interleukin-8 and Asthma

This meta-analysis aimed to compare the efficacy of montelukast (MKST) combined with budesonide (BUD) and BUD alone in the treatment of pulmonary inflammation and pulmonary function in children with cough variant asthma (CVA). Five electronic databases were searched for studies about MKST+BUD therapy and BUD alone therapy on inflammation and pulmonary function in CVA children from inception to November 23, 2021. Twenty-two articles were included. The results showed that, compared with BUD alone, the combination treatment could achieve better improvement of pulmonary function and lower levels of inflammation (MKST+BUD group: FEV1: SMD = 2.77, 95% CI: 2.07, 3.46; FVC: SMD = 2.54, 95% CI: 1.82, 3.27; PEF: SMD = 2.27, 95% CI: 1.79, 2.75; IgE: SMD = -7.95, 95% CI: -9.66, -6.25; TNF-α: SMD = -4.67, 95% CI: -6.04, -3.31; IL-8: SMD = -8.18, 95% CI: -11.46, -4.90; BUD alone group: FEV1: SMD = 1.83, 95% CI: 1.34, 2.31; FVC: SMD = 1.39, 95% CI: 0.93, 1.84; PEF: SMD = 1.51, 95% CI: 1.13, 1.89; IgE: SMD = -4.93, 95% CI: -6.14, -3.72; TNF-α: SMD = -2.78, 95% CI: -3.76, -1.80; IL-8: SMD = -4.94, 95% CI: -7.10, -2.79). To conclude, compared with BUD alone, MKST+BUD therapy was found to be more effective in improving pulmonary function and reducing inflammation in CVA children. Key Words: Montelukast, Budesonide, Cough variant asthma, Children, Pulmonary function, Inflammatory markers, Meta-analysis.

Topics: Asthma; Budesonide; Child; Cough; Humans; Immunoglobulin E; Inflammation; Interleukin-8; Tumor Necrosis Factor-alpha

Asthma is a complex, heterogeneous condition that affects over 350 million people globally. It is characterised by bronchial hyperreactivity and airways inflammation. A subset display marked airway neutrophilia, associated with worse lung function, higher morbidity and poor response to treatment. In these individuals, recent metagenomic studies have identified persistent bacterial infection, particularly with non-encapsulated strains of the Gram-negative bacterium

Topics: Asthma; Cell Adhesion Molecule-1; Cytokines; Haemophilus influenzae; Humans; Inflammasomes; Interleukin-12; Interleukin-17; Interleukin-6; Interleukin-8; Macrolides; NLR Family, Pyrin Domain-Containing 3 Protein; Pulmonary Disease, Chronic Obstructive; Respiratory System; Tumor Necrosis Factors

Topics: Asthma; Humans; Inflammation; Interleukin-8; Pulmonary Disease, Chronic Obstructive; Quality of Life

Asthma and chronic obstructive pulmonary disease (COPD) cause significant morbidity and mortality worldwide. In the context of disease pathogenesis, both asthma and COPD involve chronic inflammation of the lung and are characterised by the abnormal release of inflammatory cytokines, dysregulated immune cell activity and remodelling of the airways. To date, current treatments still only manage symptoms and do not reverse the primary disease processes. In recent work, interleukin (IL)-1α and IL-1β have been suggested to play important roles in both asthma and COPD. In this review, we summarise overwhelming pre-clinical evidence for dysregulated signalling of IL-1α and IL-1β contributing to disease pathogenesis and discuss the paradox of IL-1 therapeutic studies in asthma and COPD. This is particularly important given recent completed and ongoing clinical trials with IL-1 biologics that have had varying degrees of failure and success as therapeutics for disease modification in asthma and COPD.

Topics: Asthma; Humans; Interleukin-8; Lung; Pulmonary Disease, Chronic Obstructive

Middle East Respiratory Syndrome (MERS) is a novel respiratory illness firstly reported in Saudi Arabia in 2012. It is caused by a new corona virus, called MERS corona virus (MERS-CoV). Most people who have MERS-CoV infection developed severe acute respiratory illness.. This work is done to determine the clinical characteristics and the outcome of intensive care unit (ICU) admitted patients with confirmed MERS-CoV infection.. This study included 32 laboratory confirmed MERS corona virus infected patients who were admitted into ICU. It included 20 (62.50%) males and 12 (37.50%) females. The mean age was 43.99 ± 13.03 years. Diagnosis was done by real-time reverse transcription polymerase chain reaction (rRT-PCR) test for corona virus on throat swab, sputum, tracheal aspirate, or bronchoalveolar lavage specimens. Clinical characteristics, co-morbidities and outcome were reported for all subjects.. Most MERS corona patients present with fever, cough, dyspnea, sore throat, runny nose and sputum. The presence of abdominal symptoms may indicate bad prognosis. Prolonged duration of symptoms before patients' hospitalization, prolonged duration of mechanical ventilation and hospital stay, bilateral radiological pulmonary infiltrates, and hypoxemic respiratory failure were found to be strong predictors of mortality in such patients. Also, old age, current smoking, smoking severity, presence of associated co-morbidities like obesity, diabetes mellitus, chronic heart diseases, COPD, malignancy, renal failure, renal transplantation and liver cirrhosis are associated with a poor outcome of ICU admitted MERS corona virus infected patients.. Plasma HO-1, ferritin, p21, and NQO1 were all elevated at baseline in CKD participants. Plasma HO-1 and urine NQO1 levels each inversely correlated with eGFR (. SnPP can be safely administered and, after its injection, the resulting changes in plasma HO-1, NQO1, ferritin, and p21 concentrations can provide information as to antioxidant gene responsiveness/reserves in subjects with and without kidney disease.. A Study with RBT-1, in Healthy Volunteers and Subjects with Stage 3-4 Chronic Kidney Disease, NCT0363002 and NCT03893799.. HFNC did not significantly modify work of breathing in healthy subjects. However, a significant reduction in the minute volume was achieved, capillary [Formula: see text] remaining constant, which suggests a reduction in dead-space ventilation with flows > 20 L/min. (ClinicalTrials.gov registration NCT02495675).. 3 组患者手术时间、术中显性失血量及术后 1 周血红蛋白下降量比较差异均无统计学意义（. 对于肥胖和超重的膝关节单间室骨关节炎患者，采用 UKA 术后可获满意短中期疗效，远期疗效尚需进一步随访观察。.. Decreased muscle strength was identified at both time points in patients with hEDS/HSD. The evolution of most muscle strength parameters over time did not significantly differ between groups. Future studies should focus on the effectiveness of different types of muscle training strategies in hEDS/HSD patients.. These findings support previous adverse findings of e-cigarette exposure on neurodevelopment in a mouse model and provide substantial evidence of persistent adverse behavioral and neuroimmunological consequences to adult offspring following maternal e-cigarette exposure during pregnancy. https://doi.org/10.1289/EHP6067.. This RCT directly compares a neoadjuvant chemotherapy regimen with a standard CROSS regimen in terms of overall survival for patients with locally advanced ESCC. The results of this RCT will provide an answer for the controversy regarding the survival benefits between the two treatment strategies.. NCT04138212, date of registration: October 24, 2019.. Results of current investigation indicated that milk type and post fermentation cooling patterns had a pronounced effect on antioxidant characteristics, fatty acid profile, lipid oxidation and textural characteristics of yoghurt. Buffalo milk based yoghurt had more fat, protein, higher antioxidant capacity and vitamin content. Antioxidant and sensory characteristics of T. If milk is exposed to excessive amounts of light, Vitamins B. The two concentration of ZnO nanoparticles in the ambient air produced two different outcomes. The lower concentration resulted in significant increases in Zn content of the liver while the higher concentration significantly increased Zn in the lungs (p < 0.05). Additionally, at the lower concentration, Zn content was found to be lower in brain tissue (p < 0.05). Using TEM/EDX we detected ZnO nanoparticles inside the cells in the lungs, kidney and liver. Inhaling ZnO NP at the higher concentration increased the levels of mRNA of the following genes in the lungs: Mt2 (2.56 fold), Slc30a1 (1.52 fold) and Slc30a5 (2.34 fold). At the lower ZnO nanoparticle concentration, only Slc30a7 mRNA levels in the lungs were up (1.74 fold). Thus the two air concentrations of ZnO nanoparticles produced distinct effects on the expression of the Zn-homeostasis related genes.. Until adverse health effects of ZnO nanoparticles deposited in organs such as lungs are further investigated and/or ruled out, the exposure to ZnO nanoparticles in aerosols should be avoided or minimised.

Topics: A549 Cells; Acetylmuramyl-Alanyl-Isoglutamine; Acinetobacter baumannii; Acute Lung Injury; Adaptor Proteins, Signal Transducing; Adenine; Adenocarcinoma; Adipogenesis; Administration, Cutaneous; Administration, Ophthalmic; Adolescent; Adsorption; Adult; Aeromonas hydrophila; Aerosols; Aged; Aged, 80 and over; Aging; Agriculture; Air Pollutants; Air Pollution; Airway Remodeling; Alanine Transaminase; Albuminuria; Aldehyde Dehydrogenase 1 Family; Algorithms; AlkB Homolog 2, Alpha-Ketoglutarate-Dependent Dioxygenase; Alzheimer Disease; Amino Acid Sequence; Ammonia; Ammonium Compounds; Anaerobiosis; Anesthetics, Dissociative; Anesthetics, Inhalation; Animals; Anti-Bacterial Agents; Anti-HIV Agents; Anti-Infective Agents; Anti-Inflammatory Agents; Antibiotics, Antineoplastic; Antibodies, Antineutrophil Cytoplasmic; Antibodies, Monoclonal, Humanized; Antifungal Agents; Antigens, Bacterial; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Antimetabolites, Antineoplastic; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Antioxidants; Antitubercular Agents; Antiviral Agents; Apolipoproteins E; Apoptosis; Arabidopsis; Arabidopsis Proteins; Arsenic; Arthritis, Rheumatoid; Asthma; Atherosclerosis; ATP-Dependent Proteases; Attitude of Health Personnel; Australia; Austria; Autophagy; Axitinib; Bacteria; Bacterial Outer Membrane Proteins; Bacterial Proteins; Bacterial Toxins; Bacterial Typing Techniques; Bariatric Surgery; Base Composition; Bayes Theorem; Benzoxazoles; Benzylamines; beta Catenin; Betacoronavirus; Betula; Binding Sites; Biological Availability; Biological Oxygen Demand Analysis; Biomarkers; Biomarkers, Tumor; Biopsy; Bioreactors; Biosensing Techniques; Birth Weight; Blindness; Blood Chemical Analysis; Blood Gas Analysis; Blood Glucose; Blood Pressure; Blood Pressure Monitoring, Ambulatory; Blood-Brain Barrier; Blotting, Western; Body Mass Index; Body Weight; Bone and Bones; Bone Density; Bone Resorption; Borates; Brain; Brain Infarction; Brain Injuries, Traumatic; Brain Neoplasms; Breakfast; Breast Milk Expression; Breast Neoplasms; Bronchi; Bronchoalveolar Lavage Fluid; Buffaloes; Cadherins; Calcification, Physiologic; Calcium Compounds; Calcium, Dietary; Cannula; Caprolactam; Carbon; Carbon Dioxide; Carboplatin; Carcinogenesis; Carcinoma, Ductal; Carcinoma, Ehrlich Tumor; Carcinoma, Hepatocellular; Carcinoma, Non-Small-Cell Lung; Carcinoma, Pancreatic Ductal; Carcinoma, Renal Cell; Cardiovascular Diseases; Carps; Carrageenan; Case-Control Studies; Catalysis; Catalytic Domain; Cattle; CD8-Positive T-Lymphocytes; Cell Adhesion; Cell Cycle Proteins; Cell Death; Cell Differentiation; Cell Line; Cell Line, Tumor; Cell Movement; Cell Nucleus; Cell Phone Use; Cell Proliferation; Cell Survival; Cell Transformation, Neoplastic; Cell Transformation, Viral; Cells, Cultured; Cellulose; Chemical Phenomena; Chemoradiotherapy; Child; Child Development; Child, Preschool; China; Chitosan; Chlorocebus aethiops; Cholecalciferol; Chromatography, Liquid; Circadian Clocks; 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Forced Expiratory Volume; Forests; Fractures, Bone; Fruit and Vegetable Juices; Fusobacteria; G1 Phase Cell Cycle Checkpoints; G2 Phase Cell Cycle Checkpoints; Gamma Rays; Gastrectomy; Gastrointestinal Microbiome; Gastrointestinal Stromal Tumors; Gefitinib; Gels; Gemcitabine; Gene Amplification; Gene Expression; Gene Expression Regulation; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Neoplastic; Gene Expression Regulation, Plant; Gene Knockdown Techniques; Gene-Environment Interaction; Genotype; Germany; Glioma; Glomerular Filtration Rate; Glucagon; Glucocorticoids; Glycemic Control; Glycerol; Glycogen Synthase Kinase 3 beta; Glycolipids; Glycolysis; Goblet Cells; Gram-Negative Bacterial Infections; Granulocyte Colony-Stimulating Factor; Graphite; Greenhouse Effect; Guanidines; Haemophilus influenzae; HCT116 Cells; Health Knowledge, Attitudes, Practice; Health Personnel; Health Services Accessibility; Health Services Needs and Demand; Health Status Disparities; Healthy Volunteers; 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YAP-Signaling Proteins; Yogurt; Young Adult; Zebrafish; Zebrafish Proteins; Ziziphus

