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BACKGROUND: Oleoylethanolamide (OEA), an endogenously generated cannabinoid-like compound, has been reported to be increased in patients with severe asthma and aspirin-exacerbated respiratory disease. Recruitment of activated eosinophils in the airways is a hallmark of bronchial asthma. OBJECTIVE: We explored the direct contribution of cannabinoid receptor 2 (CB2), a cognate receptor of OEA, which induces eosinophil activation in vitro and in vivo. METHODS: We investigated OEA signaling in the eosinophilic cell line dEol-1 in peripheral blood eosinophils from people with asthma. In order to confirm whether eosinophil activation by OEA is CB2 dependent or not, CB2 small interfering RNA and the CB2 antagonist SR144528 were used. The numbers of airway inflammatory cells and the levels of cytokines were measured in bronchoalveolar lavage fluid, and airway hyperresponsiveness was examined in the BALB/c mice. RESULTS: CB2 expression was increased after OEA treatment in both peripheral blood eosinophils and dEol-1 cells. It was also elevated after OEA-induced recruitment of eosinophils to the lungs in vivo. However, SR144528 treatment reduced the activation of peripheral blood eosinophils from asthmatic patients. Furthermore, CB2 knockdown decreased the activation of dEol-1 cells and the levels of inflammatory and type 2 cytokines. SR144528 treatment alleviated airway hyperresponsiveness and eosinophil recruitment to the lungs in vivo. CONCLUSION: CB2 may contribute to the pathogenesis of eosinophilic asthma. Our results provide new insight into the molecular mechanism of signal transduction by OEA in eosinophilic asthma.
Assuntos
Asma , Canfanos , Endocanabinoides , Ácidos Oleicos , Eosinofilia Pulmonar , Pirazóis , Receptor CB2 de Canabinoide , Animais , Humanos , Camundongos , Asma/metabolismo , Citocinas , Inflamação/patologia , Pulmão/patologia , Ácidos Oleicos/metabolismo , Eosinofilia Pulmonar/metabolismo , Receptores de Canabinoides , Receptor CB2 de Canabinoide/metabolismoRESUMO
BACKGROUND: Neutrophilic asthma (NA) is a severe asthma phenotype associated with steroid resistance and IL-1ß overproduction; however, the exact mechanism remains unclear. Moreover, the dysfunction of TNF-α signaling pathway, a regulator of IL-1ß production, was associated with the deficiency of ovarian tumor protease deubiquitinase with linear linkage specificity (otulin) in autoimmune patients. OBJECTIVE: We hypothesized that otulin downregulation in macrophages (Mφ) could trigger Mφ activation via the nucleotide-binding domain, leucine-rich repeat, and pyrin domain-containing protein 3 (NLRP3) inflammasome signaling pathway. METHODS: We assessed the expressions of otulin in blood monocyte subsets from NA patients and in alveolar Mφ from NA mice. Additionally, we evaluated the functional consequences of otulin deficiency in bone marrow-derived Mφ. The effects of inhibiting receptor-interacting protein kinase (RIPK)-1 and RIPK-3 on neutrophils and group 3 innate lymphoid cells (ILC3s) were assessed in vitro and in vivo. RESULTS: When comparing nonclassical monocytes, a significant downregulation of otulin in the intracellular components was observed in NA patients compared to healthy controls (P = .005). Moreover, isolated alveolar Mφ from the NA mice exhibited lower otulin expression compared to those from control mice. After otulin knockdown in bone marrow-derived Mφ, we observed spontaneous IL-1ß production depending on NLRP3 inflammasome. Moreover, the infiltrated neutrophils and ILC3s were significantly decreased by combined treatment of RIPK-1 and RIPK-3 inhibitors through blocking IL-1ß release in NA. CONCLUSIONS: IL-1ß overproduction caused by a deficiency of otulin, an upstream triggering factor, could be a promising diagnostic and therapeutic target for NA.
