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Industrial multi-walled carbon nanotubes (ig-MWCNTs) make up the majority of carbon nanomaterials, and human contact with them is the most probable. At the same time, the biodegradation of ig-MWCNTs by phagocytes has not been studied—existing articles consider mainly laboratory-grade/functionalized MWCNTs (l-MWCNTs), in contrast to which ig-MWCNTs are a highly heterogeneous nanomaterial in terms of morphological and physicochemical characteristics. The aim of the present study was to analyze ig-MWCNTs' biodegradation by proinflammatory macrophages. We focused on both extra- and intracellular ig-MWCNTs' degradation. We analyzed biodegradation of two different types of ig-MWCNTs by human (THP-1) and murine (RAW264.7) macrophages. After 10 days of incubation, we studied nanoparticle localization within cells; isolated intra- and extracellular ig-MWCNTs were used for quantitative analysis. Ultrastructural and morphometric analysis were performed using transmission electron microscopy; electron diffraction was used for nanotube identification. To estimate chemical alterations, energy-dispersive X-ray spectroscopy and Raman spectroscopy were used. The study showed that both intra- and extracellular ig-MWCNTs undergo almost complete biodegradation, but in different ways: intracellular nanotubes become perforated and reduce to graphene flakes, while extracellular become thinner. We believe that the demonstrated variability in the destruction of ig-MWCNTs by cells suggests the possibility of creating nanomaterials with controlled biodegradation properties. [ABSTRACT FROM AUTHOR]

Industrial multi-walled carbon nanotubes (ig-MWCNTs) make up the majority of carbon nanomaterials, and human contact with them is the most probable. At the same time, the biodegradation of ig-MWCNTs by phagocytes has not been studied—existing articles consider mainly laboratory-grade/functionalized MWCNTs (l-MWCNTs), in contrast to which ig-MWCNTs are a highly heterogeneous nanomaterial in terms of morphological and physicochemical characteristics. The aim of the present study was to analyze ig-MWCNTs’ biodegradation by proinflammatory macrophages. We focused on both extra- and intracellular ig-MWCNTs’ degradation. We analyzed biodegradation of two different types of ig-MWCNTs by human (THP-1) and murine (RAW264.7) macrophages. After 10 days of incubation, we studied nanoparticle localization within cells; isolated intra- and extracellular ig-MWCNTs were used for quantitative analysis. Ultrastructural and morphometric analysis were performed using transmission electron microscopy; electron diffraction was used for nanotube identification. To estimate chemical alterations, energy-dispersive X-ray spectroscopy and Raman spectroscopy were used. The study showed that both intra- and extracellular ig-MWCNTs undergo almost complete biodegradation, but in different ways: intracellular nanotubes become perforated and reduce to graphene flakes, while extracellular become thinner. We believe that the demonstrated variability in the destruction of ig-MWCNTs by cells suggests the possibility of creating nanomaterials with controlled biodegradation properties.

Background: Nuclear factor κB (NF-κB) signaling in cardiac myocytes causes disease in a mouse model of arrhythmogenic cardiomyopathy (ACM) by mobilizing CCR2-expressing macrophages that promote myocardial injury and arrhythmias. Buccal mucosa cells exhibit pathologic features similar to those seen in cardiac myocytes in patients with ACM. Objectives: We sought to determine if persistent innate immune signaling via NF-κB occurs in cardiac myocytes in patients with ACM and if this is associated with myocardial infiltration of proinflammatory cells expressing CCR2. We also determined if buccal mucosa cells from young subjects with inherited disease alleles exhibit NF-κB signaling. Methods: We analyzed myocardium from ACM patients who died suddenly or required cardiac transplantation. We also analyzed buccal mucosa cells from young subjects with inherited disease alleles. The presence of immunoreactive signal for RelA/p65 in nuclei of cardiac myocytes and buccal cells was used as a reliable indicator of active NF-κB signaling. We also counted myocardial CCR2-expressing cells. Results: RelA/p65 signal was seen in numerous cardiac myocyte nuclei in 34 of 36 cases of ACM but not in 19 age-matched control individuals. Cells expressing CCR2 were increased in patient hearts in numbers directly correlated with the number of cardiac myocytes showing NF-κB signaling. NF-κB signaling was observed in buccal cells in young subjects with active disease. Conclusions: Patients with clinically active ACM exhibit persistent innate immune responses in cardiac myocytes and buccal mucosa cells, reflecting a local and systemic inflammatory process. Such individuals may benefit from anti-inflammatory therapy.

