thromboplastin and Asthma

The number of single nucleotide polymorphisms (SNPs) found to be associated with asthma and related phenotypes outnumbers those with functional impacts. In this review we briefly described some of the approaches used to investigate functionality of SNPs, and summarized recent findings related to the characterization of functional SNPs in asthma.. For disease-associated SNPs residing in the promoter or 3' untranslated regions, differential protein binding affinity between the major and minor alleles is often the first logical area of investigation. In this review, we described SNPs associated with asthma or related phenotypes in five genes which in the past 12 months have new data implicating potential mechanisms in asthma development.. Variability in treatment responses poses a great challenge in asthma management. It is established that the genetic makeup of individuals plays a role in asthma development, yet the mechanisms remain unclear. Investigations on the functional impacts of disease-associated SNPs will help us gain insights into potential disease mechanisms, and ultimately lead to effective therapies for those who suffer from asthma.

Topics: Alleles; Asthma; DNA-Binding Proteins; Genetic Association Studies; Genetic Predisposition to Disease; Haplotypes; Humans; Membrane Proteins; Phenotype; Polymorphism, Single Nucleotide; Protein Binding; Receptors, Calcitriol; Receptors, Lysosphingolipid; Sphingosine-1-Phosphate Receptors; Thromboplastin; Transcription Factors

To detect the expressions of tumor necrosis factor-α-induced protein-8-like 2 (TIPE2) and tissue factor (TF) in peripheral blood mononuclear cells (PBMCs) of patients with acute exacerbation of bronchial asthma (BA), and to analyze their associations with changes in inflammatory factors and T lymphocytes.. A total of 59 patients with BA treated in our hospital from February 2018 to April 2019 were selected as objects, including 30 cases in the acute exacerbation phase (BA group) and 29 in the remission phase (RE group). During the same period, 28 people receiving physical examinations were selected as healthy controls (Control group). The proportion of eosinophils in the sputum and the fractional exhaled nitric oxide (FeNO) level were detected in each subject. Blood samples were collected in patients of BA group and Control group, aiming to isolate PBMCs. Then, the messenger RNA (mRNA) expressions of TIPE2 and TF in PBMCs were determined via reverse transcription-polymerase chain reaction (RT-PCR). Serum levels of interleukin-1β (IL-1β) and IL-6 in BA group and Control group were determined via enzyme-linked immunosorbent assay (ELISA), and protein expressions of serum T helper 1 (Th1) and Th2 cells in BA group and Control group were detected using Western blotting.. Compared with those in Control group, the proportion of eosinophils and FeNO level increased in BA and RE group, which were more pronounced in BA group. Downregulated mRNA level of TIPE2, and upregulated TF were detected in BA and RE groups compared to those of Control group, and the expression changes were more significant in the former group. Enzyme-linked immunosorbent assay (ELISA) data showed that serum levels of IL-1β and IL-6 were significantly elevated in BA and RE groups in comparison to Control group, especially BA group. In addition, protein level of Th1 cells was downregulated, while that of Th2 was upregulated in BA group and RE group compared to those of Control group, and a more significant change was observed in BA group compared to that of RE group.. In patients with acute exacerbation of BA, the expression of TIPE2 in PBMCs declined, while that of TF rose, which were negatively correlated with each other. Moreover, the proportion of Th1 cells declined in patients with acute exacerbation of BA, indicating that it is associated with the lung function, inflammatory level and proportion of eosinophils.