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BACKGROUND: Lactobacillus paracasei has been known to reduce airway resistance and inflammation in asthma. However, the therapeutic effect of its extracellular vesicles (EVs) in patients with asthma remains unclear. METHODS: To validate the clinical relevance of L. paracasei-derived EVs (LpEV) in asthma, the composition of gut microbial EVs was verified by metagenomics in LPS-induced C57BL/6 mice. The components of proteins and metabolites in LpEV were identified by peptide mass fingerprinting and metabolomic analysis. The serum levels of specific IgG1 or IgG4 antibodies to LpEV were compared by ELISA between patients with eosinophilic asthma (EA, n = 10) and those with neutrophilic asthma (NA, n = 10) as well as with healthy controls (HCs, n = 10). Finally, therapeutic effects of LpEV and their metabolites in asthma were validated in vivo/in vitro. RESULTS: Significantly lower proportions of EVs derived from Lactobacillus at the genus level were noted in mice with NA than in control mice. Moreover, the serum levels of LpEV-specific IgG4, but not IgG1, were lower in patients with NA than in those with EA or in HCs and positively correlated with FEV1 (%) values. In addition, oral administration of LpEV reduced airway resistance and inflammation in mice with NA. Finally, LpEV and their 3 metabolites (dodecanoic acid, palmitoleic acid, and D-(-)-tagatose) significantly inhibited JNK phosphorylation/IL-8 production in airway epithelium in vitro. CONCLUSIONS: These findings suggest that LpEV may have a therapeutic potential targeting NA by suppressing the JNK pathway and proinflammatory cytokine production in airway epithelium.
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Asma , Vesículas Extracelulares , Lacticaseibacillus paracasei , Animais , Humanos , Camundongos , Asma/terapia , Modelos Animais de Doenças , Epitélio , Imunoglobulina G , Inflamação , Pulmão , Sistema de Sinalização das MAP Quinases , Camundongos Endogâmicos C57BLRESUMO
BACKGROUND: Increased blood/sputum neutrophil counts are related to poor clinical outcomes of severe asthma (SA), where we hypothesized that classical monocytes (CMs)/CM-derived macrophages (Mφ) are involved. We aimed to elucidate the mechanisms of how CMs/Mφ induce the activation of neutrophils/innate lymphoid cells (ILCs) in SA. METHODS: Serum levels of monocyte chemoattractant protein-1 (MCP-1) and soluble suppression of tumorigenicity 2 (sST2) were measured from 39 patients with SA and 98 those with nonsevere asthma (NSA). CMs/Mφ were isolated from patients with SA (n = 19) and those with NSA (n = 18) and treated with LPS/interferon-gamma. Monocyte/M1Mφ extracellular traps (MoETs/M1ETs) were evaluated by western blotting, immunofluorescence, and PicoGreen assay. The effects of MoETs/M1ETs on neutrophils, airway epithelial cells (AECs), ILC1, and ILC3 were assessed in vitro and in vivo. RESULTS: The SA group had significantly higher CM counts with increased migration as well as higher levels of serum MCP-1/sST2 than the NSA group. Moreover, the SA group had significantly greater production of MoETs/M1ETs (from CMs/M1Mφ) than the NSA group. The levels of MoETs/M1ETs were positively correlated with blood neutrophils and serum levels of MCP-1/sST2, but negatively correlated with FEV1%. In vitro/in vivo studies demonstrated that MoETs/M1ETs could activate AECs, neutrophils, ILC1, and ILC3 by increased migration as well as proinflammatory cytokine production. CONCLUSIONS: CM/Mφ-derived MoETs/M1ETs could contribute to asthma severity by enhancing neutrophilic airway inflammation in SA, where modulating CMs/Mφ may be a potential therapeutic option.
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Asma , Armadilhas Extracelulares , Humanos , Monócitos , Imunidade Inata , Linfócitos , Neutrófilos , Inflamação , MacrófagosRESUMO
BACKGROUND: Platelets play a modulatory role in inflammatory response by secreting a vast array of granules and disintegrating into membrane-bound microparticles upon activation. The interplay between eosinophils and platelets is postulated to be implicated in the pathology of allergic airway inflammation. In this study, we investigated whether activated platelets can induce eosinophil extracellular trap (EET) formation, a cellular process by which activated eosinophils release net-like DNA fibers. METHODS: Platelets were stimulated with the calcium ionophore, A23187, and the platelet agonists, thrombin and adenosine diphosphate (ADP). Platelet cultures were fractionated into conditioned medium (CM) and pellet, which were then overlaid on eosinophils to examine EET formation. RESULTS: The CM and pellet from A23187-activated platelets stimulated eosinophils to generate EET, whereas those from thrombin- or ADP-activated platelets failed to induce such generation. The EET-inducing activity of the A23187-activated platelet culture was linearly proportional to the number of activated platelets. Interestingly, while EET formation induced by the direct stimulation of eosinophils with A23187 was NADPH oxidase (NOX)-dependent, EET formation induced by A23187-activated platelets was NOX-independent and significantly inhibited by necroptosis pathway inhibitors. CONCLUSIONS: Activated platelets and their products may induce EET formation, thereby potentiating their role in eosinophilic airway inflammation.