Clonal hematopoiesis (CH) is associated with elevated pro-inflammatory cytokines which act as a driving force for clonal expansion. Recurrent or chronic infections and sterile inflammation may predispose to CH, particularly in cases of germline predisposition. Blocking inflammation may be a viable therapeutic approach for high-risk CH and associated hematopoietic and non-hematopoietic health sequelae. Acquired genetic or cytogenetic alterations in a blood stem cell that confer clonal fitness promote its relative expansion leading to clonal hematopoiesis (CH). Despite a largely intact hematopoietic output, CH is associated with a heightened risk of progression to hematologic malignancies and with non-hematologic health manifestations, including cardiovascular disease and overall mortality. We focus on the evidence for the role of inflammation in establishing, maintaining and reciprocally being affected by CH. We describe the known pro-inflammatory signals associated with CH and preclinical studies that elucidated the cellular mechanisms involved. We review the evolving literature on early-onset CH in germline predisposition conditions and the possible role of immune dysregulation in this context. [ABSTRACT FROM AUTHOR]

Background. The effect on the activity of the multidrug resistance protein P-glycoprotein (P-gp, MDR1 gene) in pro-inflammatory (M1) human macrophages is considered one of the promising strategies for increasing the effectiveness of the treatment in patients with pulmonary tuberculosis: P-gp activity is considered a factor that reduces intracellular accumulation of rifampicin (RIF), a substrate for P-gp. The aim of this work was to reveal the effect of the therapeutic concentration of RIF on the activity of P-gp in M1 human macrophages. The objectives were as follows: to determine the expression levels of the MDR1 gene, P-gp protein, as well as its functional activity at different periods of cell differentiation and under the influence of RIF.Material and methods. The following cell lines were used in the work: suspension cells of promonocytic leukemia THP-1 and THP-1 macrophages induced by phorbol ether according to the pro-inflammatory phenotype. Suspension cells of myeloid leukemia K562/IS-9 transfected with the MDR1 gene were used as a comparison group. An important factor is the choice of the experimental concentration of RIF: the average concentration of the drug in patients with pulmonary tuberculosis was 10 µg/ml. The methods of RT-PCR, immunocytochemistry, and flow cytometry were used in the work.Results and discussion. The induction of MDR1 gene expression in M1 macrophages under short-term exposure to a therapeutic concentration of RIF was revealed. This effect is typical only for THP-1 macrophages, in which a significant functional activity of P-gp is registered. This induction does not occur in the cells with no detectable P-gp activity (THP-1 suspension cells). This indicates the presence of different mechanisms of RIF influence on MDR1, which can be used to develop a strategy for P-gp inhibition in inflammatory macrophages.Conclusion. Given the key role of macrophages in tuberculosis, further evaluation of MDR1/P-gp in the surgical material of patients with pulmonary tuberculosis is necessary, which makes it possible to draw a conclusion that it is necessary to develop and apply drug strategies aimed at blocking the functional activity of P-gp and choosing more effective anti-tuberculosis therapy regimens.

