Inhibition of Triggering Receptor Expressed on Myeloid Cell-1 Alleviates Acute Gouty Inflammation

Gout is a prevalent form of aseptic inflammation caused by the deposition of monosodium urate (MSU) crystals in joints or tissues. Triggering receptor expressed on myeloid cell-1 (TREM-1) is a superimmunoglobulin receptor expressed on innate immune cells including granulocytes, monocytes, and macrophages. TREM-1 serves as a link between innate immunity and adaptive immunity, playing a crucial role in regulating inflammation and immune response. The purpose of this study was to investigate the potential role of TREM-1 in THP-1 cells and peripheral blood mononuclear cells (PBMCs) from patients with gouty arthritis (GA). In the current study, we found that the mRNA and protein levels of TREM-1 increased in PBMCs from GA patients and soluble TREM-1 in plasma as well. In addition, an increased level of TREM-1 was observed in THP-1 treated with monosodium urate (MSU) in vitro, along with upregulation of proinflammatory cytokines. Moreover, upon specific inhibition of TREM-1, Toll-like receptor 4 (TLR-4), and myeloid differentiation factor 88 (MyD88), the levels of MyD88 and proinflammatory cytokines were decreased after MSU challenge in THP-1 cells. Interestingly, inhibition of TLR-4 could enhance the effect of TREM-1 inhibitor in MSU-induced inflammation. Taken together, our findings suggested that TREM-1 could accelerate MSU-induced acute inflammation. Inhibition of TREM-1 may provide a new strategy for alleviating acute gouty inflammation.

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H3K23/H3K36 hypoacetylation and HDAC1 up-regulation are associated with adverse consequences in obstructive sleep apnea patients

Scientific Reports ◽

10.1038/s41598-021-00052-9 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Yung-Che Chen ◽

Po-Yuan Hsu ◽

Chien-Hung Chin ◽

Chang-Chun Hsiao ◽

Chia-Wei Liou ◽

...

Keyword(s):

Obstructive Sleep Apnea ◽

Sleep Apnea ◽

Mononuclear Cells ◽

Gene Promoter ◽

Peripheral Blood Mononuclear ◽

Protein Levels ◽

Reactive Protein ◽

Primary Snoring ◽

Obstructive Sleep

AbstractThe aim of this study is to determine the roles of global histone acetylation (Ac)/methylation (me), their modifying enzymes, and gene-specific histone enrichment in obstructive sleep apnea (OSA). Global histone modifications, and their modifying enzyme expressions were assessed in peripheral blood mononuclear cells from 56 patients with OSA and 16 matched subjects with primary snoring (PS). HIF-1α gene promoter-specific H3K36Ac enrichment was assessed in another cohort (28 OSA, 8 PS). Both global histone H3K23Ac and H3K36Ac expressions were decreased in OSA patients versus PS subjects. H3K23Ac expressions were further decreased in OSA patients with prevalent hypertension. HDAC1 expressions were higher in OSA patients, especially in those with excessive daytime sleepiness, and reduced after more than 6 months of continuous positive airway pressure treatment. H3K79me3 expression was increased in those with high C-reactive protein levels. Decreased KDM6B protein expressions were noted in those with a high hypoxic load, and associated with a higher risk for incident cardiovascular events or hypertension. HIF-1α gene promoter-specific H3K36Ac enrichment was decreased in OSA patients versus PS subjects. In vitro intermittent hypoxia with re-oxygenation stimuli resulted in HDAC1 over-expression and HIF-1α gene promoter-specific H3K36Ac under-expression, while HDAC1 inhibitor, SAHA, reversed oxidative stress through inhibiting NOX1. In conclusions, H3K23/H3K36 hypoacetylation is associated with the development of hypertension and disease severity in sleep-disordered breathing patients, probably through up-regulation of HDAC1, while H3K79 hypermethylation is associated with higher risk of cardiovascular diseases, probably through down-regulation of KDM6B.

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Decreased miR-4512 Levels in Monocytes and Macrophages of Individuals With Systemic Lupus Erythematosus Contribute to Innate Immune Activation and Neutrsophil NETosis by Targeting TLR4 and CXCL2

Frontiers in Immunology ◽

10.3389/fimmu.2021.756825 ◽

2021 ◽

Vol 12 ◽

Author(s):

Binbin Yang ◽

Xinwei Huang ◽

Shuangyan Xu ◽

Li Li ◽

Wei Wu ◽

...