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Plaquetas , Armadilhas Extracelulares , Humanos , Plaquetas/metabolismo , Trombina/farmacologia , Trombina/metabolismo , Ionóforos de Cálcio/metabolismo , Calcimicina/farmacologia , Calcimicina/metabolismo , Armadilhas Extracelulares/metabolismo , Inflamação/metabolismo , Difosfato de Adenosina/metabolismo , Cálcio/metabolismoRESUMO
BACKGROUND: Genetic variants of dipeptidyl peptidase 10 (DPP10) have been suggested to contribute to the development of NSAID-exacerbated respiratory disease (NERD). However, the mechanisms of how DPP10 contributes to NERD phenotypes remain unclear. OBJECTIVE: To demonstrate the exact role of DPP10 in the pathogenesis of NERD. METHODS: Patients with NERD (n = 110), those with aspirin-tolerant asthma (ATA, n = 130) and healthy control subjects (HCs, n = 80) were enrolled. Clinical characteristics were analysed according to the serum DPP10 levels in both NERD and ATA groups. The function of DPP10 in airway inflammation and remodelling was investigated with in vitro, ex vivo and in vivo experiments. RESULTS: NERD patients had higher levels of serum DPP10 and TGF-ß1 with lower FEV1 than ATA patients or HCs (p < .05 for each). NERD patients with higher DPP10 levels had higher TGF-ß1, but lower FEV1 (p < .05 for all), whilst no differences were noted in ATA patients. Moreover, the seum DPP10 levels had a positive correlation with TGF-ß1 (r = 0.384, p < .001), but a negative correlation with FEV1 (r = -0.230, p = .016) in NERD patients. In in vitro studies, expression of DPP10 in airway epithelial cells was enhanced by TGF-ß1 treatments. Furthermore, DPP10 was found to be produced from immune cells and this molecule induced the ERK phosphorylation in airway epithelial cells, which was suppressed by anti-DPP10 treatment. In asthmatic mouse models, increased levels of DPP10 in the serum and TGF-ß1 in the bronchoalveolar lavage fluid were noted, which were suppressed by anti-DPP10 treatment. Moreover, anti-DPP10 treatment inhibited the ERK phosphorylation and extracellular matrix deposition in the lungs. CONCLUSIONS AND CLINICAL RELEVANCE: These findings suggest that increased production of DPP10 may contribute to TGF-ß1-mediated airway dysfunction in NERD patients, where blockade of DPP10 may have potential benefits.
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Asma , Dipeptidil Peptidases e Tripeptidil Peptidases/metabolismo , Doenças Respiratórias , Animais , Anti-Inflamatórios não Esteroides , Asma/metabolismo , Dipeptidil Peptidases e Tripeptidil Peptidases/genética , Humanos , Pulmão/metabolismo , Camundongos , Doenças Respiratórias/patologia , Fator de Crescimento Transformador beta1RESUMO
Stratification of asthmatic patients based on relevant biomarkers enables the prediction of responsiveness against immune-targeted therapies in patients with asthma. Individualised therapy in patients with eosinophilic asthma has yielded improved clinical outcomes; similar approaches in patients with neutrophilic asthma have yet to be developed. We determined whether colony-stimulating factors (CSFs) in the airway reflect the inflammatory phenotypes of asthma and contribute to disease progression of neutrophilic asthma.We analysed three different mouse models of asthma and assessed cytokine profiles in sputum from human patients with asthma stratified according to inflammatory phenotype. In addition, we evaluated the therapeutic efficacy of various cytokine blockades in a mouse model of neutrophilic asthma.Among the CSFs, airway granulocyte CSF (G-CSF) contributes to airway neutrophilia by promoting neutrophil development in bone marrow and thereby distinguishes neutrophilic inflammation from eosinophilic inflammation in mouse models of asthma. G-CSF is produced by concurrent stimulation of the lung epithelium with interleukin (IL)-17A and tumour necrosis factor (TNF)-α; therefore, dual blockade of upstream stimuli using monoclonal antibodies or genetic deficiency of the cytokines in IL-17A×TNF-α double-knockout mice reduced the serum level of G-CSF, leading to alleviation of neutrophilic inflammation in the airway. In humans, the sputum level of G-CSF can be used to stratify patients with asthma with neutrophil-dominated inflammation.Our results indicated that myelopoiesis-promoting G-CSF and cytokines as the upstream inducing factors are potential diagnostic and therapeutic targets in patients with neutrophilic asthma.