There is a paucity of human models to study immune-mediated host damage. Here, we utilized the GeoMx spatial multi-omics platform to analyze immune cell changes in COVID-19 pancreatic autopsy samples, revealing an accumulation of proinflammatory macrophages. Single-cell RNA sequencing (scRNA-seq) analysis of human islets exposed to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) or coxsackievirus B4 (CVB4) viruses identified activation of proinflammatory macrophages and β cell pyroptosis. To distinguish viral versus proinflammatory-macrophage-mediated β cell pyroptosis, we developed human pluripotent stem cell (hPSC)-derived vascularized macrophage-islet (VMI) organoids. VMI organoids exhibited enhanced marker expression and function in both β cells and endothelial cells compared with separately cultured cells. Notably, proinflammatory macrophages within VMI organoids induced β cell pyroptosis. Mechanistic investigations highlighted TNFSF12-TNFRSF12A involvement in proinflammatory-macrophage-mediated β cell pyroptosis. This study established hPSC-derived VMI organoids as a valuable tool for studying immune-cell-mediated host damage and uncovered the mechanism of β cell damage during viral exposure., Competing Interests: Declaration of interests R.E.S. is on the scientific advisory board of Miromatrix Inc. and Lime Therapeutics and is a consultant and speaker for Alnylam Inc. S.C. and T.E are the co-founders of OncoBeat, LLC. S.C. is a consultant of Vesalius Therapeutics and co-founder of iOrganBio., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

The problem of reduced effectiveness of antituberculosis drugs is currently associated not only with the development of drug resistance in the pathogen but also with the functional activity of the P-gp (P-glycoprotein) multidrug resistance protein of macroorganism cells. One of the main antituberculosis drugs, rifampicin (RIF), is a substrate for P-gp, which reduces its effectiveness. The article presents the data on the P-gp activity in proinflammatory human macrophages and assesses the effect of a new form of RIF encapsulated in lactic acid polymer nanoparticles, approved for medical use, on its activity. It was shown that THP-1 macrophages, in contrast to the THP-1 monocytes, are characterized by the functional activity increase of P-gp during differentiation as well as the preservation of the viability in the presence of RIF, which is associated with the P-gp activation. The new form of RIF is nontoxic to THP-1 monocytes and macrophages in comparison with the traditional form. When using the encapsulated form of RIF, the process of endocytosis/phagocytosis in macrophages is activated and the functional activity of P-gp decreases. The data obtained demonstrate that the development of encapsulated antituberculosis drugs, phagocytosis activators, which targetedly reduce the P-gp activity and thus affect the functional characteristics of human macrophages, seems to be a promising and relevant direction for increasing the effectiveness of antituberculosis drugs. [ABSTRACT FROM AUTHOR]

(1→3)-β-D-glucans (BG) (the glucose polymers) are recognized as pathogen motifs, and different forms of BGs are reported to have various effects. Here, different BGs, including Pachyman (BG with very few (1→6)-linkages), whole-glucan particles (BG with many (1→6)-glycosidic bonds), and Oat-BG (BG with (1→4)-linkages), were tested. In comparison with dextran sulfate solution (DSS) alone in mice, DSS with each of these BGs did not alter the weight loss, stool consistency, colon injury (histology and cytokines), endotoxemia, serum BG, and fecal microbiome but Pachyman–DSS-treated mice demonstrated the highest serum cytokine elicitation (TNF-α and IL-6). Likewise, a tail vein injection of Pachyman together with intraperitoneal lipopolysaccharide (LPS) induced the highest levels of these cytokines at 3 h post-injection than LPS alone or LPS with other BGs. With bone marrow-derived macrophages, BG induced only TNF-α (most prominent with Pachyman), while LPS with BG additively increased several cytokines (TNF-α, IL-6, and IL-10); inflammatory genes (iNOS, IL-1β, Syk, and NF-κB); and cell energy alterations (extracellular flux analysis). In conclusion, Pachyman induced the highest LPS proinflammatory synergistic effect on macrophages, followed by WGP, possibly through Syk-associated interactions between the Dectin-1 and TLR-4 signal transduction pathways. Selection of the proper form of BGs for specific clinical conditions might be beneficial. [ABSTRACT FROM AUTHOR]

