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Eol-1 cell lines induce EMT in BEAS-2B cells by increasing TGF-b1 expression. (A, B) Control and BEAS-2B cells co-cultured with EoL-1 cells. (C) BEAS-2B cells co-cultured with EoL-1 cells in the presence of anti-TGF-b1 neutralizing antibody. BEAS-2B cells lysates were subjected to RT-PCR analysis. The expression level of E-cadherin (D) and vimentin (E) mRNA was measured by RT-PCR and normalized to that of GAPDH mRNA. TGF-b1 (F) levels in the supernatant from each culture condition, and the ratio of p-Smad3 to Smad3 (G) in BEAS-2B cells as analyzed by densitometry. The scale bars indicate 50 mm. Data are expressed as means 6 SEM. *P,0.05. doi:10.1371/journal.pone.0064281.g007
Source publication
Eosinophilic inflammation and remodeling of the airways including subepithelial fibrosis and myofibroblast hyperplasia are characteristic pathological findings of bronchial asthma. Epithelial to mesenchymal transition (EMT) plays a critical role in airway remodelling. In this study, we hypothesized that infiltrating eosinophils promote airway remod...
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Context 1
... primary human eosinophils, the human eosinophilic leukemia cell line, EoL-1 cells, also induced EMT in BEAS-2B cells ( Figure 7A, B). EMT changes in BEAS-2B cells induced by Eol-1 were also inhibited in the presence of anti-TGF-b1 mAb ( Figure 7C). ...
Context 2
... primary human eosinophils, the human eosinophilic leukemia cell line, EoL-1 cells, also induced EMT in BEAS-2B cells ( Figure 7A, B). EMT changes in BEAS-2B cells induced by Eol-1 were also inhibited in the presence of anti-TGF-b1 mAb ( Figure 7C). Changes in the mRNA expression of E-cadherin and vimentin observed in BEAS-2B cells co-cultured with Eol-1 were similar to those observed when the epithelial cell line was cultured with primary human eosinophils ( Figure 7D, E). ...
Context 3
... changes in BEAS-2B cells induced by Eol-1 were also inhibited in the presence of anti-TGF-b1 mAb ( Figure 7C). Changes in the mRNA expression of E-cadherin and vimentin observed in BEAS-2B cells co-cultured with Eol-1 were similar to those observed when the epithelial cell line was cultured with primary human eosinophils ( Figure 7D, E). Further, the protein level of TGF-b1 was significantly increased in the co- culture supernatant ( Figure 7F) and the p-Smad3 to Smad3 ratio was significantly high in BEAS-2B cells cultured with Eol-1 cells ( Figure 7G). ...
Context 4
... in the mRNA expression of E-cadherin and vimentin observed in BEAS-2B cells co-cultured with Eol-1 were similar to those observed when the epithelial cell line was cultured with primary human eosinophils ( Figure 7D, E). Further, the protein level of TGF-b1 was significantly increased in the co- culture supernatant ( Figure 7F) and the p-Smad3 to Smad3 ratio was significantly high in BEAS-2B cells cultured with Eol-1 cells ( Figure 7G). The need of cell-to-cell contact for induction of EMT in BEAS-2B by EoL-1 cells was also appraised using a trans- well system. ...
Context 5
... in the mRNA expression of E-cadherin and vimentin observed in BEAS-2B cells co-cultured with Eol-1 were similar to those observed when the epithelial cell line was cultured with primary human eosinophils ( Figure 7D, E). Further, the protein level of TGF-b1 was significantly increased in the co- culture supernatant ( Figure 7F) and the p-Smad3 to Smad3 ratio was significantly high in BEAS-2B cells cultured with Eol-1 cells ( Figure 7G). The need of cell-to-cell contact for induction of EMT in BEAS-2B by EoL-1 cells was also appraised using a trans- well system. ...
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Background:
Airway inflammation is an important characteristic of asthma and has been associated with airway remodelling and bronchial hyperreactivity. The mucosal microenvironment composed of structural cells and highly specialised extracellular matrix is able to amplify and promote inflammation. This microenvironment leads to the development and...