To discuss the presence and role of neutrophils in asthma and allergic diseases, and outline the importance of pollen and cat dander-induced innate neutrophil recruitment in induction of allergic sensitization and allergic inflammation.. Uncontrolled asthma is associated with elevated numbers of neutrophils, and levels of neutrophil-attracting chemokine IL-8 and IL-17 in bronchoalveolar lavage fluids. These parameters negatively correlate with lung function. Pollen allergens and cat dander recruit neutrophils to the airways in a toll-like receptor 4, myeloid differentiation protein-2, and chemokine (C-X-C motif) receptor (CXCR) 2-dependent manner. Repeated recruitment of activated neutrophils by these allergens facilitates allergic sensitization and airway inflammation. Inhibition of neutrophil recruitment with CXCR2 inhibitor, disruption of toll-like receptor 4, or small interfering RNA against myeloid differentiation protein-2 also inhibits allergic inflammation. The molecular mechanisms by which innately recruited neutrophils contribute to shifting the airway inflammatory response induced by allergens from neutrophilic to an eosinophilic-allergic is an area of active research.. Recent studies have revealed that neutrophil recruitment is important in the development of allergic sensitization and inflammation. Inhibition of neutrophils recruitment may be a strategy to control allergic inflammation.

Topics: Allergens; Animals; Asthma; Cats; Humans; Immunity, Innate; Interleukin-17; Interleukin-8; Lymphocyte Antigen 96; Neutrophil Infiltration; Neutrophils; Receptors, Interleukin-8B; Toll-Like Receptor 4

The incidences of childhood allergies have been increasing in recent years in many parts of the world. The development of allergic disorders is attributed to a complex series of interactions between individuals' genetic backgrounds and their immune and psychoneurotic responses to environmental factors. Among the various possible environmental causes of childhood allergies, the early exposure of developing infants to air pollutants and the presence of persistent chemical pollutants such as pesticides have been suggested most frequently. Therefore, it is very important to obtain epidemiological evidence of direct associations between clearly defined adverse health effects and exposure to low levels of pollutants. However, there are no useful biomarkers for assessing such associations. Thus, we planned to establish reliable health-related biomarkers that could be used to investigate these relationships in children. The serum concentrations of several sub-types of polychlorinated biphenyl (PCB) congeners were found to be significantly correlated with interleukin (IL)-8 mRNA expression among asthmatic children. In addition, IL-22 mRNA expression was found to be particularly useful for detecting the effects of environmental pollutants, especially PCB congeners, in a sub-population of vulnerable children who exhibited positive immunoglobulin E (IgE) responses to milk or egg. Furthermore, we detected significant differences in IL-22 mRNA expression between the IgE-negative non-asthmatic subjects and the asthmatic children who exhibited positive IgE reactions toward egg or milk. In conclusion, IL-8 and IL-22 mRNA expressions could be useful biomarkers for detecting sub-populations of children who are particularly vulnerable to the adverse health effects of environmental pollutants, especially PCBs.

Topics: Animals; Asthma; Biomarkers; Child, Preschool; Environmental Exposure; Humans; Hypersensitivity; Immunoglobulin E; Infant; Interleukin-22; Interleukin-8; Interleukins; Japan; Milk; Molecular Epidemiology; Polychlorinated Biphenyls; Real-Time Polymerase Chain Reaction; RNA, Messenger

Neutrophils may play an important role in the pathogenesis of severe asthma. Their infiltration into the airway is increased. Interleukin (IL)-8 is involved in this process, and is actually upregulated in the airways of patients. We have observed that in the absence of eosinophil chemoattractants, neutrophils stimulated by IL-8 augment eosinophil trans-basement membrane migration by releasing superoxide anion, matrix metalloproteinase, leukotriene B(4) and platelet-activating factor. These findings suggest that IL-8-stimulated neutrophils could lead eosinophils to accumulate in the airways of asthmatic patients, which might be a mechanism for corticosteroid resistance in severe asthma. However, the mechanisms of IL-8 upregulation in the airway are not completely understood. Several studies suggest that IL-17 (or T helper 17 cells; Th17) is involved in the IL-8 upregulation observed in severe asthma. We clarified that dopamine induces Th17 differentiation through dopamine D1-like receptor (D1-like-R), and that the D1-like-R antagonist attenuates Th17-mediated diseases like experimental autoimmune encephalomyelitis. Furthermore, we demonstrated that a D1-like-R antagonist significantly suppressed ovalbumin (OVA)-induced neutrophilic airway inflammation in OVA T cell receptor-transgenic DO11.10 mice through inhibiting Th17-mediated immune responses. Therefore, dopamine D1-like-R antagonists could become useful for treating Th17-mediated neutrophil-dominant severe asthma. As inhaled corticosteroids are known to be less effective for controlling neutrophilic inflammation, a more effective therapeutic strategy for neutrophil-dominant asthma should still be elucidated.

Topics: Animals; Asthma; Cell Movement; Humans; Interleukin-8; Neutrophils; Pneumonia; Receptors, Dopamine D1; Th17 Cells

The bronchoalveolar epithelium is submitted to numerous mechanical strains. These strains induce a specific cellular activity at the tissue level. This type of activation has been studied in respiratory medicine, mainly in the context of mechanical ventilation and asthma. The phenomenon of mechanotransduction is linked to various epithelial cellular activities such as epithelium repair, extracellular matrix remodelling, inflammatory mediator release and mucociliary regulation. In this review, the main studies related to bronchoalveolar epithelial mechanotransduction are reported to bring a new perspective on this little known biological phenomenon. A better understanding of the physiological and pathological aspects will potentially offer new treatment approaches for bronchial diseases.

Topics: Apoptosis; Asthma; Bronchi; Cilia; Collagen; Cytokines; Epithelial Cells; Epithelium; ErbB Receptors; Extracellular Matrix; Gene Expression Profiling; Inflammation; Inflammation Mediators; Interleukin-8; Mechanotransduction, Cellular; Mucus; Pulmonary Alveoli; Reactive Oxygen Species; Respiration, Artificial

Inhaled air is contaminated with pathogens and particulates that may deposit in the airways and damage the host. In response to these invaders, the airway epithelium has developed innate immune responses that provide a defence against the invaders and protect the airway structure and function. Thus, the epithelium of conducting airways becomes the "battleground" between the invaders and the host. Recent evidence suggests that airway epithelial surface signalling through the epidermal growth factor receptor (EGFR) is a convergent pathway producing innate immune responses to a variety of infectious and noninfectious noxious stimuli. In the present review, the EGFR signalling pathways leading to airway mucin production, neutrophil recruitment (via interleukin-8 production) and airway epithelial repair were examined. The importance of these findings in human airway diseases was also investigated. The current authors suggest that the exaggerated innate immune responses found in chronic inflammatory airway diseases (e.g. chronic obstructive pulmonary disease, cystic fibrosis and severe asthma) contribute to the pathogenesis or the aggravation of these diseases. Potential therapies include inhibition of the various elements of the described epidermal growth factor receptor cascade. In considering each therapeutic intervention, the potential benefits must be considered in relation to potential deleterious effects.

Topics: Animals; Asthma; Cystic Fibrosis; ErbB Receptors; Humans; Immunity, Innate; Interleukin-8; Ligands; Models, Biological; Mucins; Neutrophils; Pulmonary Disease, Chronic Obstructive; Reactive Oxygen Species; Signal Transduction

Prostaglandin D(2) (PGD(2)), a major prostanoid produced by activated mast cells, has long been implicated in allergic diseases. Recent studies have shown that PGD(2) exerts its effects through two different G-protein-coupled receptors (GPCRs), the D-prostanoid receptor (DP) and the chemoattractant receptor-homologous molecule expressed on T helper type-2 cells (CRTH2), expressed in various human tissues. The PGD(2)/CRTH2 system mediates the chemotaxis of eosinophils, basophils, and Th2 cells, which are involved in the induction of allergic inflammation. We have reported that normal human bronchial epithelial cells (NHBE) and epithelial cell lines (NCI-H(292)) expressed CRTH2, and PGD(2) induces production of IL-8 and GM-CSF. This review discusses the role of CRTH2/DP on epithelial cells and mentions a possible novel receptor for PGD(2).

Topics: Asthma; Bronchi; Bronchitis; Cell Line; Chemotaxis; Epithelial Cells; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; MAP Kinase Signaling System; Organ Specificity; Prostaglandin D2; Receptors, Immunologic; Receptors, Prostaglandin; Respiratory Hypersensitivity; RNA, Messenger; Th2 Cells

Morbidity and mortality of chronic obstructive pulmonary disease (COPD) are considerable and still increasing. The disease is gaining increasing socioeconomic importance. The knowledge of underlying mechanisms is of special relevance because of the lack of a curative therapy. Respiratory infections have been identified as the most important triggers of acute exacerbations but recent data suggest that they might also play an important role in COPD pathogenesis. This knowledge might offer new therapeutic perspectives in the future. The aim of this review is, therefore, to describe the inflammatory processes involved and to specify the role of respiratory infections in this context.