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Asma , Asma/tratamento farmacológico , Humanos , Inflamação , Pulmão , Neutrófilos , EscarroRESUMO
BACKGROUND: Obesity associated with various complications has increased worldwide. Body weight gain alters lipid metabolites (especially sphingolipids) contributing to obesity-induced inflammation. However, the significance of the metabolites in the development of obese asthma is not yet clear. METHODS: The serum levels of sphingolipids were measured using liquid chromatography-tandem mass spectrometry in obese controls (n = 7) and patients with asthma: the obese group (BMI > 25 kg/m2 , n = 13) vs the nonobese (n = 28) group. To examine the relationship between metabolic changes in sphingolipids and macrophage polarization, public microarray data were analyzed. In addition, the alteration in sphingolipid metabolism was investigated in wild-type BALB/c mice fed a high-fat diet. RESULTS: The obese asthma had higher levels of serum C18:0 and C20:0 ceramides than the nonobese asthma group (P = .028 and P = .040, respectively). The value of the serum C18:0 ceramide (184.3 ng/mL) for discriminating the obese asthma from the nonobese asthma group showed 53.9% sensitivity and 85.7% specificity (AUC = 0.721, P = .024). The microarray data showed significantly increased ceramide synthesis and metabolic shift to ceramide accumulation during M1 macrophage polarization in humans. Increased airway hyperresponsiveness, M1 macrophage polarization, and C18:0 ceramide levels were noted in obese mice, but not in nonobese mice. Increased expression of ceramide synthase (CerS) 1 and CerS6 (not CerS2) was noted in lung tissues of obese mice. CONCLUSION: Alteration in sphingolipid metabolism favoring ceramide accumulation (especially long-chain ceramides) may contribute to developing obese asthma.
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Asma , Ceramidas , Animais , Asma/etiologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Obesidade/complicações , EsfingolipídeosRESUMO
BACKGROUND: Recent evidence demonstrates that activated eosinophils undergo a distinct form of lytic cell death, accompanied by formation of DNA-based eosinophil extracellular trap (EET) and degranulation, enhancing inflammatory immune responses in asthmatic airways. We previously showed that human blood eosinophils undergo degranulation in response to lysophosphatidylserine (LysoPS), an inflammatory lipid mediator, and strongly express P2Y10, a LysoPS receptor. METHODS: We evaluated EET, degranulation, and cell death of eosinophils in response to various concentrations of LysoPS. We also compared responsiveness to LysoPS between eosinophils from severe and nonsevere asthmatics. RESULTS: Extensive EET formation was elicited from a substantial fraction of stimulated eosinophils in response to 50 µmol/L LysoPS. Analyses for LDH and eosinophil-derived neurotoxin release showed that both lytic cell death and degranulation accompanied EET formation in response to LysoPS. Cytological analyses demonstrated that citrullinated histone 3 was present in the extracellular, filamentous DNA structure embedded with eosinophil granules. The LysoPS-induced EET was independent of ROS production and irrelevant to several signaling pathways examined, but dependent on protein arginine deiminase 4. A low concentration of LysoPS (5 µmol/L) did not induce EET or degranulation, but significantly increased platelet-activating factor-induced degranulation. Eosinophils from severe asthmatics exhibited greater degranulation, but not EET formation, in response to LysoPS (50 µmol/L), than those from nonsevere asthmatics, along with great expression of surface P2Y10. CONCLUSIONS: We identified a novel function of LysoPS, namely induction of EET in association with cytolysis and degranulation. LysoPS-dependent EET or degranulation plays a potential role in eosinophilic inflammation of severe asthma.