Topics: Acute Disease; Adult; Aged; Asthma; Female; Humans; Interleukin-1beta; Interleukin-6; Intracellular Signaling Peptides and Proteins; Leukocytes, Mononuclear; Male; Middle Aged; Th1 Cells; Thromboplastin

It has recently been shown that expression levels of tissue factor (TF) are high in the serum and peripheral blood mononuclear cells of patients with asthma. However, whether TF impacts airway inflammation and remodelling in asthma remains unknown. The aim of this study was to investigate the effect of TF in asthma airway inflammation and remodelling using a house dust mite (HDM)-induced chronic asthma model and human bronchial epithelial (16HBE) cells. A chronic asthma model was constructed in BALB/c mice by the intranasal instillation of HDM. Mice were treated with short hairpin TF (shTF), and airway inflammation and remodelling features of asthma and epithelial-mesenchymal transition (EMT) were assessed. 16HBE cells were induced by transforming growth factor-β1 (TGF-β1) and HDM in the presence or absence of shTF; then, EMT markers and invasion and migration ability were determined. TF expression increased in the lung tissue and 16HBE cells when exposed to HDM. TF downregulation in the lung significantly reduced airway hyperresponsiveness, eosinophil inflammation, the EMT process, and levels of interleukin (IL)-4, IL-6, IL-13, and TGF-β1 in bronchoalveolar lavage fluid of asthmatic mice. Moreover, TF downregulation inhibited migration and incursion and decreased the expression levels of fibronectin 1 and TGF-β1, but increased the expression of E-cadherin in HDM- and TGF-β1-stimulated 16HBE cells. These results demonstrated that TF promoted airway pathological features by enhancing the EMT of bronchial epithelial cells both in vitro and in mice with house dust mite-induced asthma.

Topics: Airway Remodeling; Allergens; Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Dermatophagoides pteronyssinus; Disease Models, Animal; Epithelial Cells; Epithelial-Mesenchymal Transition; HEK293 Cells; Humans; Mice; Specific Pathogen-Free Organisms; Thromboplastin; Up-Regulation

Tissue factor (TF), a primary initiator of blood coagulation, also plays a pivotal role in angiogenesis. TF expression in the airways is associated with asthma, a disease characterized in part by subepithelial angiogenesis.. To determine potential sources of TF and the mechanisms of its availability in the lung microenvironment.. Normal human bronchial epithelial cells grown in air-liquid interface culture were subjected to a compressive stress of 30 cm H(2)O; this is comparable to that generated in the airway epithelium during bronchoconstriction in asthma. Conditioned media and cells were harvested to measure TF mRNA and TF protein. We also tested bronchoalveolar lavage fluid and airway biopsies from asthmatic patients and healthy controls for TF.. TF mRNA was upregulated 2.2-fold after 3 hours of stress compared with unstressed cells. Intracellular and secreted TF proteins were enhanced 1.6-fold and more than 50-fold, respectively, compared with those of control cells after the onset of compression. The amount of TF in the bronchoalveolar lavage fluid from patients with asthma was found at mean concentrations that were 5 times greater than those of healthy controls. Immunohistochemical staining of endobronchial biopsies identified epithelial localization of TF with increased expression in asthma. Exosomes isolated from the conditioned media of normal human bronchial epithelial cells and the bronchoalveolar lavage fluid of asthmatic subjects by ultracentrifugation contained TF.. Our in vitro and in vivo studies show that mechanically stressed bronchial epithelial cells are a source of secreted TF and that exosomes are potentially a key carrier of the TF signal.

Topics: Adult; Aged; Airway Remodeling; Asthma; Biopsy; Bronchi; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Cells, Cultured; Cellular Microenvironment; Epithelial Cells; Exosomes; Humans; Mechanotransduction, Cellular; Middle Aged; Thromboplastin; Young Adult

Tissue factor (TF) is important for initiation of coagulation and for the increased thrombin activity observed at sites of inflammation. Thrombin activity is induced by allergen challenge in asthmatic airways and is involved in the pathogenesis of asthma. A -603A --> G polymorphism (rs1361600) in the promoter region of the TF gene has been associated with serum TF levels and with the development of cardiovascular diseases. The aim of this study was to determine whether the functional -603A --> G polymorphism has genetic influences on the development of asthma. Case-control analysis was performed of the association between six common single-nucleotide polymorphisms (SNPs), including the -603A --> G polymorphism, at the TF gene, and the development of asthma, using two unrelated Japanese populations. In the primary population (n=826), the GG genotype at the -603A --> G polymorphism was associated with adult-onset asthma (onset at >or=21 years of age) (odds ratio (OR) 2.886, P=0.0231). A second population showed a similar tendency (n=1654, OR 1.602, P=0.064). Transcriptional activity of promoters with -603A --> G genotypes were examined using luciferase promoter assays. The -603G allele was associated with higher promoter activity (P<0.05). The association between the functional polymorphism (-603A --> G) in the TF gene promoter and adult-onset asthma indicates that TF is a candidate gene contributing to asthma susceptibility.