Topics: Asthma; Bacterial Infections; Bronchitis; Common Cold; Disease Progression; Humans; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; Neutrophils; Picornaviridae Infections; Pulmonary Disease, Chronic Obstructive; Respiratory Tract Infections; Rhinovirus; Tumor Necrosis Factor-alpha

Asthma is a heterogeneous disease with multiple phenotypes. There are no tissue or bronchoalveolar lavage biomarkers that are "specific" for asthma. Markers associated with eosinophilic, neutrophilic, and paucigranulocytic asthma are discussed here, and those for remodeling. Efforts are to compare tissue and lavage biomarkers with less invasive measures, such as sputum, serum, or exhaled breath, to improve the treatment and management of asthma.

Topics: Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Eosinophilia; Humans; Interleukin-5; Interleukin-8; Matrix Metalloproteinase 9

Arachidonic acid metabolism represents an important source of mediators with ambivalent actions. Among these, lipoxins (LXs) are the first agents identified and recognized as anti-inflammatory endogenous lipid mediators, which are involved in the resolution of inflammation and are present in the airways of asthmatic patients. Lipoxins result mainly from the interaction between 5 and 15-lipoxygenases (LO) and their levels are modulated by the degree of bronchial inflammation as well as by the long-term glucocorticoid treatments. In the airways, LX synthesis is higher in mild asthmatics than in severe asthmatics, whereas in vitro chemokine release inhibition by LXs is more effective in cells from severe asthmatics than from mild asthmatics. LipoxinA(4) effects on interleukin (IL)-8 released by blood mononuclear cells and on calcium influx in epithelial cells are mediated by the specific receptor ALX. Lipoxin generation by lung epithelial cells depends mainly on 15-LO activity. Mild asthmatics present higher 15-LOb expression at the epithelium level than severe patients, whereas the LX deficit in severe asthma is associated with an up-regulation of the 15-LOa expressions. Therefore, bronchial epithelial cells become a target for therapeutic intervention and LXs represent a potential therapeutic solution for bronchial inflammation resolution in asthma.

Topics: Arachidonic Acid; Asthma; Bronchi; Calcium; Humans; Inflammation Mediators; Interleukin-8; Lipoxins; Respiratory Mucosa; Signal Transduction

Topics: Antigens, Bacterial; Asthma; Epithelial Cells; Humans; In Vitro Techniques; Interleukin-8; Pneumonia, Mycoplasma

Asthma is characterised by a chronic inflammatory process involving the airway wall leading to airflow limitation and bronchial hyperresponsiveness. This review describes the cellular molecular events underlying airway inflammation and remodelling in asthma and focuses on the distinct mechanisms of action of corticosteroids in beta agonists in asthma pathophysiology. In particular it discusses the evidence from studies both in vitro and in vivo to suggest that there is a complimentary beneficial interaction of these agents on the asthma process.

Topics: Administration, Inhalation; Adrenergic Agonists; Adrenergic beta-2 Receptor Agonists; Animals; Anti-Asthmatic Agents; Asthma; Chemokine CCL11; Chemokines, CC; Drug Synergism; Drug Therapy, Combination; Glucocorticoids; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Muscle Relaxation; Myocytes, Smooth Muscle

Neutrophils have been implicated in the pathogenesis of many inflammatory lung diseases, including the acute respiratory distress syndrome, chronic obstructive pulmonary disease and asthma. The CXC chemokine interleukin (IL)-8, is a potent neutrophil recruiting and activating factor and the detection of IL-8 in clinical samples from patients with these diseases has led clinicians to believe that antagonism of IL-8 may be a practicable therapeutic strategy for disease management. Work over the last decade has concentrated on both the molecular mechanisms by which IL-8 is produced in the inflammatory setting and also on the manner in which IL-8 activates the neutrophil. Expression of the IL-8 gene appears to be controlled by several components of the inflammatory milieu. Whilst lipopolysaccharide, IL-1beta and tumor necrosis factor-alpha are capable of augmenting IL-8 production, IL-10 is a potent inhibitor of IL-8 synthesis and appears to play an auto-regulatory role. Regulation of the IL-8 gene is under the control of nuclear factor kappaB which appears to be a primary target for corticosteroid-mediated repression of IL-8 production. IL-8 exerts is effects on neutrophils by binding with high affinity to two receptors on its cell surface, the chemokine receptors CXCR1 and CXCR2. These closely related receptors belong to the superfamily of G-protein coupled receptors, proteins that historically have proved amenable to antagonism by small molecules. The recent descriptions in the literature of highly potent small molecule antagonists of CXCR2 and their success in blocking in vivo trafficking of neutrophils suggest that antagonism of IL-8 at the receptor level is a viable therapeutic strategy. Clinical trials of such compounds will ultimately provide crucial information currently lacking and will define whether or not IL-8 blockade provides future therapy in pulmonary disease.

Topics: Asthma; Humans; Interleukin-8; Pulmonary Disease, Chronic Obstructive; Receptors, Interleukin-8A; Receptors, Interleukin-8B; Respiratory Distress Syndrome

Airway inflammation with eosinophils is now reported to occur not only in asthma but in other airway diseases such as cough variant asthma, chronic cough, atopic cough, episodic symptoms without asthma, allergic rhinitis, and COPD. Although the prevalence of eosinophilic bronchitis (EB) is less than in asthma, the causes, mechanisms and treatment of EB in these conditions appears to be similar to asthma where allergen induced IL-5 secretion and symptoms are readily responsive to inhaled corticosteroids. The prognosis of EB without asthma is not known but it may be a precursor for asthma and, if so, recognition of this syndrome may permit effective treatment and reduction in the rising prevalence of asthma. Induced sputum analysis allows recognition of EB in clinical practice. The place of the asthma treatment paradigm with early and sustained corticosteroid treatment needs to be defined in EB without asthma. Airway wall remodelling can occur in rhinitis, COPD, and cough variant asthma with EB. The mechanisms and long term implications of this complication in EB without asthma need to be clarified.

Topics: Asthma; Bronchitis; Chronic Disease; Cough; Eosinophilia; Humans; Interleukin-5; Interleukin-8; Pulmonary Disease, Chronic Obstructive; Rhinitis, Allergic, Perennial

There is increasing evidence that inflammatory mechanisms other than eosinophilic inflammation may be involved in producing the final common pathway of enhanced bronchial reactivity and reversible airflow obstruction that characterises asthma. A review of the literature has shown that, at most, only 50% of asthma cases are attributable to eosinophilic airway inflammation. It is hypothesised that a major proportion of asthma is based on neutrophilic airway inflammation, possibly triggered by environmental exposure to bacterial endotoxin, particulate air pollution, and ozone, as well as viral infections. If there are indeed two (or more) subtypes of asthma, and if non-eosinophilic (neutrophil mediated) asthma is relatively common, this would have major consequences for the treatment and prevention of asthma since most treatment and prevention strategies are now almost entirely focused on allergic/eosinophilic asthma and allergen avoidance measures, respectively. It is therefore important to study the aetiology of asthma further, including the underlying inflammatory profiles.

Topics: Asthma; Bronchial Hyperreactivity; Environmental Exposure; Eosinophils; Humans; Inflammation; Interleukin-5; Interleukin-8; Occupational Diseases

Recent clinical evidence shows that acute, severe exacerbations of asthma are associated with recruitment and activation of neutrophils in the airways. There is also experimental evidence from rodents that T-lymphocytes are involved in the recruitment of neutrophils following allergen challenge in sensitised airways. This review addresses the potential role of neutrophils and the cytokine interleukin-17 (IL-17) in severe asthma. IL-17 is produced and released as a free protein from T-lymphocytes of the memory (CD45RO+) subset. Evidence from rats in vivo suggests that IL-17 can recruit and activate neutrophils in the airways; the recruitment is mediated by the rat correlate to the neutrophil chemoattractant interleukin-8 (IL-8) macrophage inflammatory protein-2 (MIP-2). Endogenous peptidases modulate neutrophil recruitment by acting on NK-1 receptors in rat airways in vivo. Human bronchial epithelial cells in vitro respond to stimulation with IL-17 by increasing the production and release of the human neutrophil chemoattractant IL-8. This release of IL-8 is functionally significant; it causes neutrophil chemotaxis in vitro. Furthermore, this IL-8 release is sensitive to a glucocorticoid and is potentiated by the pro-inflammatory cytokine, tumour necrosis factor-alpha (TNF-alpha) in vitro. In addition, IL-17 stimulates human bronchial epithelial cells in vitro to release the neutrophil-activating factor IL-6. This effect of IL-17 on IL-6, and IL-8 is in part mediated via mitogen-activated protein kinases. In conclusion, as indicated in rat airways in vivo and in human bronchial epithelial cells in vitro, IL-17 may constitute a link between the activation of certain T-lymphocytes and mobilisation of neutrophils in the airways, via induced release of C-X-C chemokines and tachykinins. Further studies are required to answer the question whether free, soluble IL-17 protein plays this role in the airways of patients with severe asthma.

Topics: Animals; Asthma; Humans; Interleukin-17; Interleukin-8; Leukocyte Common Antigens; Neutrophil Activation; Neutrophils; Rats; T-Lymphocyte Subsets; Tumor Necrosis Factor-alpha

In order to evaluate the role of neutrophils in the pathogenesis of occupational asthma (OA), 15 toluene diisocyanate (TDI)-asthma and six grain dust-asthma patients were recruited. Controls were the same number of subjects showing negative bronchoprovocation test (BPT) and six house dust mite-sensitive asthma. Bronchoscopic biopsy specimens were stained with monoclonal antibodies to mast cell (AA1), eosinophil (EG2), pan T cell (CD3) and neutrophil (NE). Serum neutrophil chemotactic activity (NCA) was measured before and 10-420 min after BPT. Sputum interleukin-8 (IL-8) and myeloperoxidase (MPO) were also measured. There was a significant increase of NE+ cells as well as AA1+ and EG2+ cells in grain dust- and TDI-asthma compared with house dust-sensitive asthma (P < 0.05). Neutrophil+ cells and AA1+ cells showed a significant correlation in TDI-asthma (r = 0.73, P = 0.02). Serum NCA was significantly increased at 10 min after BPT and decreased at 60 min in subjects with TDI-asthma. In grain dust-asthma, serum NCA increased at 30 min and decreased at 240 min after BPT (P < 0.05). Sputum IL-8 and MPO were significantly increased after BPT in both TDI- and grain dust-asthma (P < 0.05). These findings suggested that neutrophils in the lungs might contribute to bronchoconstriction induced by either TDI or grain dust. The possible involvement of IL-8 in activation of neutrophils was also suggested.