Keyword(s):

Systemic Lupus Erythematosus ◽

Lupus Erythematosus ◽

Target Genes ◽

Mononuclear Cells ◽

Therapeutic Potential ◽

Luciferase Reporter ◽

Systemic Lupus ◽

Peripheral Blood Mononuclear ◽

Monocytes And Macrophages

ObjectiveSystemic lupus erythematosus (SLE) is an autoimmune disease with complex etiology that is not yet entirely understood. We aimed to elucidate the mechanisms and therapeutic potential of microRNAs (miRNAs) in SLE in a Tibetan population.MethodsPeripheral blood mononuclear cells from SLE patients (n = 5) and healthy controls (n = 5) were used for miRNA–mRNA co-sequencing to detect miRNAs related to immune abnormalities associated with SLE. Luciferase reporter assay was used to identify potential targets of candidate miRNA. The target genes were verified in miRNA-agomir/antagomir transfection assays with multiple cells lines and by expression analysis. The effects of candidate miRNA on monocyte and macrophage activation were evaluated by multiple cytokine profiling. Neutrophil extracellular traps (NETs) formation was analyzed in vitro by cell stimulation with supernatants of monocytes and macrophages transfected with candidate miRNA. The rodent MRL/lpr lupus model was used to evaluate the therapeutic effect of CXCL2Ab on SLE and the regulation effect of immune disorders.ResultsIntegrated miRNA and mRNA expression profiling identified miRNA-4512 as a candidate miRNA involved in the regulation of neutrophil activation and chemokine-related pathways. MiR-4512 expression was significantly reduced in monocytes and macrophages from SLE patients. MiR-4512 suppressed the TLR4 pathway by targeting TLR4 and CXCL2. Decreased monocyte and macrophage miR-4512 levels led to the expression of multiple proinflammatory cytokines in vitro. Supernatants of miR-4512 antagomir-transfected monocytes and macrophages significantly promoted NETs formation (P < 0.05). Blocking of CXCL2 alleviated various pathogenic manifestations in MRL/lpr mice, including kidney damage and expression of immunological markers of SLE.ConclusionsWe here demonstrated the role of miR-4512 in innate immunity regulation in SLE. The effect of miR-4512 involves the regulation of monocytes, macrophages, and NETs formation by direct targeting of TLR4 and CXCL2, indicating the miR-4512-TLR4-CXCL2 axis as a potential novel therapeutic target in SLE.

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Crystals of monosodium urate monohydrate enhance lipopolysaccharide-induced release of interleukin 1β by mononuclear cells through a caspase 1-mediated process

Annals of the Rheumatic Diseases ◽

10.1136/ard.2007.082222 ◽

2008 ◽

Vol 68 (2) ◽

pp. 273-278 ◽

Cited By ~ 77

Author(s):

E J Giamarellos-Bourboulis ◽

M Mouktaroudi ◽

E Bodar ◽

J van der Ven ◽

B-J Kullberg ◽

...

Keyword(s):