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Asma , Armadilhas Extracelulares , Degranulação Celular , Eosinófilos , Humanos , LisofosfolipídeosRESUMO
BACKGROUND: Activated eosinophils release extracellular traps (EETs), which contribute to airway inflammation in severe asthma (SA). However, the role of EETs in innate immunity has not yet been completely determined. The present study aimed to demonstrate the mechanism of airway inflammation in SA mediated by EETs. METHODS: Peripheral counts of EET+ eosinophils and type 2 innate lymphoid cells (ILC2s) were evaluated in patients with SA (n = 13), nonsevere asthma (NSA, n = 17), and healthy control subjects (HC, n = 8). To confirm the effect of EETs, airway hyperresponsiveness (AHR) and adapted/innate immune responses were assessed in mice. Furthermore, the effects of anti-IL-33/TSLP antibody were tested. RESULTS: The numbers of EET+ eosinophils and ILC2s were significantly elevated in SA, with a positive correlation between these two cells (r = .539, P < .001). When mice were injected with EETs, we observed significant increases in epithelium-derived cytokines (IL-1α, IL-1ß, CXCL-1, CCL24, IL-33, and TSLP) and eosinophil/neutrophil count in bronchoalveolar lavage fluid (BALF) as well as an increased proportion of IL-5- or IL-13-producing ILC2s in the lungs. When Rag1-/- mice receiving ILC2s were treated with EETs, increased AHR and IL-5/IL-13 levels in BALF were noted, which were effectively suppressed by anti-IL-33 or anti-TSLP antibody. CONCLUSION: EETs could enhance innate and type 2 immune responses in SA, in which epithelium-targeting biologics (anti-IL-33/TSLP antibody) may have a potential benefit.
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Asma/imunologia , Eosinófilos/imunologia , Armadilhas Extracelulares/imunologia , Linfócitos/imunologia , Mucosa Respiratória/imunologia , Adulto , Idoso , Animais , Feminino , Humanos , Subpopulações de Linfócitos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pessoa de Meia-IdadeRESUMO
PURPOSE OF REVIEW: Lung tissues are highly susceptible to airway inflammation as they are inevitably exposed to inhaled pathogens and allergens. In the lungs, clearance of infectious agents and regulation of inflammatory responses are important for the first-line defense, where surfactants play a role in host defense mechanisms. In this review, clinical significance of pulmonary surfactants in asthma has been highlighted. RECENT FINDINGS: Surfactants, such as surfactant protein A (SP-A) and SP-D released from alveolar epithelium, reduce pathogen infection and control immune-cell activation. Especially, SP-D directly binds to eosinophil surface, leading to inhibition of extracellular trap formation and reduction in airway inflammation. Production of surfactants is commonly determined by both genetic (single nucleotide polymorphisms) and environmental factors influencing processes involved in the development of asthma. In addition, nintedanib (an intracellular inhibitor of tyrosine kinases) could increase SP-D levels and is used in patients with idiopathic pulmonary fibrosis. These findings may provide a possible application of SP-D in asthma. Surfactants are key players contributing to host defense through maintaining the immune system. As clinical implications of surfactants involved in asthma have been suggested, further translational studies are needed to apply surfactants as an effective therapeutic target in patients with asthma.
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Asma/tratamento farmacológico , Surfactantes Pulmonares/uso terapêutico , Humanos , Surfactantes Pulmonares/farmacologiaRESUMO
Platelets modulate asthma pathogenesis by forming the platelet-eosinophil aggregation (PEA), which facilitates the activation of eosinophils. Platelets exhibit the purinergic receptor (P2Y12R), which responds to cysteinyl leukotriene E4 (LTE4 ). We have suggested that the combination of an antiplatelet drug (clopidogrel, [Clo]) and montelukast (Mon) would synergistically suppress asthma. BALB/c mice were intraperitoneally sensitized with ovalbumin (OVA) on days 0 and 14 and subsequently challenged on days 28-30 and 42-44. Mice were administered with Clo (10 mg/kg), Mon (10 mg/kg) or both drugs (Clo/Mon) orally 30 minutes before the OVA (1%) challenge on days 42-44. Mice were assayed for airway hyper-responsiveness (AHR) to methacholine and airway inflammation. Clopidogrel and montelukast attenuated the increased AHR; the combined treatment was more effective than a single treatment for total and eosinophil counts (all P < 0.05). Levels of interleukin (IL)-4, IL-5, IL-13, platelet factor 4, eosinophil peroxidase and LTE4 increased in the bronchoalveolar lavage fluid of asthmatic mice, but these levels decreased in mice treated with Clo/Mon (all P < 0.05). Goblet cell hyperplasia decreased in response to Clo/Mon. Mouse platelets and eosinophils were isolated and co-cultured for an in vitro assay with 10 µmol/L adenosine diphosphate (ADP), LTE4 (200 nmol/L), Mon (1 µmol/L), Clo (1 µmol/L) and Clo/Mon (1 µmol/L). Flow cytometry revealed that the increased formation of the PEA (%) was fully mediated by ADP and partly mediated by LTE4 . Clo/Mon reduced ADP-induced PEA formation and P-selectin expression (P < 0.05). In conclusion, Clo/Mon synergistically relieved asthma by inhibiting ADP-mediated PEA formation.