Background
The collagen gel contraction assay measures gel size to assess the contraction of cells embedded in collagen gel matrices. Using the assay with lung fibroblasts is useful in studying the lung tissue remodeling process in wound healing and disease development. However, the involvement of bronchial epithelial cells in this process should a...
Citations
... Epithelial plasticity is a mechanism that has been observed in asthmatic airways [15]. In co-culture studies, eosinophils, purified from human blood or cells of a butyrate-differentiated eosinophilic leukemic cell line (EoL-1), promoted epithelial plasticity in BEAS-2B cells, a virally transformed human epithelial cell line isolated from normal human bronchial epithelium, as evaluated by decreased E-cadherin (cadherin-1) mRNA and protein, and increased vimentin, SMAD3 phosphorylation, and transforming growth factor β1 (TGFB1) secretion [25]. Anti-TGFB1 inhibited the plasticity, which together with other inhibitor experiments indicated that TGFB1 (which is made by and present in eosinophils [26,27]) is responsible for or involved in the eosinophil effect by activating the JNK/SMAD3 pathway in epithelial cells. ...
... Anti-TGFB1 inhibited the plasticity, which together with other inhibitor experiments indicated that TGFB1 (which is made by and present in eosinophils [26,27]) is responsible for or involved in the eosinophil effect by activating the JNK/SMAD3 pathway in epithelial cells. In parallel mouse experiments, intratracheal instillation of mouse bone marrow-derived eosinophils led to decreased cadherin-1 and increased αsmooth muscle actin (ACTA2) staining [25], indicating that eosinophils can promote epithelial plasticity also in vivo. In a follow-up article by the same group, it was demonstrated that the β 2 adrenergic agonist procaterol inhibited epithelial plasticity in their co-culture system, supporting a beneficial effect of β 2 agonist on airway remodeling in asthma [28]. ...
Background: Eosinophils have numerous roles in type 2 inflammation depending on their activation states in the blood and airway or after encounter with inflammatory mediators. Airway epithelial cells have a sentinel role in the lung and, by instructing eosinophils, likely have a foundational role in asthma pathogenesis. Summary: In this review, we discuss various topics related to eosinophil-epithelial cell interactions in asthma, including the influence of eosinophils and eosinophil products, e.g., granule proteins, on epithelial cell function, expression, secretion, and plasticity; the effects of epithelial released factors, including oxylipins, cytokines, and other mediators on eosinophils, e.g., on their activation, expression, and survival; possible mechanisms of eosinophil-epithelial cell adhesion; and the role of intra-epithelial eosinophils in asthma. Key Messages: We suggest that eosinophils and their products can have both injurious and beneficial effects on airway epithelial cells in asthma and that there are bidirectional interactions and signaling between eosinophils and airway epithelial cells in asthma.
... Moreover, eosinophils play important roles in the development of airway remodeling in asthma [20][21][22]. For example, Humbles et al. reported that mice with selective ablation of the eosinophil lineage do not develop collagen deposition or increases in the airway smooth muscle cells in response to allergens, indicating the critical role of eosinophils in the development of airway remodeling [20]. ...
... For example, Humbles et al. reported that mice with selective ablation of the eosinophil lineage do not develop collagen deposition or increases in the airway smooth muscle cells in response to allergens, indicating the critical role of eosinophils in the development of airway remodeling [20]. Human eosinophils can induce the epithelialmesenchymal transition through transforming growth factor (TGF)-β1 [21]. We reported that in vivo IL-5 gene delivery induces eosinophilia and enhances the expression of TGF-β, whose signal is transduced to other cells [22]. ...
... A recent study has reported that dupilumab reduces airway eosinophilia despite a modest increase in the blood eosinophil count [46]. Eosinophils have been shown to play important roles in airway remodeling [20][21][22]. Furthermore, dupilumab inhibits the effects of IL-13 and IL-4 on human airway smooth muscle cells in vitro [55]. ...