Topics: Adolescent; Adult; Age of Onset; Aged; Alleles; Asthma; Child; Child, Preschool; Female; Genetic Predisposition to Disease; Humans; Infant; Japan; Kaplan-Meier Estimate; Male; Middle Aged; Nuclear Proteins; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Protein Binding; Reproducibility of Results; Thromboplastin; Transcription, Genetic; Young Adult

Eosinophil counts in the bronchoalveolar lavage fluid of wild-type (WT) mice increased after ovalbumin (OVA) challenge, a response that was diminished in comparably challenged low-expressing coagulation factor VII (FVII(tTA/tTA)) mice. Levels of T helper type 2 (Th2) cytokines, IL-4, IL-5, and IL-13, and eosinophil-attracting chemokines, eotaxin and RANTES, were also lower in the OVA-challenged FVII(tTA/tTA) mice. Eosinophils purified from low-FVII mice underwent apoptosis at a faster rate compared with WT eosinophils, and eosinophil migration in response to eotaxin was reduced in eosinophils obtained from FVII(tTA/tTA) mice. Airway hyperresponsiveness and mucous layer thickness were reduced in OVA-treated FVII(tTA/tTA) mice, and addition of exogenous coagulation factor X (FX) enhanced mucin production in human epithelial NCI-H292 cells. Correspondingly, incubation of FX with NCI-H292 cells resulted in activated (a) FX production, suggesting that the components required for FX activation were present on NCI-H292 cells. These results demonstrate that FVIIa functions in the asthmatic response to an allergen by stimulating lung eosinophilia, airway hyperresponsiveness, and mucin production, this latter effect through its ability to activate FX in conjunction with tissue factor.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Count; Cell Line; Cell Movement; Cell Survival; Cytokines; Eosinophils; Epithelial Cells; Factor VIIa; Female; Gene Expression Regulation; Inflammation; Inflammation Mediators; Lung; Mice; Mice, Inbred C57BL; Mucins; Ovalbumin; RNA, Messenger; Thromboplastin

Allergic asthma is characterized by bronchial inflammation and repair processes at the same time, that cause increased airway obstruction. Recent evidences suggest that monocytes and macrophages may play important role in allergic inflammation. Activated by proinflammatory factors they can express tissue factor (TF) and tissue factor pathway inhibitor (TFPI). The aim of the present study was to evaluate TF and TFPI in plasma of bronchial asthma patients. The study was performed on 17 asthma patients with documented Dermatophagoides pteronyssinus (Dp) allergy. 15 healthy persons served as negative control. Specific bronchial challenge with Dp extracts was performed in asthma patients. Blood was collected before allergen challenge--A0, during the early asthmatic reaction (EAR)--A0 during the late asthmatic reaction (LAR)--A2, and 24 hours after administration of the first allergen dose--A3. The concentrations of TF and TFPI were measured by ELISA method. The concentration of TF in asthmatic patients was significantly higher as compared with healthy controls. In patients that developed only EAR, the concentration of TF increased at A1, then decreased at time when LAR should be developed (A2) and it was comparable with beginning values 24 hours after starting the challenge. In patients that developed EAR and LAR, the mean TF concentration increased during EAR (A,) with subsequent decline during LAR (A2). 24 hours after starting the challenge the TF concentration was higher than beginning values (A3). The concentration of TFPI in patients that developed only EAR was significantly higher than the values in healthy controls. It decreased during EAR (A1) and increased 24 hours after starting the challenge (A3). The mean concentration of TFPI in patients that developed EAR and LAR was not significantly higher than the values in healthy controls. It increased during EAR and LAR with the highest values 24 hours after starting the challenge. Coagulation system seems to be activated during allergic inflammation after allergen challenge. Increased levels of TF and TFPI in asthma patients may be connected with chronic bronchial inflammation and remodeling of bronchial wall.