Topics: Asthma; Dust; Edible Grain; Humans; Interleukin-8; Neutrophil Activation; Occupational Diseases

Topics: Asthma; Humans; Interleukin-8; Occupational Diseases

The local inflammatory response that occurs after repeated exposure to allergens or during the late-phase reaction results from a complex network of interactions between inflammatory cells (mast cells, eosinophils, macrophages) and resident cells belonging to the lung structure itself like EC, fibroblasts, or bronchial epithelial cells. Among structural cells, EC represent critical elements: they control leukocyte traffic through the expression of adhesion molecules; they are also able to amplify leukocyte activation through the production of proinflammatory cytokines like IL-1, IL-6, or of chemokines like IL-8. Three cell models have been successively considered. When supernatants of alveolar macrophages, recovered from patients exhibiting a late asthmatic response after allergen exposure, were tested on HUVEC cultures, a TNF alpha-dependent ICAM-1 and E-selectin overexpression was observed. Among mast-cell mediators, histamine was already known to induce a rapid and transient expression of P-selectin; we demonstrated that histamine also induced an IL-6 and IL-8 secretion by HUVEC, which was concentration-dependent and inhibited by H1 or H2 receptor antagonists. Finally purified eosinophils obtained from donors with hypereosinophilia similarly increased adhesion molecule expression and chemokine production. The precise nature of the eosinophil product(s) involved in this process is currently under investigation.

Topics: Asthma; Bronchi; Cell Adhesion Molecules; Cell Communication; Endothelium, Vascular; Eosinophils; Humans; Hypersensitivity; Interleukin-8; Macrophages, Alveolar; Mast Cells; Microcirculation; Models, Biological; T-Lymphocytes, Cytotoxic; Tumor Necrosis Factor-alpha

Rhinoviruses are the cause of the majority of common colds, but their role in lower respiratory disorders is less clear. Recent studies using the polymerase chain reaction to detect rhinoviruses have established respiratory viral infections as major factors in the induction of acute exacerbations of asthma in both adults and children, both in mild exacerbations and in more severe exacerbations leading to hospital admission. Rhinoviruses were the major virus type detected in these studies, accounting for two-thirds of viruses detected. It is not known whether rhinoviruses produce their effects by directly invading the lower airway or by indirect means. Previous clinical studies provide some evidence that rhinoviruses are capable of infecting the lower airway. However, the immunologic response, both in the upper and lower airways, remains poorly defined. Recent studies have provided evidence of increased cellular activation in peripheral blood and in bronchial biopsies in atopic subjects compared with normal subjects during experimental rhinovirus infections. The reasons for these different cellular responses are unclear. Rhinoviruses as well as other respiratory viruses have been shown to increase levels of a variety of cytokines from respiratory epithelium, monocytes, or macrophages. Prominent among these cytokines is interleukin (IL)-8. We have detected increased levels of IL-8 in nasal secretions from subjects with wild-type rhinovirus infections. We studied the mechanisms of rhinovirus-induced IL-8 production and and found protein release from both pulmonary epithelial and peripheral blood mononuclear cells. This protein production was accompanied by increased mRNA expression and evidence of infection of both pulmonary epithelial and monocyte cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Asthma; Case-Control Studies; Child; Cytokines; Hospitalization; Humans; Interleukin-8; Picornaviridae Infections; Polymerase Chain Reaction; Rhinovirus; Seasons

Topics: Animals; Asthma; Blood Platelets; Eosinophils; Humans; Inflammation; Interleukin-8; Lung Diseases

Topics: Asthma; Asthma, Exercise-Induced; Bronchial Provocation Tests; Chemotactic Factors; Histamine Release; Humans; Interleukin-8; Leukocytes; Neutrophils; Physical Exertion

Topics: Airway Obstruction; Asthma; Asthma, Exercise-Induced; Chemotactic Factors; Cold Temperature; Dehydration; Epithelium; Humans; Interleukin-8; Muscle, Smooth; Parasympathetic Nervous System

Topics: Allergens; Asthma; Bronchial Provocation Tests; Chemotactic Factors; Humans; Interleukin-8; Lung; Neutrophils; Physical Exertion

Topics: Asthma; Asthma, Exercise-Induced; Bronchi; Chemotactic Factors; Humans; Hyperventilation; Interleukin-8; Neutrophils; Physical Exertion; Sports; Vagus Nerve

Topics: Allergens; Asthma; Asthma, Exercise-Induced; Chemotactic Factors; Humans; Interleukin-8; Lung; Molecular Weight

Exercise-induced asthma is considered in terms of the stimulus, the intermediary pathway and the response. Various controversies about each of these components are discussed. The stimulus may be cooling of the airways, loss of water or neither of these, and there is evidence for and against the identity of exercise- and hyperventilation-induced asthma. The intermediary pathway seems certain to involve the release of chemical mediators, although other neurogenic mechanisms have been proposed. Since the response is far from uniform, it may well be that different pathways are involved in different subjects. The effector mechanism appears to be bronchospasm, but recent evidence has suggested that an inflammatory response may be involved in a late reaction to exercise. The variability of exercise-induced asthma may well be due to variations in intrinsic bronchial reactivity resulting from allergenic stimulation.

Topics: Asthma; Asthma, Exercise-Induced; Bicycling; Body Temperature Regulation; Bronchial Spasm; Chemotactic Factors; Humans; Humidity; Interleukin-8; Reflex, Abnormal; Respiration; Running; Swimming; Vagus Nerve

Topics: Animals; Asthma; Bronchial Provocation Tests; Bronchial Spasm; Chemotactic Factors; Humans; Inflammation; Interleukin-8; Leukocytes; Receptors, Complement; Receptors, Complement 3b

Topics: Animals; Arachidonic Acids; Asthma; Basophils; Biogenic Amines; Bronchi; Chemotactic Factors; Guinea Pigs; Humans; Inflammation; Interleukin-8; Lipoxygenase; Lung; Mast Cells; Mice; Rabbits; Rats; Receptors, IgE; Receptors, Immunologic; SRS-A

Topics: Anaphylaxis; Animals; Asthma; Chemical Phenomena; Chemistry; Chemotactic Factors; Food Hypersensitivity; Guinea Pigs; Humans; Inflammation; Interleukin-8; Lung; Mast Cells; Time Factors

The events which lead to airway narrowing in bronchial asthma are complex. There is little doubt that mast cell-derived pharmacological agents are involved, at least in part, in the initiation of the asthmatic response. However, the inflammatory response which follows mast cell activation might have more relevance to the daily pattern of asthma than the direct effects of mediators on bronchial tissue. Although the IgE mediated release of mediators from sensitized mast cells seems to play a role in pathogenesis in some individuals for some of the time, there is now increasing awareness that mast cells are also triggered by a number of non-immunological stimuli such as exercise/cold air, infection and agents which activate the complement system. Mast cell mediators are either pre-formed within granules or generated from membrane-bound phospholipids. The pre-formed mediators include histamine, various chemotactic peptides including ECF-A and the high molecular weight neutrophil chemotactic factor (NCF), proteases, glycosidases, and the heparin proteoglycan. The membrane-derived agents include the lipoxygenase products (e.g. LTB4 and the "SRS-A" leukotrienes-LTC4/D4/E4), prostaglandins and thromboxane in addition to the PAF-ace-tether (AGEPC). The mediators are diverse both in chemical composition and modes of actions. However, many of the pathological features of bronchial asthma can be explained on the basis of their recognised actions. These can be summarised as follows. Bronchial smooth muscle constriction (histamine, LTC4, LTD4, LTE4, PGF2 alfa, PGD2 and PAF); mucosal oedema (increased permeability--histamine, LTC4, LTD4 and PAF; vasodilation--PGD2, PGE2); mucous plugging (histamine, mono-HETEs and LTC4); inflammatory cell infiltrate (NCF, ECF-A peptides, HETEs, LTB4 and PAF); desquamation of epithelium (proteases, glycosidases, together with lysosomal enzymes, and basic proteins derived from neutrophils and eosinophils). It is likely that mild, easily reversible, episodic asthma is due largely to bronchial smooth muscle contraction whereas the late sustained response is more indicative of an inflammatory response, and dependent on the infiltration of neutrophils and eosinophils as the result of mediators which recruit and activate leucocytes. Much of the evidence for this is based on the demonstration that NCF concentrations in the serum are elevated during early and late phase, antigen- and exercise-induced asthma. Moderate to severe asthma is likely t

Topics: Animals; Arachidonic Acid; Arachidonic Acids; Asthma; Asthma, Exercise-Induced; Bronchitis; Chemotactic Factors; Humans; Hypersensitivity; Immunoglobulin E; Interleukin-8; Mast Cells; Platelet Activating Factor; SRS-A

Neutrophil chemotactic factor (NCF) is a slight acidic macromolecule which is released into the circulation of asthmatic individuals following antigen provocation and an exercise task. It also appears in late asthmatic reactions with a time course of appearance which precedes the second fall in FEV1. The release of NCF was inhibited by prior administration of disodium cromoglycate (DSCG) suggesting its possible mast cell origin.

Topics: Airway Obstruction; Asthma; Asthma, Exercise-Induced; Basophils; Bronchial Provocation Tests; Chemical Phenomena; Chemistry; Chemotactic Factors; Chronic Disease; Forced Expiratory Volume; Histamine Release; Humans; Inflammation; Interleukin-8

Topics: Administration, Intranasal; Airway Obstruction; Animals; Asthma; Bronchial Spasm; Chemotactic Factors; Clinical Trials as Topic; Cromolyn Sodium; Double-Blind Method; Guinea Pigs; Haplorhini; Histamine Release; Humans; Interleukin-8; Long-Term Care; Mast Cells; Sulfur Dioxide; Theophylline

Abnormalities of the airway smooth muscle (ASM) layer in asthma may develop before birth. We hypothesize that antenatal inflammation causes physiological abnormalities of the ASM that predisposes asthma. This study determined the short-term effects of antenatal inflammation on the developing ASM. Fourteen pregnant ewes were randomly assigned to one of three groups. Fetal lambs were exposed to intra-amniotic injections of lipopolysaccharide (LPS,

Topics: Acetylcholine; Animals; Asthma; Female; Inflammation; Interleukin-8; Lipopolysaccharides; Muscle Contraction; Muscle, Smooth; Pregnancy; Pregnancy Complications; Premature Birth; Sheep

Activation of immunological and systemic inflammation markers are common in obesity and asthma.. The target of this study was to assess impact of weight reduction on immunological and systemic inflammation markers in obese asthma patients.. Eighty asthmatic patients of both sex; their age and body mass index (BMI) mean were 38.72 ± 7.14 year and 32.65 ± 3.18 Kg/m2 respectively. Exclusion criteria included smokers, infections, vaccinations, cancer, surgery, immune system disorders and medications that may influence immune system function as anti-inflammatory medications, analgesics and anti-depressant. All subjects were randomly enrolled in weight reduction group (group A) or control group (group B).. The main findings in the present study indicated that weight reducing program in group (A) was associated with significant reduction in the mean values of IL6, TNF-α, and IL8 in addition to significant increase in the mean values of CD4 and CD8 cell count . However, findings of group (B) showed no significant changes. Moreover, Comparison between both groups at the end of the study revealed significant differences.. Weight reduction improved immunological and systemic inflammation markers in obese asthma patients.