Bronchial asthma is characterized by airway inflammation, airway hyperresponsiveness, and reversible airway obstruction. Eosinophils contribute to the pathogenesis of airway disease mainly by releasing eosinophil-specific granules, lipid mediators, superoxide anions, and their DNA. Type-2 cytokines such as interleukin (IL)-4 and IL-13 also play roles in the development of bronchial asthma. Among these cytokines, IL-4 is involved in T-cell differentiation, B-cell activation, B-cell differentiation into plasma cells, and the production of immunoglobulin E. Although IL-13 has similar effects to IL-4, IL-13 mainly affects structural cells, such as epithelial cells, smooth muscle cells, and fibroblasts. IL-13 induces the differentiation of goblet cells that produce mucus and induces the airway remodeling, including smooth muscle hypertrophy. IL-4 and IL-13 do not directly activate the effector functions of eosinophils; however, they can induce eosinophilic airway inflammation by upregulating the expression of vascular cell adhesion molecule-1 (for adhesion) and CC chemokine receptor 3 ligands (for migration). Dupilumab, a human anti-IL-4 receptor α monoclonal antibody that inhibits IL-4 and IL-13 signaling, decreases asthma exacerbations and mucus plugs and increases lung function in moderate to severe asthma. In addition, dupilumab is effective for chronic rhinosinusitis with nasal polyps and for atopic dermatitis, and IL-4/IL-13 blocking is expected to suppress allergen sensitization, including transcutaneous sensitization and atopic march.
... It hampers its regenerative abilities, leading to remodeling characterized by basement membrane thickening, mucous cell metaplasia, and the proliferation of smooth muscle cells and lung fibroblasts, among other changes. The exact mechanisms of chronic allergic airway inflammatory disorders remain unclear; however, it is believed that the EMT is involved in the progression from bronchial epithelial repair to remodeling in individuals with asthma [10,11]. The EMT is vital in embryonic development, tissue remodeling, and wound repair [12]. ...
... This transition enables enhanced motility in the cells [13,14]. Transforming growth factor beta 1 (TGF-β1), a multifunctional cytokine belonging to the TGF superfamily, is upregulated in airway remodeling tissues [10,11,15]. Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) and its receptor fibroblast growth factor-inducible 14 (Fn14) have emerged in the tumor necrosis factor (TNF) superfamily, promoting NF-κB activation, and the TWEAK/Fn14 interaction contributes to pro-inflammatory effects and the EMT process in the bronchial epithelium [6,[16][17][18]. ...
The mitogen-activated protein kinase (MAPK) signaling pathway is involved in the epithelial–mesenchymal transition (EMT) and asthma; however, the role of mitogen-activated protein kinase kinase kinase 19 (MAP3K19) remains uncertain. Therefore, we investigated the involvement of MAP3K19 in in vitro EMT and ovalbumin (OVA)-induced asthma murine models. The involvement of MAP3K19 in the EMT and the production of cytokines and chemokines were analyzed using a cultured bronchial epithelial cell line, BEAS-2B, in which MAP3K19 was knocked down using small interfering RNA. We also evaluated the involvement of MAP3K19 in the OVA-induced asthma murine model using Map3k19-deficient (MAP3K19−/−) mice. Transforming growth factor beta 1 (TGF-β1) and tumor necrosis factor-like weak inducer of apoptosis (TWEAK) induced the MAP3K19 messenger RNA (mRNA) expression in the BEAS-2B cells. The knockdown of MAP3K19 enhanced the reduction in E-cadherin mRNA and the production of regulated upon activation normal T cell express sequence (RANTES) via stimulation with TWEAK alone or with the combination of TGF-β1 and TWEAK. Furthermore, the expression of MAP3K19 mRNA was upregulated in both the lungs and tracheas of the mice in the OVA-induced asthma murine model. The MAP3K19−/− mice exhibited worsened eosinophilic inflammation and an increased production of RANTES in the airway epithelium compared with the wild-type mice. These findings indicate that MAP3K19 suppressed the TWEAK-stimulated airway epithelial response, including adhesion factor attenuation and RANTES production, and suppressed allergic airway inflammation in an asthma mouse model, suggesting that MAP3K19 regulates allergic airway inflammation in patients with asthma.