Topics: Adult; Asthma; Bronchial Provocation Tests; Case-Control Studies; Enzyme-Linked Immunosorbent Assay; Female; Humans; Lipoproteins; Male; Thromboplastin

In addition to its central role in hemostasis, thrombin may play a role in inflammation and remodeling. To investigate the contribution of thrombin to allergic airway inflammation in asthma, we used an enzymatic assay to determine thrombin activity in bronchoalveolar lavage fluid obtained from 19 subjects with atopic asthma before (Day 0) and 48 hours after (Day 2) segmental bronchoprovocation with antigen. Thrombin activity increased from 0 (0, 2.9) on Day 1 to 41.1 (0.3, 75.6) U x 10(-3)/ml on Day 2 (p = 0.002) and correlated with total protein levels in lavage fluid on Day 2 (r = 0.885, p < 0.001). After antigen challenge, thrombin activity also showed significant correlations with interleukin-5 (r = 0.66, p = 0.002), transforming growth factor beta1 (r = 0.70, p < 0.001), fibronectin (r = 0.85, p < 0.001) and tissue factor (r = 0.55, p = 0.03) levels in lavage fluid. Furthermore, Day 2, but not Day 0 lavage fluid, induced proliferation of human airway fibroblasts. This mitogenic effect was significantly reduced with hirudin, a specific thrombin inhibitor. Taken together, our findings suggest that allergen-driven airway inflammation in asthma is associated with enhanced potential for fibroblast proliferation that is related, at least in part, to increased thrombin activity. We propose that enhanced thrombin activity provides a potential link between allergic inflammation and initiation of airway remodeling.

Topics: Adult; Allergens; Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Cohort Studies; Enzyme-Linked Immunosorbent Assay; Female; Fibronectins; Humans; Inflammation; Inflammation Mediators; Interleukin-5; Male; Middle Aged; Probability; Prospective Studies; Sensitivity and Specificity; Statistics, Nonparametric; Thrombin; Thromboplastin; Transforming Growth Factor beta

The mechanism of airway remodeling in asthmatic patients is poorly understood. Thrombin is a multifunctional protease that, in addition to its critical role in thrombotic processes, has also been described as inducing cellular and molecular events relevant to tissue remodeling. The present investigation was undertaken to evaluate the activity of thrombin in the sputum of asthmatic patients and its potential role in airway remodeling. The study population comprised 8 healthy subjects and 14 stable patients with bronchial asthma. The concentrations of thrombin, thrombin-antithrombin complex (TAT), and tissue factor were measured in the sputum of all subjects. The concentrations of thrombin (p = 0. 007), TAT (p = 0.01), and tissue factor (p = 0.02) in sputum were significantly higher in asthmatic patients than in healthy controls. The proliferative effects that sputum from asthmatic patients (p = 0. 01) and thrombin (p = 0.03) have on cultured human smooth muscle cells was inhibited significantly in the presence of recombinant hirudin, a specific thrombin inhibitor. Significant statistical correlation was observed between the degree of bronchial responsiveness and the sputum concentrations of thrombin (r = -0.8; p = 0.02) and TAT (r = -0.9; p = 0.01). The results of this study showed that increased thrombin generation occurs in the airway of patients with asthma and that it may play a role in the pathogenesis of airway remodeling. Further studies should be carried out to assess whether these findings are also observed in other airway diseases.

Topics: Antithrombin III; Asthma; Bronchi; Cells, Cultured; Female; Humans; Male; Middle Aged; Muscle, Smooth; Peptide Hydrolases; Sputum; Thrombin; Thromboplastin