Topics: Adult; Asthma; Biomarkers; Body Mass Index; Cytokines; Diet, Reducing; Exercise; Female; Flow Cytometry; Humans; Immune System; Inflammation; Interleukin-6; Interleukin-8; Male; Middle Aged; Obesity; Systemic Inflammatory Response Syndrome; Treatment Outcome; Weight Loss; Weight Reduction Programs

Allergic asthma is a chronically relapsing inflammatory airway disease with a complex pathophysiology.. This study was undertaken to investigate the potential contribution of NOD2 signaling, proinflammatory cytokines, chitotriosidase (CHIT1) activity, oxidative stress and DNA damage to atopic asthma pathogenesis, as well as to explore their possible role as surrogate noninvasive biomarkers for monitoring asthma severity.. Sixty patients with atopic bronchial asthma who were divided according to asthma severity into 40 mild-moderate, 20 severe atopic asthmatics, in addition to thirty age-matched healthy controls were enrolled in this study. NOD2 expression in PBMCs was assessed by quantitative real-time RT-PCR. DNA damage indices were assessed by alkaline comet assay. Serum IgE, IL-17, IL-8 and 3-Nitrotyrosine levels were estimated by ELISA. Serum CHIT1and GST activities, as well as MDA levels, were measured.. NOD2 mRNA relative expression levels were significantly decreased in atopic asthmatic cases relative to controls with lower values among severe atopic asthmatics. On the other hand, IL-17 and IL-8 serum levels, CHIT1 activity, DNA damage indices and oxidative stress markers were significantly increased in atopic asthmatic cases relative to controls with higher values among severe atopic asthmatics. The change in these parameters correlated significantly with the degree of decline in lung function.. The interplay between NOD2 signaling, proinflammatory cytokines, CHIT1 activity, heightened oxidative stress and DNA damage orchestrates allergic airway inflammation and thus contributing to the pathogenesis of atopic asthma. These parameters qualified for measurement as part of new noninvasive biomarker panels for monitoring asthma severity.

Topics: Adult; Asthma; DNA Damage; Female; Gene Expression Regulation, Enzymologic; Hexosaminidases; Humans; Immunoglobulin E; Inflammation; Interleukin-17; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Nod2 Signaling Adaptor Protein; Oxidation-Reduction; Oxidative Stress; Severity of Illness Index; Tyrosine

The objective of this study was to observe the outcomes of adding an antimicrobial treatment to a conventional treatment regime in horses with severe equine asthma in a clinical setting. Eleven client-owned horses with a history consistent with severe equine asthma, increased respiratory effort and nostril flaring, ≥ 20% neutrophils on bronchoalveolar lavage (BAL), and a positive tracheal wash (TW) bacterial culture were treated with environmental management, corticosteroids, and bronchodilators. Six horses were also treated with an antimicrobial (principal group), while the other 5 were administered saline as a placebo (control group). Treatment with antimicrobials significantly improved the post-treatment clinical score of the principal group compared with the pre-treatment score, whereas no significant difference occurred in the control group. The principal group also had significantly less neutrophil myeloperoxidase (MPO) activity post-treatment than pre-treatment, with a median difference of -0.39 units/[protein] in the principal group and a median difference of -0.21 units/[protein] in the controls. There was no difference in MPO activity pre-. L’objectif de la présente étude était d’observer dans un contexte clinique les résultats de l’ajout d’un traitement antimicrobien au traitement conventionnel de chevaux souffrant d’asthme sévère. Onze chevaux appartenant à des propriétaires et ayant une histoire correspondant avec de l’asthme sévère, un effort inspiratoire augmenté et un élargissement des narines, ≥ 20 % de neutrophiles dans le lavage bronchoalvéolaire (LBA), et une culture bactérienne positive à partir du lavage trachéal (LT) ont été traités par gestion de leur environnement, des corticostéroïdes, et des broncho-dilatateurs. Six chevaux ont également été traités avec un antimicrobien (groupe principal) alors que les cinq autres chevaux ont reçu de la saline à titre de placebo (groupe témoin). Le traitement avec les antimicrobiens améliora de manière significative le score clinique post-traitement du groupe principal comparativement au score pré-traitement, alors qu’aucune différence significative ne fut notée dans le groupe témoin. Dans le groupe principal on nota également qu’il y avait significativement moins d’activité myéloperoxydase (MPO) des neutrophiles post-traitement comparativement à pré-traitement, avec une différence médiane de −0,39 unités/[protéine] dans le groupe principal et une différence médiane de −0,21 unités/[protéine] dans le groupe témoin. Il n’y avait pas de différence de l’activité MPO pré-

Topics: Animals; Asthma; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Female; Horse Diseases; Horses; Interleukin-8; Male; Neutrophils; Peroxidase; Trachea; Tumor Necrosis Factor-alpha

Middle East Respiratory Syndrome (MERS) is a novel respiratory illness firstly reported in Saudi Arabia in 2012. It is caused by a new corona virus, called MERS corona virus (MERS-CoV). Most people who have MERS-CoV infection developed severe acute respiratory illness.. This work is done to determine the clinical characteristics and the outcome of intensive care unit (ICU) admitted patients with confirmed MERS-CoV infection.. This study included 32 laboratory confirmed MERS corona virus infected patients who were admitted into ICU. It included 20 (62.50%) males and 12 (37.50%) females. The mean age was 43.99 ± 13.03 years. Diagnosis was done by real-time reverse transcription polymerase chain reaction (rRT-PCR) test for corona virus on throat swab, sputum, tracheal aspirate, or bronchoalveolar lavage specimens. Clinical characteristics, co-morbidities and outcome were reported for all subjects.. Most MERS corona patients present with fever, cough, dyspnea, sore throat, runny nose and sputum. The presence of abdominal symptoms may indicate bad prognosis. Prolonged duration of symptoms before patients' hospitalization, prolonged duration of mechanical ventilation and hospital stay, bilateral radiological pulmonary infiltrates, and hypoxemic respiratory failure were found to be strong predictors of mortality in such patients. Also, old age, current smoking, smoking severity, presence of associated co-morbidities like obesity, diabetes mellitus, chronic heart diseases, COPD, malignancy, renal failure, renal transplantation and liver cirrhosis are associated with a poor outcome of ICU admitted MERS corona virus infected patients.. Plasma HO-1, ferritin, p21, and NQO1 were all elevated at baseline in CKD participants. Plasma HO-1 and urine NQO1 levels each inversely correlated with eGFR (. SnPP can be safely administered and, after its injection, the resulting changes in plasma HO-1, NQO1, ferritin, and p21 concentrations can provide information as to antioxidant gene responsiveness/reserves in subjects with and without kidney disease.. A Study with RBT-1, in Healthy Volunteers and Subjects with Stage 3-4 Chronic Kidney Disease, NCT0363002 and NCT03893799.. HFNC did not significantly modify work of breathing in healthy subjects. However, a significant reduction in the minute volume was achieved, capillary [Formula: see text] remaining constant, which suggests a reduction in dead-space ventilation with flows > 20 L/min. (ClinicalTrials.gov registration NCT02495675).. 3 组患者手术时间、术中显性失血量及术后 1 周血红蛋白下降量比较差异均无统计学意义（. 对于肥胖和超重的膝关节单间室骨关节炎患者，采用 UKA 术后可获满意短中期疗效，远期疗效尚需进一步随访观察。.. Decreased muscle strength was identified at both time points in patients with hEDS/HSD. The evolution of most muscle strength parameters over time did not significantly differ between groups. Future studies should focus on the effectiveness of different types of muscle training strategies in hEDS/HSD patients.. These findings support previous adverse findings of e-cigarette exposure on neurodevelopment in a mouse model and provide substantial evidence of persistent adverse behavioral and neuroimmunological consequences to adult offspring following maternal e-cigarette exposure during pregnancy. https://doi.org/10.1289/EHP6067.. This RCT directly compares a neoadjuvant chemotherapy regimen with a standard CROSS regimen in terms of overall survival for patients with locally advanced ESCC. The results of this RCT will provide an answer for the controversy regarding the survival benefits between the two treatment strategies.. NCT04138212, date of registration: October 24, 2019.. Results of current investigation indicated that milk type and post fermentation cooling patterns had a pronounced effect on antioxidant characteristics, fatty acid profile, lipid oxidation and textural characteristics of yoghurt. Buffalo milk based yoghurt had more fat, protein, higher antioxidant capacity and vitamin content. Antioxidant and sensory characteristics of T. If milk is exposed to excessive amounts of light, Vitamins B. The two concentration of ZnO nanoparticles in the ambient air produced two different outcomes. The lower concentration resulted in significant increases in Zn content of the liver while the higher concentration significantly increased Zn in the lungs (p < 0.05). Additionally, at the lower concentration, Zn content was found to be lower in brain tissue (p < 0.05). Using TEM/EDX we detected ZnO nanoparticles inside the cells in the lungs, kidney and liver. Inhaling ZnO NP at the higher concentration increased the levels of mRNA of the following genes in the lungs: Mt2 (2.56 fold), Slc30a1 (1.52 fold) and Slc30a5 (2.34 fold). At the lower ZnO nanoparticle concentration, only Slc30a7 mRNA levels in the lungs were up (1.74 fold). Thus the two air concentrations of ZnO nanoparticles produced distinct effects on the expression of the Zn-homeostasis related genes.. Until adverse health effects of ZnO nanoparticles deposited in organs such as lungs are further investigated and/or ruled out, the exposure to ZnO nanoparticles in aerosols should be avoided or minimised.