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Acetatos/uso terapêutico , Asma/tratamento farmacológico , Clopidogrel/uso terapêutico , Quinolinas/uso terapêutico , Difosfato de Adenosina/farmacologia , Animais , Asma/sangue , Asma/fisiopatologia , Biomarcadores/metabolismo , Líquido da Lavagem Broncoalveolar , Agregação Celular/efeitos dos fármacos , Quimiocinas/metabolismo , Ciclopropanos , Citocinas/metabolismo , Sinergismo Farmacológico , Eosinófilos/metabolismo , Eosinófilos/patologia , Feminino , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Contagem de Leucócitos , Leucotrieno E4/sangue , Leucotrieno E4/metabolismo , Pulmão/patologia , Camundongos Endogâmicos BALB C , Muco/metabolismo , Ativação Plaquetária/efeitos dos fármacos , Hipersensibilidade Respiratória/complicações , Hipersensibilidade Respiratória/fisiopatologia , Sulfetos , Células Th2/metabolismoRESUMO
Our previous study revealed that the ethanolic extract of Justicia procumbens ameliorates ovalbumin-induced airway inflammation and airway hyper-responsiveness in a mouse model of asthma. However, the mechanism of action of the extract remains unknown. In this study, we prepared DW2008S, an optimized and standardized powder extracted from J. procumbens using anhydrous ethanol, and investigated its anti-asthmatic effect and mechanism of action. Our results showed that DW2008S contains two major ingredients, justicidin A (JA) and justicidin B (JB), which selectively inhibit T helper 2 (Th2) cell responses in concanavalin A-activated spleen cells and polarized Th2 cells. Blockade of T cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibition motif domains (TIGIT) using a neutralizing antibody also selectively inhibited Th2 cell responses. Furthermore, DW2008S regulated TIGIT expression in the mice and cultured cells. Additionally, DW2008S and JA antagonized human adenosine receptor A3 (A3 AR), which mediates mast cell-dependent inflammation and bronchoconstriction. DW2008S and JB inhibited human phosphodiesterase 4 (PDE4), which is known to cause bronchoconstriction; however, the required concentrations were higher than those needed to affect TIGIT . These findings suggest that DW2008S can potentially ameliorate Th2-driven airway inflammation and bronchoconstriction through negative regulation of TIGIT and blockade of A3 AR and PDE4 activities.
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Antiasmáticos/uso terapêutico , Asma/tratamento farmacológico , Justicia/química , Extratos Vegetais/uso terapêutico , Animais , Antiasmáticos/farmacologia , Anticorpos Neutralizantes/farmacologia , Asma/patologia , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/metabolismo , Dioxolanos/farmacologia , Modelos Animais de Doenças , Feminino , Inflamação/imunologia , Inflamação/patologia , Lignanas/farmacologia , Camundongos Endogâmicos BALB C , Extratos Vegetais/farmacologia , Substâncias Protetoras/farmacologia , Substâncias Protetoras/uso terapêutico , Receptor A3 de Adenosina/metabolismo , Receptores Imunológicos/metabolismo , Células Th2/efeitos dos fármacos , Células Th2/imunologiaRESUMO
BACKGROUND & AIMS: Obesity and metabolic syndrome have been associated with alterations to the intestinal microbiota. However, few studies examined the effects of obesity on the intestinal immune system. We investigated changes in subsets of intestinal CD4+ T-helper (TH) cells with obesity and the effects of gut-tropic TH17 cells in mice on a high-fat diet (HFD). METHODS: We isolated immune cells from small intestine and adipose tissue of C57BL/6 mice fed a normal chow diet or a HFD for 10 weeks and analyzed the cells by flow cytometry. Mice fed a vitamin A-deficient HFD were compared with mice fed a vitamin A-sufficient HFD. Obese RAG1-deficient mice were given injections of only regulatory T cells or a combination of regulatory T cells and TH17 cells (wild type or deficient in integrin ß7 subunit or interleukin 17 [IL17]). Mice were examined for weight gain, fat mass, fatty liver, glucose tolerance, and insulin resistance. Fecal samples were collected before and after T cell transfer and analyzed for microbiota composition by metagenomic DNA sequencing and quantitative polymerase chain reaction. RESULTS: Mice placed on a HFD became obese, which affected the distribution of small intestinal CD4+ TH cells. Intestinal tissues from obese mice had significant reductions in the proportion of TH17 cells but increased proportion of TH1 cells, compared with intestinal tissues from nonobese mice. Depletion of vitamin A in obese mice further reduced the proportion of TH17 cells in small intestine; this reduction correlated with more weight gain and worsening of glucose intolerance and insulin resistance. Adoptive transfer of in vitro-differentiated gut-tropic TH17 cells to obese mice reduced these metabolic defects, which required the integrin ß7 subunit and IL17. Delivery of TH17 cells to intestines of mice led to expansion of commensal microbes associated with leanness. CONCLUSIONS: In mice, intestinal TH17 cells contribute to development of a microbiota that maintains metabolic homeostasis, via IL17. Gut-homing TH17 cells might be used to reduce metabolic disorders in obese individuals.