... As already highlighted, GFs, such as TGF-β1, are also involved in EMT induced by inflammatory cells, and it has been reported in several studies that TGF-β1 expression correlates with the number of eosinophils and the degree of airway remodelling in bronchial asthma [96,97]. To evaluate the effect of eosinophils on EMT, Yasukawa et al. [98] carried out in vitro and in vivo studies. In particular, they assessed EMT in mice instilled with bone-marrow-derived eosinophils and inhuman bronchial epithelial cells (BEC) co-cultured with eosinophils. ...
Epithelial-to-mesenchymal transition (EMT) is a reversible process, in which epithelial cells lose their epithelial traits and acquire a mesenchymal phenotype. This transformation has been described in different lung diseases, such as lung cancer, interstitial lung diseases, asthma, chronic obstructive pulmonary disease and other muco-obstructive lung diseases, such as cystic fibrosis and non-cystic fibrosis bronchiectasis. The exaggerated chronic inflammation typical of these pulmonary diseases can induce molecular reprogramming with subsequent self-sustaining aberrant and excessive profibrotic tissue repair. Over time this process leads to structural changes with progressive organ dysfunction and lung function impairment. Although having common signalling pathways, specific triggers and regulation mechanisms might be present in each disease. This review aims to describe the various mechanisms associated with fibrotic changes and airway remodelling involved in chronic airway diseases. Having better knowledge of the mechanisms underlying the EMT process may help us to identify specific targets and thus lead to the development of novel therapeutic strategies to prevent or limit the onset of irreversible structural changes.
... In experiments using nasal polyp-derived fibroblasts, fibroblasts stimulated with TLRs ligand secrete MCP-4, an eosinophil chemoattractant protein. This suggests that fibroblasts contribute to the inflammatory response in Th2 type or eosinophilic CRS [76,102,103]. Eosinophils are granular innate immune cells that play important roles in type 2 inflammation. Eosinophilia in the upper respiratory tract is associated with tissue remodeling and inflammation and induces EMT in bronchial cells to transform epithelial cells into mesenchymal cells. ...
Chronic rhinosinusitis (CRS) is a multifactorial inflammatory disease of the nose and sinuses that affects more than 10% of the adult population worldwide. Currently, CRS is classified into endotypes according to the inflammatory response (Th1, Th2, and Th17) or the distribution of immune cells in the mucosa (eosinophilic and non-eosinophilic). CRS induces mucosal tissue remodeling. Extracellular matrix (ECM) accumulation, fibrin deposition, edema, immune cell infiltration, and angiogenesis are observed in the stromal region. Conversely, epithelial-to-mesenchymal transition (EMT), goblet cell hyperplasia, and increased epithelial permeability, hyperplasia, and metaplasia are found in the epithelium. Fibroblasts synthesize collagen and ECM, which create a structural skeleton of tissue and play an important role in the wound-healing process. This review discusses recent knowledge regarding the modulation of tissue remodeling by nasal fibroblasts in CRS.
... It was also reported that eosinophil TLR receptors (TLR3, TLR4, TLR7, TLR8, and TLR9) were expressed in differentiated EOL-1 cells. In in vitro studies, primary eosinophils have disadvantages such as low survival rate and difficult long-term co-culture, so the EOL-1 cell line is often used as an alternative cell (13)(14)(15). ...
... These eosinophils are also involved in regulating other cells around them (25). Eosinophils induce the epithelial-to-mesenchymal transition in bronchial cells, leading to remodeling of the epithelium (15,26). Eosinophils control migration and infiltration of immune cells by regulating the surface marker expression in endothelial cells (27,28). ...
Background
Eosinophilic chronic rhinosinusitis (ECRS) is a subtype of chronic rhinosinusitis (CRS) and is a refractory or intractable disease. However, a reliable clinical marker or an effective treatment strategy has not yet been established. ECRS is accompanied by excessive eosinophil infiltration and Th2 inflammatory response, which is closely related to tissue remodeling in the upper airways.