Topics: A549 Cells; Acetylmuramyl-Alanyl-Isoglutamine; Acinetobacter baumannii; Acute Lung Injury; Adaptor Proteins, Signal Transducing; Adenine; Adenocarcinoma; Adipogenesis; Administration, Cutaneous; Administration, Ophthalmic; Adolescent; Adsorption; Adult; Aeromonas hydrophila; Aerosols; Aged; Aged, 80 and over; Aging; Agriculture; Air Pollutants; Air Pollution; Airway Remodeling; Alanine Transaminase; Albuminuria; Aldehyde Dehydrogenase 1 Family; Algorithms; AlkB Homolog 2, Alpha-Ketoglutarate-Dependent Dioxygenase; Alzheimer Disease; Amino Acid Sequence; Ammonia; Ammonium Compounds; Anaerobiosis; Anesthetics, Dissociative; Anesthetics, Inhalation; Animals; Anti-Bacterial Agents; Anti-HIV Agents; Anti-Infective Agents; Anti-Inflammatory Agents; Antibiotics, Antineoplastic; Antibodies, Antineutrophil Cytoplasmic; Antibodies, Monoclonal, Humanized; Antifungal Agents; Antigens, Bacterial; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Antimetabolites, Antineoplastic; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Antioxidants; Antitubercular Agents; Antiviral Agents; Apolipoproteins E; Apoptosis; Arabidopsis; Arabidopsis Proteins; Arsenic; Arthritis, Rheumatoid; Asthma; Atherosclerosis; ATP-Dependent Proteases; Attitude of Health Personnel; Australia; Austria; Autophagy; Axitinib; Bacteria; Bacterial Outer Membrane Proteins; Bacterial Proteins; Bacterial Toxins; Bacterial Typing Techniques; Bariatric Surgery; Base Composition; Bayes Theorem; Benzoxazoles; Benzylamines; beta Catenin; Betacoronavirus; Betula; Binding Sites; Biological Availability; Biological Oxygen Demand Analysis; Biomarkers; Biomarkers, Tumor; Biopsy; Bioreactors; Biosensing Techniques; Birth Weight; Blindness; Blood Chemical Analysis; Blood Gas Analysis; Blood Glucose; Blood Pressure; Blood Pressure Monitoring, Ambulatory; Blood-Brain Barrier; Blotting, Western; Body Mass Index; Body Weight; Bone and Bones; Bone Density; Bone Resorption; Borates; Brain; Brain Infarction; Brain Injuries, Traumatic; Brain Neoplasms; Breakfast; Breast Milk Expression; Breast Neoplasms; Bronchi; Bronchoalveolar Lavage Fluid; Buffaloes; Cadherins; Calcification, Physiologic; Calcium Compounds; Calcium, Dietary; Cannula; Caprolactam; Carbon; Carbon Dioxide; Carboplatin; Carcinogenesis; Carcinoma, Ductal; Carcinoma, Ehrlich Tumor; Carcinoma, Hepatocellular; Carcinoma, Non-Small-Cell Lung; Carcinoma, Pancreatic Ductal; Carcinoma, Renal Cell; Cardiovascular Diseases; Carps; Carrageenan; Case-Control Studies; Catalysis; Catalytic Domain; Cattle; CD8-Positive T-Lymphocytes; Cell Adhesion; Cell Cycle Proteins; Cell Death; Cell Differentiation; Cell Line; Cell Line, Tumor; Cell Movement; Cell Nucleus; Cell Phone Use; Cell Proliferation; Cell Survival; Cell Transformation, Neoplastic; Cell Transformation, Viral; Cells, Cultured; Cellulose; Chemical Phenomena; Chemoradiotherapy; Child; Child Development; Child, Preschool; China; Chitosan; Chlorocebus aethiops; Cholecalciferol; Chromatography, Liquid; Circadian Clocks; Circadian Rhythm; Circular Dichroism; Cisplatin; Citric Acid; Clinical Competence; Clinical Laboratory Techniques; Clinical Trials, Phase I as Topic; Clinical Trials, Phase II as Topic; Clostridioides difficile; Clostridium Infections; Coculture Techniques; Cohort Studies; Cold Temperature; Colitis; Collagen Type I; Collagen Type I, alpha 1 Chain; Collagen Type XI; Color; Connective Tissue Diseases; Copper; Coronary Angiography; Coronavirus 3C Proteases; Coronavirus Infections; Cost of Illness; Counselors; COVID-19; COVID-19 Testing; Creatine Kinase; Creatinine; Cross-Over Studies; Cross-Sectional Studies; Cryoelectron Microscopy; Cryosurgery; Crystallography, X-Ray; Cues; Cultural Competency; Cultural Diversity; Curriculum; Cyclic AMP Response Element-Binding Protein; Cyclin-Dependent Kinase Inhibitor p21; Cycloparaffins; Cysteine Endopeptidases; Cytokines; Cytoplasm; Cytoprotection; Databases, Factual; Denitrification; Deoxycytidine; Diabetes Complications; Diabetes Mellitus; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diagnosis, Differential; Diatoms; Diet; Diet, High-Fat; Dietary Exposure; Diffusion Magnetic Resonance Imaging; Diketopiperazines; Dipeptidyl Peptidase 4; Dipeptidyl-Peptidase IV Inhibitors; Disease Models, Animal; Disease Progression; Disease-Free Survival; DNA; DNA Damage; DNA Glycosylases; DNA Repair; DNA-Binding Proteins; DNA, Bacterial; DNA, Viral; Docetaxel; Dose Fractionation, Radiation; Dose-Response Relationship, Drug; Down-Regulation; Doxorubicin; Drosophila; Drosophila melanogaster; Drug Carriers; Drug Delivery Systems; Drug Liberation; Drug Repositioning; Drug Resistance, Bacterial; Drug Resistance, Multiple, Bacterial; Drug Resistance, Neoplasm; Drug Screening Assays, Antitumor; Drug Synergism; Drug Therapy, Combination; Edema; Edible Grain; Education, Graduate; Education, Medical, Graduate; Education, Pharmacy; Ehlers-Danlos Syndrome; Electron Transport Complex III; Electron Transport Complex IV; Electronic Nicotine Delivery Systems; Emergency Service, Hospital; Empathy; Emulsions; Endothelial Cells; Endurance Training; Energy Intake; Enterovirus A, Human; Environment; Environmental Monitoring; Enzyme Assays; Enzyme Inhibitors; Epithelial Cells; Epithelial-Mesenchymal Transition; Epoxide Hydrolases; Epoxy Compounds; Erythrocyte Count; Erythrocytes; Escherichia coli; Escherichia coli Infections; Escherichia coli Proteins; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Esophagectomy; Estrogens; Etanercept; Ethiopia; Ethnicity; Ethylenes; Exanthema; Exercise; Exercise Test; Exercise Tolerance; Extracellular Matrix; Extracorporeal Membrane Oxygenation; Eye Infections, Fungal; False Negative Reactions; Fatty Acids; Fecal Microbiota Transplantation; Feces; Female; Femur Neck; Fermentation; Ferritins; Fetal Development; Fibroblast Growth Factor-23; Fibroblast Growth Factors; Fibroblasts; Fibroins; Fish Proteins; Flavanones; Flavonoids; Focus Groups; Follow-Up Studies; Food Handling; Food Supply; Food, Formulated; Forced Expiratory Volume; Forests; Fractures, Bone; Fruit and Vegetable Juices; Fusobacteria; G1 Phase Cell Cycle Checkpoints; G2 Phase Cell Cycle Checkpoints; Gamma Rays; Gastrectomy; Gastrointestinal Microbiome; Gastrointestinal Stromal Tumors; Gefitinib; Gels; Gemcitabine; Gene Amplification; Gene Expression; Gene Expression Regulation; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Neoplastic; Gene Expression Regulation, Plant; Gene Knockdown Techniques; Gene-Environment Interaction; Genotype; Germany; Glioma; Glomerular Filtration Rate; Glucagon; Glucocorticoids; Glycemic Control; Glycerol; Glycogen Synthase Kinase 3 beta; Glycolipids; Glycolysis; Goblet Cells; Gram-Negative Bacterial Infections; Granulocyte Colony-Stimulating Factor; Graphite; Greenhouse Effect; Guanidines; Haemophilus influenzae; HCT116 Cells; Health Knowledge, Attitudes, Practice; Health Personnel; Health Services Accessibility; Health Services Needs and Demand; Health Status Disparities; Healthy Volunteers; 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Infliximab; Infusions, Intravenous; Inhibitory Concentration 50; Injections; Insecticides; Insulin-Like Growth Factor Binding Protein 5; Insulin-Secreting Cells; Interleukin-1; Interleukin-17; Interleukin-8; Internship and Residency; Intestines; Intracellular Signaling Peptides and Proteins; Ion Transport; Iridaceae; Iridoid Glucosides; Islets of Langerhans Transplantation; Isodon; Isoflurane; Isotopes; Italy; Joint Instability; Ketamine; Kidney; Kidney Failure, Chronic; Kidney Function Tests; Kidney Neoplasms; Kinetics; Klebsiella pneumoniae; Knee Joint; Kruppel-Like Factor 4; Kruppel-Like Transcription Factors; Lactate Dehydrogenase 5; Laparoscopy; Laser Therapy; Lasers, Semiconductor; Lasers, Solid-State; Laurates; Lead; Leukocyte L1 Antigen Complex; Leukocytes, Mononuclear; Light; Lipid Peroxidation; Lipopolysaccharides; Liposomes; Liver; Liver Cirrhosis; Liver Neoplasms; Liver Transplantation; Locomotion; Longitudinal Studies; Lopinavir; Lower Urinary Tract Symptoms; Lubricants; Lung; 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Oocytes; Open Reading Frames; Osteoclasts; Osteogenesis; Osteoporosis; Osteoporosis, Postmenopausal; Outpatients; Ovarian Neoplasms; Ovariectomy; Overweight; Oxazines; Oxidants; Oxidation-Reduction; Oxidative Stress; Oxides; Oxidoreductases; Oxygen; Oxygen Inhalation Therapy; Oxygenators, Membrane; Ozone; Paclitaxel; Paenibacillus; Pain Measurement; Palliative Care; Pancreatic Neoplasms; Pandemics; Parasympathetic Nervous System; Particulate Matter; Pasteurization; Patient Preference; Patient Satisfaction; Pediatric Obesity; Permeability; Peroxiredoxins; Peroxynitrous Acid; Pharmaceutical Services; Pharmacists; Pharmacy; Phaseolus; Phenotype; Phoeniceae; Phosphates; Phosphatidylinositol 3-Kinases; Phospholipid Transfer Proteins; Phospholipids; Phosphorus; Phosphorylation; Photoperiod; Photosynthesis; Phylogeny; Physical Endurance; Physicians; Pilot Projects; Piperidines; Pituitary Adenylate Cyclase-Activating Polypeptide; Plant Extracts; Plant Leaves; Plant Proteins; Plant Roots; Plaque, Atherosclerotic; 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Proto-Oncogene Proteins c-ret; Proto-Oncogene Proteins p21(ras); Proton Pumps; Protons; Protoporphyrins; Pseudomonas aeruginosa; Pseudomonas fluorescens; Pulmonary Artery; Pulmonary Disease, Chronic Obstructive; Pulmonary Gas Exchange; Pulmonary Veins; Pyrazoles; Pyridines; Pyrimidines; Qualitative Research; Quinoxalines; Rabbits; Random Allocation; Rats; Rats, Sprague-Dawley; Rats, Wistar; Receptors, Histamine H3; Receptors, Immunologic; Receptors, Transferrin; Recombinant Proteins; Recurrence; Reference Values; Referral and Consultation; Regional Blood Flow; Registries; Regulon; Renal Insufficiency, Chronic; Reperfusion Injury; Repressor Proteins; Reproducibility of Results; Republic of Korea; Research Design; Resistance Training; Respiration, Artificial; Respiratory Distress Syndrome; Respiratory Insufficiency; Resuscitation; Retinal Dehydrogenase; Retreatment; Retrospective Studies; Reverse Transcriptase Inhibitors; Rhinitis, Allergic; Ribosomal Proteins; Ribosomes; Risk Assessment; 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YAP-Signaling Proteins; Yogurt; Young Adult; Zebrafish; Zebrafish Proteins; Ziziphus

The efficacy of budesonide/formoterol maintenance and reliever therapy (BFMRT) in asthma control is well documented in large randomized controlled trials. However, the acute reliever effects and real-life effectiveness are seldom reported.. This multicenter trial enrolled steroid-naïve, symptomatic asthmatics with baseline exhaled nitric oxide (eNO) of ≥ 40 ppb. There were 120 eligible patients who were randomized and received a dose of inhaled budesonide/formoterol 320/9 μg (lower dose budesonide/formoterol), 640/18 μg (higher dose budesonide/formoterol (HDBF)), or terbutaline (TERB) 1 mg. Inflammatory cells and mediators in induced sputum, eNO and lung function were measured at baseline and 6 h (acute phase). Subsequently, all patients used BFMRT as real-life practice for 24 weeks (maintenance phase).. In the acute phase, the degree of post-treatment reduction in total eosinophil counts, interleukin-8 and matrix metalloproteinase-9 in induced sputum were significantly greater in group HDBF (vs TERB, P < 0.05). The increase in forced expiratory volume in first second (FEV1 ) in group HDBF was significantly higher (vs LDBF and TERB, P < 0.05) 3 h after dosing. In the maintenance phase, significant improvement of asthma control (presented by eNO, FEV1 and a five-item asthma control questionnaire) in real-life settings was observed at 4 weeks and sustained to the end of study. The rate of patients who followed scheduled visits declined over time (87% at week 4 and 42% at week 24).. Budesonide/formoterol as reliever exerts acute, dose-related anti-inflammatory effects and FEV1 improvement in symptomatic asthmatics. BFMRT is effective in asthma control. However, the decrease in long-term follow-up rate remains an issue to overcome in real-life settings.