Mononuclear Cells ◽

Gouty Arthritis ◽

Underlying Mechanism ◽

Monosodium Urate ◽

Peripheral Blood Mononuclear ◽

Caspase 1 ◽

Pyrin Domain ◽

Factor Α ◽

Msu Crystals ◽

Urate Crystals

Objective:Recent studies suggest that crystals of monosodium urate (MSU), deposited in joints of patients with acute gouty arthritis, activate the NACHT domain, leucine-rich repeat and pyrin domain-containing protein (NALP)3 inflammasome. In the present study we have investigated whether production of proinflammatory cytokines by crystals was exacerbated during costimulation with Toll-like receptor (TLR) ligands.Methods:Mononuclear cells of 22 healthy donors were stimulated by various concentrations of MSU crystals in the absence or presence of lipopolysaccharide (LPS), Pam3Cys and flagellin. Production of tumour necrosis factor α (TNFα), interleukin (IL)1β and IL6, as well as the intracellular concentrations of proIL1β were measured by ELISA. mRNA transcripts of TNFα and IL1β were assessed by real-time PCR. Stimulation experiments were also performed with peripheral blood mononuclear cells (PBMCs) of one patient carrying a NALP3 mutation.Results:MSU induced a moderate release of IL1β and IL6, but not of TNFα. Urate crystals amplified IL1β production stimulated by the TLR4 ligand LPS, while no synergy was apparent for IL6 production. In addition, no synergy between urate crystals and Pam3Cys (TLR2 ligand) or flagellin (TLR5 ligand) was apparent. The synergy between urate crystals and LPS was directed at the level of the NALP3 inflammasome, as it was present only when active IL1β was measured, but not at the level of IL1 mRNA or proIL1β. The synergy between LPS and MSU crystals ceased to exist in the presence of a caspase 1 inhibitor.Conclusions:MSU crystals act in synergy with LPS for the induction of enhanced release of IL1β. Increased cleavage of proIL1β by urate-activated caspase 1 is proposed as the underlying mechanism.

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Constitutive Activation of STATs Upon In Vivo Human Immunodeficiency Virus Infection

Blood ◽

10.1182/blood.v94.12.4202 ◽

1999 ◽

Vol 94 (12) ◽

pp. 4202-4209 ◽

Cited By ~ 54

Author(s):

Chiara Bovolenta ◽

Laura Camorali ◽

Alessandro L. Lorini ◽

Silvia Ghezzi ◽

Elisa Vicenzi ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Mononuclear Cells ◽

Janus Kinase ◽

Peripheral Blood Mononuclear ◽

Constitutive Activation ◽

Protein Levels ◽

Immunodeficiency Virus ◽

Stat Pathway

Abstract Infection by the human immunodeficiency virus (HIV) either upregulates or downregulates the expression of several cytokines and interferons (IFNs) that use the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway for signal transduction. However, very little is known on the state of activation of the JAK/STAT pathway after HIV infection either in vivo or in vitro. In this regard, we report here that a constitutive activation of a C-terminal truncated STAT5 (STAT5▵) and of STAT1 occurs in the majority (∼75%) of individuals with progressive HIV disease. We have further demonstrated that, among peripheral blood mononuclear cells (PBMCs), STAT5▵ is activated preferentially in CD4+ T cells. In contrast to a published report, expression of STATs from PBMCs of infected individuals was comparable with that of seronegative donors. In addition, in vitro infection of mitogen-activated PBMCs with a panel of laboratory-adapted and primary HIV strains characterized by differential usage of chemokine coreceptors did not affect STAT protein levels. However, enhanced activation of STAT was observed after in vitro infection of resting PBMCs and nonadherent PBMCs by different viral strains. Thus, constitutive STAT activation in CD4+T lymphocytes represents a novel finding of interest also as a potential new marker of immunological reconstitution of HIV-infected individuals.

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Cytokine production during in vitro hemodialysis with new and formaldehyde- or renalin-reprocessed cellulose dialyzers.

Journal of the American Society of Nephrology ◽

10.1681/asn.v641304 ◽

1995 ◽

Vol 6 (4) ◽

pp. 1304-1308

Author(s):

B J Pereira ◽

B Snodgrass ◽

G Barber ◽

C Perella ◽

S Chopra ◽

...

Keyword(s):

Proinflammatory Cytokines ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Cytokine Production ◽