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Transferência Adotiva , Imunidade nas Mucosas , Resistência à Insulina , Intestino Delgado/imunologia , Síndrome Metabólica/prevenção & controle , Obesidade/prevenção & controle , Células Th17/transplante , Animais , Células Cultivadas , Dieta Hiperlipídica , Modelos Animais de Doenças , Fezes/microbiologia , Microbioma Gastrointestinal/imunologia , Genótipo , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Interações Hospedeiro-Patógeno , Cadeias beta de Integrinas/genética , Cadeias beta de Integrinas/metabolismo , Interleucina-17/deficiência , Interleucina-17/genética , Intestino Delgado/metabolismo , Intestino Delgado/microbiologia , Masculino , Síndrome Metabólica/genética , Síndrome Metabólica/imunologia , Síndrome Metabólica/microbiologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Obesidade/genética , Obesidade/imunologia , Obesidade/microbiologia , Fenótipo , Células Th17/imunologia , Células Th17/microbiologia , Fatores de Tempo , Deficiência de Vitamina A/complicaçõesRESUMO
BACKGROUND: Clinical features of aspirin-exacerbated respiratory disease (AERD) are characterized by overproduction of cysteinyl leukotrienes (LT) and eosinophil activation, in which epithelial cells contribute to eosinophilic airway inflammation. Folliculin (FLCN) helps maintain the integrity of epithelial barrier, but little is known about FLCN in AERD. OBJECTIVE: We investigated the role of FLCN in the pathogenic mechanisms of AERD. METHODS: We recruited 178 subjects with AERD, 276 subjects with aspirin-tolerant asthma (ATA) and 71 normal healthy controls (NC) at Ajou Medical Center. Levels of FLCN and interleukin (IL)-8 in sera and supernatants were measured by ELISA. Peripheral blood eosinophils isolated from asthmatic patients were cocultured with human airway epithelial cells (HAECs) pretreated with LTE4 , dexamethasone and montelukast. The intracellular expressions of FLCN, tight (TJ) (occludins, claudin-1) and adherens (AJ) junctions (E-cadherin) were analysed by Western blotting. shRNA was used to down-regulate FLCN (shFLCN) in HAECs. RESULTS: Serum FLCN levels were significantly higher in AERD group than in ATA and NC groups (all P < 0.001). The cut-off value of 56.6 pg/mL was used to define the high FLCN phenotype (highFLCN). Asthmatic patients with highFLCN were associated with increased airway hyperresponsiveness to methacholine (P = 0.015). The serum FLCN level could discriminate AERD group from NC group with 82% sensitivity (AUC = 0.793, P < 0.001). When HAECs were exposed to LTE4 , FLCN release was increased significantly (P < 0.05), which were amplified along with disruption of TJ and AJ expressions when HAECs were cocultured with eosinophils and LTE4 (all P < 0.05); these effects were suppressed by dexamethasone and montelukast. FLCN knockdown reduced IL-8 release and occludin expression from shFLCN HAECs. CONCLUSIONS: Our findings suggest that high LT and airway eosinophilia increased FLCN release from HAECs, which enhance epithelial activation and disruption. Modulation of FLCN may be a potential target for AERD.