Studies of the role of macrophages in phagocytosis are of great theoretical and practical importance for understanding how these cells are involved in the organism’s defense response and in the development of various pathologies. Here we investigated phagocytic plasticity of THP-1 (acute monocytic human leukemia) cells at different stages (days 1, 3, and 7) of phorbol ester (PMA)-induced macrophage differentiation. Analysis of cytokine profiles showed that PMA at a concentration of 100 nM induced development of the proinflammatory macrophage population. The functional activity of macrophages was assessed on days 3 and 7 of differentiation using unlabeled latex beads and latex beads conjugated with ligands (gelatin, mannan, and IgG Fc fragment) that bind to the corresponding specific receptors. The general phagocytic activity increased significantly (1.5-2.0-fold) in the course of differentiation; phagocytosis occurred mostly through the Fc receptors, as shown previously for M1 macrophages. On day 7, the levels of phagocytosis of gelatin-and Fc-covered beads were high; however, the intensity of ingestion of mannan-conjugated beads via mannose receptors increased 2.5-3.0-fold as well, which indicated formation of cells with an alternative phenotype similar to that of M2 macrophages. Thus, the type and the plasticity of phagocytic activity at certain stages of macrophage differentiation can be associated with the formation of functionally mature morphological phenotype. This allows macrophages to exhibit their phagocytic potential in response to specific ligands. These data are of fundamental importance and can be used to develop therapeutic methods for correcting the M1/M2 macrophage ratio in an organism. [ABSTRACT FROM AUTHOR]

Inflammatory bowel disease (IBD) is a chronic disease with an unclear pathogenesis for which successful treatments are still lacking. It has been reported that procyanidin, a natural antioxidant, relieves colitis, but the specific mechanism is elusive. Our present study was designed to investigate the effects of procyanidin on colitis and the regulation of the M1 macrophage phenotype and related signaling pathways. In vivo, we used two classic colitis models to observe the effect of procyanidin on macrophage polarization. In vitro, we further validated the therapeutic effect of procyanidin in the RAW264.7 cell line and peritoneal macrophages. The current findings provide new evidence that procyanidin ameliorated dextran sulfate sodium (DSS)-induced colitis by preventing the polarization of macrophages to the M1 type and downregulating the levels of proinflammatory factors in cells. We also showed that procyanidin prevented lipopolysaccharide (LPS)-induced elevation of inflammatory cytokines and the activation of proinflammatory macrophages, which was achieved by activating the STAT3 and NF-κB pathways. This is the first study to demonstrate that procyanidin alleviates experimental colitis by inhibiting the polarization of proinflammatory macrophages. These data reveal new ideas for the pathogenesis and treatment of inflammatory diseases. [Display omitted] • Procyanidin may act as a prophylactic against DSS-induced colitis. • Procyanidin improves colitis by inhibiting the activation of M1 macrophages. • Procyanidin inhibites inflammation by downregulating the NF-κB and STAT3 pathways. [ABSTRACT FROM AUTHOR]

Background: Nuclear factor κB (NF-κB) signaling in cardiac myocytes causes disease in a mouse model of arrhythmogenic cardiomyopathy (ACM) by mobilizing CCR2-expressing macrophages that promote myocardial injury and arrhythmias. Buccal mucosa cells exhibit pathologic features similar to those seen in cardiac myocytes in patients with ACM., Objectives: We sought to determine if persistent innate immune signaling via NF-κB occurs in cardiac myocytes in patients with ACM and if this is associated with myocardial infiltration of proinflammatory cells expressing CCR2. We also determined if buccal mucosa cells from young subjects with inherited disease alleles exhibit NF-κB signaling., Methods: We analyzed myocardium from ACM patients who died suddenly or required cardiac transplantation. We also analyzed buccal mucosa cells from young subjects with inherited disease alleles. The presence of immunoreactive signal for RelA/p65 in nuclei of cardiac myocytes and buccal cells was used as a reliable indicator of active NF-κB signaling. We also counted myocardial CCR2-expressing cells., Results: RelA/p65 signal was seen in numerous cardiac myocyte nuclei in 34 of 36 cases of ACM but not in 19 age-matched control individuals. Cells expressing CCR2 were increased in patient hearts in numbers directly correlated with the number of cardiac myocytes showing NF-κB signaling. NF-κB signaling was observed in buccal cells in young subjects with active disease., Conclusions: Patients with clinically active ACM exhibit persistent innate immune responses in cardiac myocytes and buccal mucosa cells, reflecting a local and systemic inflammatory process. Such individuals may benefit from anti-inflammatory therapy., (© 2023 Heart Rhythm Society. Published by Elsevier Inc.)