Peripheral Blood Mononuclear ◽

Blood Compartment ◽

Alpha Production ◽

Blood Mononuclear Cells

Critics of reuse have suggested that patients treated with reprocessed dialyzers are exposed to pyrogen trapped from the water or solutions used during the reprocessing cycle, thereby triggering the synthesis and release of proinflammatory cytokines, resulting in cachexia. To test this hypothesis, the production of interleukin (IL)-1 alpha by peripheral blood mononuclear cells (PBMC) during in vitro dialysis with new or reprocessed cellulose dialyzers was compared. An in vitro closed-loop dialysis circuit was created with standard hemodialysis blood lines and either new cellulose dialyzers or dialyzers reprocessed 10 times with either formaldehyde/bleach (formaldehyde) or peracetic acid/hydrogen peroxide mixture (Renalin). The circuit was rinsed with 2 L or more of pyrogen-free normal saline before the start of in vitro dialysis until the blood compartment tested negative for residual formaldehyde/Renalin. Heparinized whole blood from healthy volunteers was circulated for 3 h in the blood compartment at 100 mL/min at 37 degrees C. The dialysate compartment was sealed. Peripheral blood mononuclear cells (PBMC) were harvested from the blood compartment before and at the end of 3 h of in vitro dialysis. Total IL-1 alpha synthesis (cell associated plus secreted) by unstimulated and endotoxin-stimulated PBMC was measured by a specific, non-cross-reactive radioimmunoassay. After 3 h of in vitro dialysis, IL-1 alpha production (in picograms per 2.5 million PBMC) by unstimulated PBMC increased from 354 +/- 63 at baseline to 454 +/- 57 with new dialyzers (P = 0.25), from 453 +/- 101 to 450 +/- 67 with formaldehyde-reprocessed dialyzers (P = 0.98), and from 360 +/- 61 to 538 +/- 144 with Renalin-reprocessed dialyzers (P = 0.23). IL-1 alpha production by endotoxin-stimulated PBMC increased from 5,214 +/- 996 to 9,237 +/- 929 with new dialyzers (P < = 0.001), from 6,395 +/- 955 to 9,636 +/- 1,058 with formaldehyde-reprocessed dialyzers (P = 0.006), and from 7,561 +/- 1,000 to 10,092 +/- 2,470 with Renalin-reprocessed dialyzers (P = 0.32). However, there were no significant differences among groups with respect to IL-1 alpha production by unstimulated or endotoxin-stimulated PBMC either before or after 3 h of in vitro dialysis. These data argue against the suggestion that exposure to reprocessed dialyzers results in enhanced synthesis of proinflammatory cytokines. In fact, reprocessed dialyzers probably induce less cytokine production than do new cellulose dialyzers.

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Fingolimod reduces circulating tight-junction protein levels and in vitro peripheral blood mononuclear cells migration in multiple sclerosis patients

Scientific Reports ◽

10.1038/s41598-018-33672-9 ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 4

Author(s):

Pasquale Annunziata ◽

Chiara Cioni ◽

Gianni Masi ◽

Maristella Tassi ◽

Giuseppe Marotta ◽

...

Keyword(s):

Multiple Sclerosis ◽

Tight Junction ◽

Peripheral Blood Mononuclear Cells ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Protein Levels ◽

Junction Protein ◽

Blood Mononuclear Cells ◽

Multiple Sclerosis Patients

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The Staphyloccous aureus Eap Protein Activates Expression of Proinflammatory Cytokines

Infection and Immunity ◽

10.1128/iai.01699-07 ◽

2008 ◽

Vol 76 (5) ◽

pp. 2164-2168 ◽

Cited By ~ 15

Author(s):

Thomas J. Scriba ◽

Sophie Sierro ◽

Eric L. Brown ◽

Rodney E. Phillips ◽

Andrew K. Sewell ◽

...

Keyword(s):

Proinflammatory Cytokines ◽

Mononuclear Cells ◽

Ex Vivo ◽

Human Peripheral Blood ◽

Intercellular Adhesion Molecule ◽

Intercellular Adhesion Molecule 1 ◽

Peripheral Blood Mononuclear ◽

Necrosis Factor Alpha ◽

Tnf Α ◽

Monocytes And Macrophages

ABSTRACT The extracellular adhesion protein (Eap) secreted by the major human pathogen Staphylococcus aureus is known to have several effects on human immunity. We have recently added to knowledge of these roles by demonstrating that Eap enhances interactions between major histocompatibility complex molecules and human leukocytes. Several studies have indicated that Eap can induce cytokine production by human peripheral blood mononuclear cells (PBMCs). To date, there has been no rigorous attempt to identify the breadth of cytokines produced by Eap stimulation or to identify the cell subsets that respond. Here, we demonstrate that Eap induces the secretion of the proinflammatory cytokines interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) by CD14+ leukocytes (monocytes and macrophages) within direct ex vivo PBMC populations (note that granulocytes are also CD14+ but are largely depleted from PBMC preparations). Anti-intercellular adhesion molecule 1 (CD54) antibodies inhibited this induction and implicated a role for this known Eap binding protein in cellular activation. IL-6 and TNF-α secretion by murine cells exposed to Eap was also observed. The activation of CD14+ cells by Eap suggests that it could play a significant role in both septic shock and fever, two of the major pathological features of S. aureus infections.