Objectives
We sought to investigate the effect of eosinophils on tissue remodeling in ECRS. The purpose of this study was to identify the effects of eosinophils on the expression of pro-inflammatory mediators and extracellular matrix (ECM) in nasal fibroblasts and the key mediators that stimulate them.
Methods
Butyric acid was used to differentiate EOL-1 cells into eosinophils. We co-cultured differentiated EOL-1 cells and fibroblasts to measure the expression of pro-inflammatory mediators and ECM in fibroblasts. Among the cytokines secreted from the differentiated EOL-1 cells, factors that induced tissue remodeling of fibroblasts were identified.
Results
Treatment with butyric acid (BA) differentiated EOL-1 cells into eosinophils. Differentiated EOL-1 cells induced fibroblasts to produce pro-inflammatory mediators, IL-6 and IL-8, and tissue remodeling factor, VEGF. It also induced myofibroblast differentiation and overexpression of ECM components. Differentiated EOL-1 cells overexpressed osteopontin (OPN), and recombinant OPN increased the expression of IL-6, IL-8, VEGF, and ECM components in nasal fibroblast. OPN was overexpressed in the nasal tissue of patients with ECRS and was associated with the severity of CRS.
Conclusions
Eosinophil-derived OPN stimulated nasal fibroblasts and contributed to inflammation and tissue remodeling in ECRS. Moreover, the expression level of OPN was proportional to the severity of ECRS. Therefore, OPN regulation is a potential treatment for ECRS.
... MRGPRX2 is upregulated in asthmatic lungs [57]. Therefore, it is likely that hemokinin-1 (HK-1) produced from lung macrophages, bronchial cells and mast cells [59,60], as well as ligands generated from eosinophils [61,62], neutrophils [63], and epithelial cells [64], further activate mast cells via MRGPRX2 to release inflammatory mediators and thereby amplify airway hyper-responsiveness, resulting in allergic asthma exacerbation [65]. Furthermore, Perner et al. revealed that allergen-induced SP release by transient receptor potential vanilloid (TRPV) + sensory neurons triggers Masrelated G-protein coupled receptor member A1 (MRGPRA-1)-dependent DC migration to the lymph node, initiating the allergic immune response [66]. ...
Chronic obstructive airway diseases are characterized by airflow obstruction and airflow limitation as well as chronic airway inflammation. Especially bronchial asthma and chronic obstructive pulmonary disease (COPD) cause considerable morbidity and mortality worldwide, can be difficult to treat, and ultimately lack cures. While there are substantial knowledge gaps with respect to disease pathophysiology, our awareness of the role of neurological and neuro-immunological processes in the development of symptoms, the progression, and the outcome of these chronic obstructive respiratory diseases, is growing. Likewise, the role of pathogenic and colonizing microorganisms of the respiratory tract in the development and manifestation of asthma and COPD is increasingly appreciated. However, their role remains poorly understood with respect to the underlying mechanisms. Common bacteria and viruses causing respiratory infections and exacerbations of chronic obstructive respiratory diseases have also been implicated to affect the local neuro-immune crosstalk. In this review, we provide an overview of previously described neuro-immune interactions in asthma, COPD, and respiratory infections that support the hypothesis of a neuro-immunological component in the interplay between chronic obstructive respiratory diseases, respiratory infections, and respiratory microbial colonization.
... 164,165 In vitro evidence also indicates secretion of fibroproliferative and fibrogenic growth factors including FGF-2, insulin-like growth factors 1 (IGF-1), PDGF, ET-1, and TGF-β, which can induce collagen gene expression. [166][167][168][169] It was observed that eosinophil and epithelial cell interaction causes the release of TGF-β1 by bronchial epithelial cells, which activates JNK/Smad3 pathway causing EMT of airway epithelial cells. 170 EMT plays a significant role in airway remodeling. ...