Background: Heterogeneous macrophages play an important role in multiple liver diseases, including viral fulminant hepatitis (VFH). Fibrinogen-like protein 2 (FGL2) is expressed on macrophages and regulates VFH pathogenesis; however, the underlying mechanism remains unclear., Aim: To explore how FGL2 regulates macrophage function and subsequent liver injury during VFH., Methods: Murine hepatitis virus strain 3 (MHV-3) was used to induce VFH in FGL2-deficient ( Fgl2-/- ) and wild-type (WT) mice. The dynamic constitution of hepatic macrophages was examined. Adoptive transfer of Fgl2-/- or WT bone marrow-derived macrophages (BMDMs) into WT recipients with macrophages depleted prior to infection was carried out and the consequent degree of liver damage was compared. The signaling cascades that may be regulated by FGL2 were detected in macrophages., Results: Following MHV-3 infection, hepatic macrophages were largely replenished by proinflammatory monocyte-derived macrophages (MoMFs), which expressed high levels of FGL2. In Fgl2-/- mice, the number of infiltrating inflammatory MoMFs was reduced compared with that in WT mice after viral infection. Macrophage depletion ameliorated liver damage in WT mice and further alleviated liver damage in Fgl2-/- mice. Adoptive transfer of Fgl2-/- BMDMs into macrophage-removed recipients significantly reduced the degree of liver damage. Inhibition of monocyte infiltration also significantly ameliorated liver damage. Functionally, Fgl2 deletion impaired macrophage phagocytosis and the antigen presentation potential and attenuated the proinflammatory phenotype. At the molecular level, FGL2 deficiency impaired IRF3, IRF7, and p38 phosphorylation, along with NF-κB activation in BMDMs in response to viral infection., Conclusion: Infiltrated MoMFs represent a major source of hepatic inflammation during VFH progression, and FGL2 expression on MoMFs maintains the proinflammatory phenotype via p38-dependent positive feedback, contributing to VFH pathogenesis., Competing Interests: Conflict-of-interest statement: All authors declare no conflict of interest in the study., (©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.)

Background : Exosome (Exo)-based chemotherapeutic drug delivery systems have been extensively investigated; however, the therapeutic potential of other subtypes of extracellular vesicles (EVs), in particular microvesicles (MiV), seem to be overlooked. Moreover, despite a general agreement on organ tropism of EVs, few studies have clearly demonstrated that EVs specifically target tumor tissue. Methods : Proinflammatory macrophage-derived EV subpopulations comprising apoptotic bodies (ApB), MiV and Exo were isolated under differential ultracentrifugation, and further analyzed using comparative proteomic and lipid approach. Results : On the basis of EV biogenesis pathways, our data demonstrated that MiV acquire the tumor-targeting capacity probably through inheritance of CCR2-enriched cell membrane which also drives the recruitment of donor cells to tumor sites. Further, our data validate MiV utilize SNARE-mediated membrane fusion to directly discharge doxorubicin to nucleus and bypass endocytic degradation. Conclusions : Compared with other EV subtypes, MiV loaded with doxorubicin gain significant benefits in chemotherapeutic outcomes including survival rate improvements in metastatic ovarian cancer. Therefore, MiV represent a potent alterative to Exo and synthetic liposomes (Lipo) for tumor-targeting drug delivery., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)