Topics: Administration, Inhalation; Adult; Asthma; Breath Tests; Bronchodilator Agents; Budesonide, Formoterol Fumarate Drug Combination; Cell Count; Drug Combinations; Eosinophils; Ethanolamines; Exhalation; Female; Forced Expiratory Volume; Humans; Interleukin-8; Maintenance Chemotherapy; Male; Matrix Metalloproteinase 9; Middle Aged; Nitric Oxide; Sputum; Terbutaline; Vital Capacity; Young Adult

Women in poor urban neighborhoods have high rates of stress and allergic diseases, but whether stress or stress correlates such as depression promote inflammatory and type 2 cytokine responses is unknown.. To examine associations among external stressors, perceived stress, depression, and peripheral blood mononuclear cell cytokine responses of mothers enrolled in the Urban Environment and Childhood Asthma Study and test the hypothesis that stress would be positively associated with type 2 and selected proinflammatory (tumor necrosis factor-α and interleukin-8) responses.. Questionnaire data from mothers living in 4 inner cities included information about external stress, stress perception, and depression. The external stress domains (interpersonal problems, housing, and neighborhood stress) were combined into a Composite Stressor score. Peripheral blood mononuclear cells were stimulated ex vivo and cytokine responses to innate, adaptive, and polyclonal immune stimuli were compared with stress and depression scores for 469 of the 606 study participants.. There were no significant positive associations between Composite Stressor scores, perceived stress, or depression scores and proinflammatory or type 2 cytokine responses, and these findings were not modified by allergy or asthma status. There were some modest associations with individual stressors and cytokine responses, but no consistent relations were noted. Depression was associated with decreased responses to some stimuli, particularly dust mite.. Composite measurements of stressors, perceived stress, or depression were not positively related to proinflammatory or type 2 cytokine responses in these young urban women. These data do not support the hypothesis that these factors promote cytokine responses associated with allergy.. ClinicalTrials.gov, identifier NCT00114881.

Topics: Adolescent; Adult; Asthma; Cytokines; Depression; Female; Humans; Hypersensitivity; Interleukin-8; Leukocytes, Mononuclear; Mothers; Residence Characteristics; Stress, Psychological; Tumor Necrosis Factor-alpha; Urban Population; Young Adult

Selectins, a family of cell adhesion molecules, are involved in leukocyte extravasation to sites of inflammation. We investigated the safety and efficacy of the inhaled pan-selectin antagonist Bimosiamose in patients with chronic obstructive pulmonary disease (COPD). 77 COPD patients (mean forced expiratory volume in 1 s, 57% pred.) were enrolled in a cross-over, double-blind, randomized, Placebo-controlled, multi-center trial. Bimosiamose (10 mg) or Placebo was inhaled twice daily via the breath actuated nebulizer Akita2 Apixneb™ for 28 days on top of standard bronchodilator therapy. Efficacy was assessed by measurement of inflammatory parameters in induced sputum (differential cell count, interleukin-8, matrix-metalloproteinase-9, myeloperoxidase) and lung function at day 28 of both treatment periods. The total adverse event ratio of Bimosiamose compared to Placebo treatment was balanced. Compared to Placebo, treatment with Bimosiamose led to a decrease of the interleukin-8 concentration (-9.49 ng/mL, 95%CI -18.8 to -2.7 ng/mL, p = 0.008), for the neutrophil count a difference of -0.368 × 10(6) cells/mL (95%CI -1.256 to 0.407 × 10(6)/mL, p = 0.313) was found. The macrophage count decreased by -0.200 × 10(6) cells/mL (95%CI -0.365 to -0.044 × 10(6) cells/mL, p = 0.012). Most lung function parameters showed a small numeric increase. Inhalation of Bimosiamose for 28 days was safe and well tolerated in patients with COPD. It led to an attenuation of airway inflammation (EudraCT 2009-017257-35; NCT ID: NCT01108913).

Topics: Administration, Inhalation; Aged; Asthma; Cross-Over Studies; Double-Blind Method; Female; Hexanes; Humans; Interleukin-8; Male; Mannose; Matrix Metalloproteinase 9; Middle Aged; Pulmonary Disease, Chronic Obstructive; Selectins

Increased numbers of neutrophils are reported in the airways of patients with severe asthma. It is not clear if they contribute to the lack of control and severity. There are currently no strategies to investigate this by decreasing neutrophil numbers in the airways.. To investigate the safety and efficacy of SCH527123, a selective CXCR2 receptor antagonist, in patients with severe asthma and increased number of neutrophils in sputum.. In a randomized, double-blind, parallel study, patients with severe asthma and sputum total cell count < 10 × 10(6) /g and neutrophils > 40% were randomized to SCH527123, 30 mg daily PO (n = 22) or placebo (n = 12) for 4 weeks. Primary end-points were safety and change in sputum and blood neutrophil counts. Secondary end-points were change in asthma control questionnaire (ACQ) score, minor and major exacerbations, spirometry and sputum neutrophil activation markers.. The SCH 527123 caused a mean reduction of 36.3% in sputum neutrophil percentage compared to a 6.7% increase in the placebo arm (P = 0.03). The mean absolute neutrophil count in blood was reduced by 14% at the end of 4 weeks, but recovered by the 5th week. There were no differences in the overall rates of adverse events among the groups. There were fewer mild exacerbations (1.3 vs. 2.25, P = 0.05) and a trend towards improvement in the ACQ score (mean difference between groups of 0.42 points, P = 0.053). No statistically significant changes were observed in forced expiratory volume in 1 s (FEV (1)), sputum myeloperoxidase, IL8 or elastase.. The SCH527123 is safe and reduces sputum neutrophils in patients with severe asthma.. This new treatment provides an opportunity to investigate the role of neutrophils in severe asthma with potential clinical benefits. Larger studies of longer duration are needed to evaluate the impact on other outcomes of asthma including exacerbations.

Topics: Adolescent; Adult; Aged; Asthma; Benzamides; Biomarkers; Cell Count; Cyclobutanes; Double-Blind Method; Female; Humans; Interleukin-8; Male; Middle Aged; Neutrophil Activation; Neutrophils; Pancreatic Elastase; Peroxidase; Receptors, Interleukin-8B; Severity of Illness Index; Sputum

Combination therapy with inhaled corticosteroids (ICS) and long-acting β(2)-adrenergic agonists (LABA) is reported to have superior effects on controlling asthma symptoms to ICS alone; however, there is no molecular-based evidence to explain the clinical effects. Here, the effect of the ICS/LABA combination was compared with ICS on glucocorticoid receptor (GR) activation in sputum macrophages.. In a randomised, double-blind cross-over placebo-controlled 6-visit study, 10 patients with mild asthma were given placebo, formoterol (Oxis(®) 12 μg), budesonide (Pulmicort(®) 200 μg :BUD200, or 800 μg :BUD800), or budesonide/formoterol combination (Symbicort(®)) as a single 100/6 μg (SYM100) or double 200/12 μg (SYM200) dose. Sputum macrophages were separated by plate adhesion from induced sputum. GR binding to the glucocorticoid-response elements on oligonucleotides (GR-GRE binding) was evaluated by ELISA. mRNA expression of MAP-kinase phosphatase (MKP)-1 and IL-8 were measured by quantitative RT-PCR.. GR-GRE binding was significantly increased after treatment with SYM100 (3.5 OD/10 μg protein, median, p < 0.05) versus placebo (1.3) and BUD200 (1.6), and the induction was higher than that of BUD800 (2.4). MKP-1 mRNA was increased and IL-8 mRNA was significantly inhibited by BUD800, SYM100 and SYM200 versus placebo.. The effects of SYM100 and SYM200 on GR activation were not different from that of BUD800 and superior to BUD200. Thus, it has been confirmed at a molecular level that inhaled combination therapy with a lower dose of budesonide has an equivalent effect to a high dose of budesonide alone. In addition, GR-GRE binding is found to be a valuable pharmacodynamic marker for steroid efficacy in clinical studies.

Topics: Administration, Inhalation; Adrenal Cortex Hormones; Adult; Anti-Asthmatic Agents; Asthma; Biomarkers; Bronchodilator Agents; Budesonide; Budesonide, Formoterol Fumarate Drug Combination; Cross-Over Studies; Dose-Response Relationship, Drug; Double-Blind Method; Drug Combinations; Drug Therapy, Combination; Dual Specificity Phosphatase 1; Enzyme-Linked Immunosorbent Assay; Ethanolamines; Female; Glucocorticoids; Humans; Interleukin-8; Male; Response Elements; Reverse Transcriptase Polymerase Chain Reaction; Sputum

Severe asthma occurs in a heterogeneous group of patients in whom symptoms and airway inflammation persist despite maximal antiasthma treatment.. To verify whether a short-term course of oral steroids would modify sputum inflammatory cytokine and sputum eosinophil concentrations and whether this effect is related to the presence of sputum eosinophilia.. In 59 patients with severe refractory asthma, we measured pulmonary function and inflammatory markers in hypertonic saline-induced sputum before and after 2 weeks of treatment with 0.5 mg/kg of oral prednisone (n = 39) or placebo (n = 20) daily. Selected sputum portions were assayed for total and differential cell counts and supernatant interleukin (IL) 5 and IL-8 concentrations.. At baseline, no statistical differences were found among placebo- and prednisone-treated patients in terms of sputum inflammatory cell percentages and IL-5 and IL-8 concentrations. After treatment, forced expiratory volume in 1 second significantly increased and sputum eosinophil percentages and IL-5 and IL-8 concentrations significantly decreased in the prednisone group, whereas no changes were observed in the placebo group. The positive effect of prednisone treatment was observed only in patients with baseline sputum eosinophilia, whereas in noneosinophilic patients with severe asthma prednisone induced only a significant decrease of sputum IL-8.. Additional high-dose oral corticosteroids improve pulmonary function and reduce not only sputum eosinophil but also sputum proinflammatory cytokine concentrations in patients with severe refractory asthma.