Eosinophils are the major inflammatory cells which play a crucial role in the development of allergic and non-allergic asthma phenotypes. Eosinophilic asthma is the most heterogeneous phenotype where activated eosinophils are reported to be significantly associated with asthma severity. Activated eosinophils display an array of cell adhesion molecules that not only act as an activation marker, suitable for assessing severity, but also secrete several tissue factors, cytokines and chemokines which modulate the clinical severity. Eosinophil activations are also strictly associated with activation of other hetero cellular populations like neutro-phils, macrophages, mast cells, and platelets which culminate in the onset and progression of abnormal phenotypes such as bronchoconstriction, allergic response, fibrosis instigated by tissue inflammation, epithelial injury, and oxidative stress. During the activated state, eosino-phils release several potent toxic signaling molecules such as major basic proteins, eosinophil peroxidase, eosinophil cationic protein (ECP), and lipid mediators, rendering tissue damage and subsequently leading to allergic manifestation. The tissue mediators render a more complex manifestation of a severe phenotype by activating prominent signaling cross-talk. Here, in the current review with the help of search engines of PubMed, Medline, etc, we have tried to shed light and explore some of the potent determinants regulating eosinophil activation leading to asthma phenotype.
... viruses, bacteria, parasites, toxic environmental particles), a series of immunity activating signals are produced, leading to inflammation and promoting EMT. Immune cells, such as macrophages, neutrophils, and eosinophils, are recruited in this process and release cytokines and growth factors to maintain inflammation and tissue repair [227][228][229][230][231][232]. However, when this process is sustained, the development of a chronic inflammatory state enhances and prolongs EMT through increased fibroblast proliferation. ...
Idiopathic pulmonary fibrosis (IPF), the most common form of idiopathic interstitial pneumonia, is a progressive, irreversible, and typically lethal disease characterized by an abnormal fibrotic response involving vast areas of the lungs. Given the poor knowledge of the mechanisms underpinning IPF onset and progression, a better understanding of the cellular processes and molecular pathways involved is essential for the development of effective therapies, currently lacking. Besides a number of established IPF-associated risk factors, such as cigarette smoking, environmental factors, comorbidities, and viral infections, several other processes have been linked with this devastating disease. Apoptosis, senescence, epithelial-mesenchymal transition , endothelial-mesenchymal transition, and epithelial cell migration have been shown to play a key role in IPF-associated tissue remodeling. Moreover, molecules, such as chemokines, cytokines, growth factors, adenosine, glycosaminoglycans, non-coding RNAs, and cellular processes including oxidative stress, mitochondrial dysfunction, endoplasmic reticulum stress, hypoxia, and alternative polyadenylation have been linked with IPF development. Importantly, strategies targeting these processes have been investigated to modulate abnormal cellular phenotypes and maintain tissue homeostasis in the lung. This review provides an update regarding the emerging cellular and molecular mechanisms involved in the onset and progression of IPF.
... In addition to mast cells, granulocytes such as eosinophils and neutrophils also play a role in asthma pathogenesis. Upon activation, eosinophils release cytoplasmic proteins such as eosinophil-derived major basic protein (MBP) and eosinophil peroxidase (EPO) [114][115][116]. Lung neutrophils are also shown to secrete cathelicidin LL-37 in asthmatic patients Fig. 1 MRGPRX2-mediated mast cell-nociceptor interaction in atopic dermatitis, itch, and allergic asthma. ...
Purpose of Review
Atopic dermatitis (AD) and allergic asthma are complex disorders with significant public health burden. This review provides an overview of the recent developments on Mas-related G protein-coupled receptor-X2 (MRGPRX2; mouse counterpart MrgprB2) as a potential candidate to target neuro-immune interaction in AD and allergic asthma.
Recent Findings
Domestic allergens directly activate sensory neurons to release substance P (SP), which induces mast cell degranulation via MrgprB2 and drives type 2 skin inflammation in AD. MRGPRX2 expression is upregulated in human lung mast cells and serum of asthmatic patients. Both SP and hemokinin-1 (HK-1 generated from macrophages, bronchial cells, and mast cells) cause degranulation of human mast cells via MRGPRX2.
Summary
MrgprB2 contributes to mast cell-nerve interaction in the pathogenesis of AD. Furthermore, asthma severity is associated with increased MRGPRX2 expression in mast cells. Thus, MRGPRX2 could serve as a novel target for modulating AD and asthma.