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Comparative Investigation of Frankincense Nutraceuticals: Correlation of Boswellic and Lupeolic Acid Contents with Cytokine Release Inhibition and Toxicity against Triple-Negative Breast Cancer Cells

Nutrients ◽

10.3390/nu11102341 ◽

2019 ◽

Vol 11 (10) ◽

pp. 2341 ◽

Cited By ~ 5

Author(s):

Schmiech ◽

Lang ◽

Ulrich ◽

Werner ◽

Rashan ◽

...

Keyword(s):

Breast Cancer ◽

Triple Negative Breast Cancer ◽

Proinflammatory Cytokines ◽

Triple Negative ◽

Mononuclear Cells ◽

Breast Cancer Cell Lines ◽

Peripheral Blood Mononuclear ◽

Tnf Α

For centuries, frankincense extracts have been commonly used in traditional medicine, and more recently, in complementary medicine. Therefore, frankincense constituents such as boswellic and lupeolic acids are of considerable therapeutic interest. Sixteen frankincense nutraceuticals were characterized by high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS), revealing major differences in boswellic and lupeolic acid compositions and total contents, which varied from 0.4% to 35.7%. Frankincense nutraceuticals significantly inhibited the release of proinflammatory cytokines, such as TNF-α, IL-6, and IL-8, by LPS-stimulated peripheral blood mononuclear cells (PBMC) and whole blood. Moreover, boswellic and lupeolic acid contents correlated with TNF-α, IL-1β, IL-6, IL-8, and IL-10 inhibition. The nutraceuticals also exhibited toxicity against the human triple-negative breast cancer cell lines MDA-MB-231, MDA-MB-453, and CAL-51 in vitro. Nutraceuticals with total contents of boswellic and lupeolic acids >30% were the most active ones against MDA-MB-231 with a half maximal inhibitory concentration (IC50) ≤ 7.0 µg/mL. Moreover, a frankincense nutraceutical inhibited tumor growth and induced apoptosis in vivo in breast cancer xenografts grown on the chick chorioallantoic membrane (CAM). Among eight different boswellic and lupeolic acids tested, β-ABA exhibited the highest cytotoxicity against MDA-MB-231 with an IC50 = 5.9 µM, inhibited growth of cancer xenografts in vivo, and released proinflammatory cytokines. Its content in nutraceuticals correlated strongly with TNF-, IL-6, and IL-8 release inhibition.

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Constitutive Activation of STATs Upon In Vivo Human Immunodeficiency Virus Infection

Blood ◽

10.1182/blood.v94.12.4202.424k22_4202_4209 ◽

1999 ◽

Vol 94 (12) ◽

pp. 4202-4209 ◽

Cited By ~ 4

Author(s):

Chiara Bovolenta ◽

Laura Camorali ◽

Alessandro L. Lorini ◽

Silvia Ghezzi ◽

Elisa Vicenzi ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Mononuclear Cells ◽

Janus Kinase ◽

Peripheral Blood Mononuclear ◽

Constitutive Activation ◽

Protein Levels ◽

Immunodeficiency Virus ◽

Stat Pathway

Infection by the human immunodeficiency virus (HIV) either upregulates or downregulates the expression of several cytokines and interferons (IFNs) that use the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway for signal transduction. However, very little is known on the state of activation of the JAK/STAT pathway after HIV infection either in vivo or in vitro. In this regard, we report here that a constitutive activation of a C-terminal truncated STAT5 (STAT5▵) and of STAT1 occurs in the majority (∼75%) of individuals with progressive HIV disease. We have further demonstrated that, among peripheral blood mononuclear cells (PBMCs), STAT5▵ is activated preferentially in CD4+ T cells. In contrast to a published report, expression of STATs from PBMCs of infected individuals was comparable with that of seronegative donors. In addition, in vitro infection of mitogen-activated PBMCs with a panel of laboratory-adapted and primary HIV strains characterized by differential usage of chemokine coreceptors did not affect STAT protein levels. However, enhanced activation of STAT was observed after in vitro infection of resting PBMCs and nonadherent PBMCs by different viral strains. Thus, constitutive STAT activation in CD4+T lymphocytes represents a novel finding of interest also as a potential new marker of immunological reconstitution of HIV-infected individuals.

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