal and highly contagious disease, resulting in enormous losses to the global swine industry. No licensed vaccines or effective therapeutics are currently available to control ASFV infection. Interferons (IFNs) serve as key mediators of host antiviral immunity by inducing interferon-stimulated genes (ISGs), but the specific mechanisms by which individual ISGs restrict ASFV replication remain unclear. Interferon-induced protein with tetratricopeptide repeats 3 (IFIT3, also called ISG60) has been shown to exhibit antiviral activity against various viruses, but its role in ASFV infection has not been previously studied. Here, we used porcine alveolar macrophages (PAMs), the primary target cells of ASFV, to investigate IFIT3’s function in ASFV replication. We found that overexpression of IFIT3 inhibited ASFV replication, while its knockdown enhanced viral propagation. Mechanistically, IFIT3 directly blocked ASFV adsorption to host cells, thereby suppressing all subsequent stages of the viral cycle. IFIT3 also specifically interacted with ASFV F334L, an early viral gene product that encodes the small subunit of ribonucleotide reductase, a key enzyme for viral DNA synthesis. Additionally, IFIT3 positively regulated the STAT1/TBK1/IRF3 signaling axis: its overexpression increased phosphorylation of TBK1 and IRF3, as well as the protein level of STAT1, while IFIT3 knockdown attenuated activation of these molecules. Transcriptomic analysis of IFIT3-knockout PAMs revealed significant suppression of innate immune pathways, including type I interferon, JAK-STAT, and RIG-I-like receptor pathways, along with downregulated expression of core antiviral molecules such as ISG15, MX1, and STAT1. Conversely, pathways related to viral adsorption, endocytosis, and cytoskeleton were activated, and pathways involved in protein translation initiation, endoplasmic reticulum stress, and autophagy were dysregulated, creating a favorable intracellular environment for ASFV replication. In conclusion, IFIT3 restricts ASFV replication possibly by inhibiting viral adsorption and promoting innate immune signaling, identifying it as a potential therapeutic target against ASFV. This study’s limitation is its in vitro PAM model; future work will validate IFIT3’s role in vivo and develop targeted inhibitors. Full article

Concurrent alcohol and cannabis use (“crossfading”) is increasingly prevalent, especially among adolescents, yet its toxicological impact on pulmonary innate immunity remains largely unexplored. Alveolar macrophages (AMs) orchestrate inflammatory responses in the lung, and dysregulated macrophage polarization is a hallmark of alcohol-associated lung disease. Although alcohol and cannabinoids individually modulate immune function, the mechanisms by which their co-exposure alters macrophage activation and inflammatory signaling in the lung are largely unknown. AMs are highly sensitive to xenobiotic exposure and play a central role in regulating inflammatory and cytotoxic responses. In this study, we investigated how acute ethanol exposure, synthetic cannabinoid exposure, and their combined exposure affect macrophage viability, polarization, and the release of inflammatory mediators via cannabinoid receptor (CB1R/CB2R)-dependent pathways. Human THP-1-derived macrophages and KG-1 macrophage-like cells were exposed to ethanol, the CB1/CB2 agonist WIN 55,212-2, or both, with selective pharmacological antagonism of CB1R and CB2R. Ethanol exposure activated and polarized macrophages toward a pro-inflammatory M1 phenotype, accompanied by increased secretion of pro-inflammatory cytokines MCP-1, TGF-α, IFN-β, IL-6, and TNF-α. In contrast, WIN 55,212-2 promoted anti-inflammatory M2 polarization and increased IL-10 and IL-4 production. Notably, co-exposure to ethanol and WIN produced an antagonistic immunomodulatory response, characterized by the suppression of ethanol-induced M1 polarization and attenuation of pro-inflammatory cytokine release. Mechanistically, pharmacological CB1R blockade reduced ethanol-induced M1 polarization and cytokine secretion, whereas CB2R blockade exacerbated these effects, underscoring divergent roles for cannabinoid receptors in regulating pulmonary macrophage responses. This study provides novel findings demonstrating the mechanism by which alcohol–cannabinoid co-use reshapes macrophage immune phenotypes and identifies the endocannabinoid system as a potential therapeutic target for alcohol-related inflammatory lung disease. Full article

The secondary immunomodulatory effects of conventional therapeutics, such as antibiotics and cytostatics, are frequently overlooked despite their significant clinical implications. Building on our previous findings that drugs like paclitaxel and doxorubicin heavily influence macrophage polarization—potentially driving metastasis or inflammation—this study systematically evaluates the secondary immune-modulating actions of standard drugs and natural adjuvants. Using patient-derived bronchoalveolar lavage (BAL) fluid (ex vivo alveolar macrophages), we developed an analytical platform using synthetic carbohydrate-functionalized fluorescent ligands targeting key receptors (CD206, CD209, CD280, CD301). Integrating ligand-binding profiles with Linear Discriminant Analysis (LDA) yielded quantitative immune-state vectors capable of differentiating favorable and unfavorable prognostic signatures and imbalanced immune states. Pro-filing samples across heterogeneous respiratory conditions revealed highly con-text-dependent responses. While some treatments synergistically corrected unfavorable imbalanced profiles, others provoked dysregulation. Notably, in pneumonia or bronchitis with an asthma-prone M2-dominant profile, specific antibiotic regimens are critical; doxycycline, for instance, may exacerbate patient deterioration by further driving M2a polarization. Crucially, we identified that natural adjuvants (e.g., curcumin, coumarins, polyphenols) exhibit potent properties capable of correcting these adverse secondary drug effects. Ultimately, this profiling platform highlights the necessity of evaluating patient-specific secondary drug effects, offering a functional blueprint for precision immunotherapy and the rational design of adjuvant-enhanced treatments. Full article