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Aspirina/efeitos adversos , Estrona/metabolismo , Mucosa Respiratória/imunologia , Mucosa Respiratória/metabolismo , Doenças Respiratórias/etiologia , Doenças Respiratórias/metabolismo , Adulto , Asma Induzida por Aspirina , Biomarcadores , Linhagem Celular , Biologia Computacional/métodos , Citocinas/metabolismo , Eosinófilos/imunologia , Eosinófilos/metabolismo , Feminino , Técnicas de Silenciamento de Genes , Humanos , Inflamação/diagnóstico , Inflamação/etiologia , Inflamação/metabolismo , Junções Intercelulares/metabolismo , Masculino , Pessoa de Meia-Idade , Fenótipo , República da Coreia , Testes de Função Respiratória , Mucosa Respiratória/patologia , Doenças Respiratórias/diagnósticoRESUMO
Asthma is a common chronic disease with several variant phenotypes and endotypes. NSAID-exacerbated respiratory disease (NERD) is one such endotype characterized by asthma, chronic rhinosinusitis (CRS) with nasal polyps, and hypersensitivity to aspirin/cyclooxygenase-1 inhibitors. NERD is more associated with severe asthma than other asthma phenotypes. Regarding diagnosis, aspirin challenge tests via the oral or bronchial route are a standard diagnostic method; reliable in vitro diagnostic tests are not available. Recent studies have reported various biomarkers of phenotype, diagnosis, and prognosis. In this review, we summarized the known potential biomarkers of NERD that are distinct from those of aspirin-tolerant asthma. We also provided an overview of the different NERD subgroups.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Biomarcadores/sangue , Animais , Aspirina/uso terapêutico , Asma/sangue , Asma/tratamento farmacológico , Asma/imunologia , Asma Induzida por Aspirina/sangue , Asma Induzida por Aspirina/tratamento farmacológico , Asma Induzida por Aspirina/imunologia , Biomarcadores/metabolismo , Humanos , Transtornos Respiratórios/sangue , Transtornos Respiratórios/tratamento farmacológico , Transtornos Respiratórios/imunologiaRESUMO
The hypothesis of autoimmune involvement in asthma has received much recent interest. Autoantibodies, such as anti-cytokeratin (CK) 18, anti-CK19, and anti-α-enolase antibodies, react with self-antigens and are found at high levels in the sera of patients with severe asthma (SA). However, the mechanisms underlying autoantibody production in SA have not been fully determined. The present study was conducted to demonstrate that neutrophil extracellular DNA traps (NETs), cytotoxic molecules released from neutrophils, are a key player in the stimulation of airway epithelial cells (AECs) to produce autoantigens. This study showed that NETs significantly increased the intracellular expression of tissue transglutaminase (tTG) but did not affect that of CK18 in AECs. NETs induced the extracellular release of both tTG and CK18 in a concentration-dependent manner. Moreover, NETs directly degraded intracellular α-enolase into small fragments. However, antibodies against neutrophil elastase (NE) or myeloperoxidase (MPO) attenuated the effects of NETs on AECs. Furthermore, each NET isolated from healthy controls (HC), nonsevere asthma (NSA), and SA had different characteristics. Taken together, these findings suggest that AECs exposed to NETs may exhibit higher autoantigen production, especially in SA. Therefore, targeting of NETs may represent a new therapy for neutrophilic asthma with a high level of autoantigens.
Assuntos
Autoantígenos/metabolismo , Células Epiteliais/metabolismo , Armadilhas Extracelulares/metabolismo , Neutrófilos/metabolismo , Adulto , Células Cultivadas , Feminino , Proteínas de Ligação ao GTP/metabolismo , Humanos , Elastase de Leucócito/metabolismo , Masculino , Proteína 2 Glutamina gama-Glutamiltransferase , Transglutaminases/metabolismo , Adulto JovemAssuntos
Antibacterianos/farmacologia , Asma/microbiologia , Azitromicina/farmacologia , Cefixima/farmacologia , Microbioma Gastrointestinal/efeitos dos fármacos , Alérgenos/imunologia , Animais , Asma/imunologia , Asma/terapia , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Líquido da Lavagem Broncoalveolar/microbiologia , Citocinas/imunologia , Transplante de Microbiota Fecal , Fezes/microbiologia , Feminino , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Células Th2/efeitos dos fármacos , Células Th2/imunologiaRESUMO
[Purpose] The purpose of the present study was to investigate the effects of whole body vibration exercise in the horizontal direction on balance and fear of falling in the elderly. [Methods] This study was a case series of 17 elderly individuals. Participants performed whole body vibration exercise in the horizontal direction using a whole body vibration device for 15 minutes a day, 3 times a week, for 6 weeks. At baseline and after the 6-week intervention, balance was measured using the Berg Balance Scale and Timed Up and Go test, and fear of falling was assessed using the Falls Efficacy Scale. [Results] After the intervention, significant improvements from baseline values in the Berg Balance Scale, Timed Up and Go test, and Falls Efficacy Scale were observed in the study participants. [Conclusion] Elderly individuals who performed whole body vibration exercise in the horizontal direction showed significant improvements in balance and fear of falling. However, the observed benefits of whole body vibration exercise in the horizontal direction need to be confirmed by additional studies.