Topics: Administration, Oral; Adult; Aged; Asthma; Double-Blind Method; Eosinophils; Female; Humans; Interleukin-5; Interleukin-8; Leukocyte Count; Male; Middle Aged; Prednisone; Sputum

About 5-10% of patients with asthma suffer from poorly-controlled disease despite corticosteroid (CS) therapy.. We determined whether there were any differences in inflammatory biomarkers between severe and non-severe asthma patients.. Nineteen severe and 20 non-severe asthma patients were recruited and underwent collection of induced sputum, bronchoalveolar lavage (BAL) fluid and bronchial biopsies.. Biopsy results showed no differences in eosinophils (major basic protein positive), neutrophils, macrophages, T cells and mast cells in the bronchial submucosa. However, subbasement membrane (SBM) thickness and smooth muscle area were increased in the biopsies. No significant differences were observed in the induced sputum inflammatory cells. In BAL fluid, there was a significant increase in neutrophils but a significant decrease in macrophages. Eosinophil counts were non-significantly increased threefold in both sputum and BAL in severe asthma. Levels of IL-8 and IL-13 in sputum supernatants were similar in both groups of asthma patients. There was a significant inverse correlation between post-bronchodilator forced expiratory volume in 1 s and provocative concentration of methacholine causing a 20% fall in FEV(1) with SBM thickness.. Differences in inflammatory cells were observed mainly in terms of increased neutrophils and reduction in macrophage numbers in BAL fluid with a trend towards increased eosinophils in severe asthma compared with non-severe asthma. However, the most notable features are the increase in features of airway wall remodelling of SBM thickness and smooth muscle area.

Topics: Adult; Asthma; Biomarkers; Female; Humans; Inflammation; Inflammation Mediators; Interleukin-13; Interleukin-8; Leukocytes; Male; Respiratory Mucosa; Severity of Illness Index; Sputum

Patients with refractory asthma have persistent symptoms despite maximal treatment with inhaled corticosteroids and long-acting bronchodilators. The availability of add-on therapies is limited, and effective add-on therapies that target noneosinophilic airway inflammation are needed. Macrolide antibiotics, such as clarithromycin, have in vitro efficacy against IL-8 and neutrophils, key inflammatory mediators in noneosinophilic asthma.. To determine the efficacy of clarithromycin in patients with severe refractory asthma and specifically in a subgroup of patients with noneosinophilic asthma.. Subjects with severe refractory asthma (n = 45) were randomized to receive clarithromycin (500 mg twice daily) or placebo for 8 weeks.. The primary outcome for this study was sputum IL-8 concentration. Other inflammatory outcomes assessed included sputum neutrophil numbers and concentrations of neutrophil elastase and matrix metalloproteinase (MMP)-9. Clinical outcomes were also assessed, including lung function, airway hyperresponsiveness to hypertonic saline, asthma control, quality of life, and symptoms. Clarithromycin therapy significantly reduced airway concentrations of IL-8 and neutrophil numbers and improved quality-of-life scores compared with placebo. Reductions in neutrophil elastase and MMP-9 concentrations were also observed. These reductions in inflammation were most marked in those with refractory noneosinophilic asthma.. Clarithromycin therapy can modulate IL-8 levels and neutrophil accumulation and activation in the airways of patients with refractory asthma. Macrolide therapy may be an important additional therapy that could be used to reduce noneosinophilic airway inflammation, particularly neutrophilic inflammation, in asthma. Clinical trial registered with the Australian Clinical Trials Registry www.actr.org.au (No. 12605000318684).

Topics: Adult; Aged; Aged, 80 and over; Anti-Bacterial Agents; Asthma; Clarithromycin; Double-Blind Method; Drug Therapy, Combination; Female; Gene Expression; Humans; Interleukin-8; Leukocyte Count; Male; Middle Aged; Neutrophils; Quality of Life; Sputum

The role of neutrophils in exacerbations of asthma is poorly understood. We examined the effect of withdrawal of inhaled corticosteroids on sputum inflammatory indexes in a double-blind study in patients with moderate, stable asthma.. Following a 2-week run in period, 24 subjects were randomized to receive either budesonide (400 microg bid) or placebo, and the study was continued for another 10 weeks.. Loss of asthma control developed in 8 of 12 patients over the 10-week period of steroid withdrawal, whereas only 1 of 10 patients with budesonide treatment had exacerbations. Those with an exacerbation had increased sputum interleukin (IL)-8 (p < 0.0001) and increased sputum neutrophil numbers (p < 0.0001) compared to those without an exacerbation. The significant elevation in sputum IL-8 and neutrophil counts initially occurred 2 weeks prior to an exacerbation. Sputum neutrophilia correlated positively with changes in IL-8 levels (r(2) = 0.76, p = 0.01).. Rapid withdrawal of inhaled corticosteroids results in an exacerbation of asthma that is preceded by an increase in sputum neutrophils and IL-8 concentrations, in contrast to an increase in eosinophils reported in previous studies in which inhaled steroids are slowly tapered.

Topics: Adolescent; Adult; Asthma; Budesonide; Cell Count; Double-Blind Method; Eosinophils; Female; Glucocorticoids; Humans; Interleukin-8; Male; Middle Aged; Neutrophils; Sputum

Few studies have compared treatment strategies in patients with asthma poorly controlled on low dose inhaled corticosteroids, and little is known about the effects of different treatments on airway inflammation. In this double-blind, placebo-controlled, parallel group study, we compared the effects of salmeterol plus fluticasone propionate (FP) (Seretide; SFC) and FP plus montelukast (FP/M) on sputum inflammatory markers, airway responsiveness, lung function, and symptoms in adult asthmatics.. Sixty-six subjects were randomised to SFC or FP/M for 12 weeks. The primary outcome was changes in neutrophil, eosinophil, macrophage, lymphocyte, and epithelial cell levels in induced sputum. Additional outcomes included the change in other sputum markers of airway inflammation, airway responsiveness, symptom control, and lung function.. Both treatments had no significant effect on induced sputum inflammatory cells, although there was a trend for a reduction in sputum eosinophils. Both treatments significantly improved airway responsiveness, whereas SFC generally led to greater improvements in symptom control and lung function than FP/M. FP/M led to significantly greater reductions in sputum cysteinyl leukotrienes than SFC (treatment ratio 1.80; 95% CI 1.09, 2.94).. Both treatments led to similar control of eosinophilic airway inflammation, although PEF and symptom control were better with SFC. STUDY NUMBER: SAM40030 (SOLTA).

Topics: Acetates; Adrenergic beta-Agonists; Adult; Albuterol; Androstadienes; Anti-Asthmatic Agents; Asthma; Bronchial Hyperreactivity; Cyclopropanes; Cysteine; Double-Blind Method; Drug Combinations; Female; Fluticasone; Fluticasone-Salmeterol Drug Combination; Forced Expiratory Volume; Histamine; Humans; Interleukin-8; Leukotriene Antagonists; Leukotrienes; Lung; Male; Quinolines; Spirometry; Sputum; Sulfides; Time Factors; Treatment Outcome; United Kingdom

The role of Th2 cells (producing interleukin (IL-)4, IL-5 and IL-13) in allergic asthma is well-defined. A distinct proinflammatory T cell lineage has recently been identified, called Th17 cells, producing IL-17A, a cytokine that induces CXCL8 (IL-8) and recruits neutrophils. Neutrophilic infiltration in the airways is prominent in severe asthma exacerbations and may contribute to airway gland hypersecretion, bronchial hyper-reactivity and airway wall remodelling in asthma.. to study the production of IL-17 in asthmatic airways at the mRNA level, and to correlate this with IL-8 mRNA, neutrophilic inflammation and asthma severity.. We obtained airway cells by sputum induction from healthy individuals (n = 15) and from asthmatic patients (n = 39). Neutrophils were counted on cytospins and IL-17A and IL-8 mRNA expression was quantified by real-time RT-PCR (n = 11 controls and 33 asthmatics).. Sputum IL-17A and IL-8 mRNA levels are significantly elevated in asthma patients compared to healthy controls. IL-17 mRNA levels are significantly correlated with CD3gamma mRNA levels in asthmatic patients and mRNA levels of IL-17A and IL-8 correlated with each other and with sputum neutrophil counts. High sputum IL-8 and IL-17A mRNA levels were also found in moderate-to-severe (persistent) asthmatics on inhaled steroid treatment.. The data suggest that Th17 cell infiltration in asthmatic airways links T cell activity with neutrophilic inflammation in asthma.

Topics: Adolescent; Adult; Aged; Asthma; Female; Granulocytes; Humans; Interleukin-17; Interleukin-8; Male; Middle Aged; RNA, Messenger; Sputum; T-Lymphocytes

Nocturnal airway obstruction occurs frequently in childhood asthma and results from increased airway inflammation. Lymphocytes are believed to be key effector cells of airway wall inflammation, releasing pro-inflammatory mediators and cytokines. A previous study showed that hydrocortisone infusion, an effective anti-inflammatory treatment, improves nocturnal and daytime FEV(1) values. This study in 16 children with moderate asthma was designed to assess whether there exists day and night differences in IL-4, IL-5, IL-8, and IFN-gamma production of concanavaline A stimulated peripheral blood mononuclear cells. Furthermore, we investigated whether substitution of low serum cortisol levels with intravenous hydrocortisone would affect those parameters. Saline (as a placebo) or hydrocortisone (30 microg/m(2) body surface area/24 h) was intravenously administered in a randomized, double blind, cross-over design. Measurements under saline or hydrocortisone infusions were separated by 1 wk. At 04:00 and 16:00 hours 10 ml blood was taken for determination of peripheral blood mononuclear cell isolation and stimulation, and an eosinophil count. Hydrocortisone infusion significantly reduced the nocturnal fall in FEV(1). Median values of IFNgamma, IL-4, IL-5, and IL-8 produced by peripheral blood mononuclear cells did not significantly differ at 04:00 and 16:00 hours, both with saline and hydrocortisone infusion. Our results suggest that FEV(1) improvement is not due to suppression of circulating peripheral blood mononuclear cell activation. We hypothesize that it is rather due to its effect on local lung tissue epithelial and/or fibroblasts thereby reducing airway inflammation and vascular leakage.

Topics: Adolescent; Airway Obstruction; Anti-Inflammatory Agents; Asthma; Child; Circadian Rhythm; Concanavalin A; Cross-Over Studies; Cytokines; Double-Blind Method; Female; Forced Expiratory Volume; Humans; Hydrocortisone; Infusions, Intravenous; Interferon-gamma; Interleukin-4; Interleukin-5; Interleukin-8; Leukocytes, Mononuclear; Male; Time Factors

Airway hyperresponsiveness and inflammation can be noninvasively studied by bronchial provocation using direct (histamine) or indirect (adenosine 5'-monophosphate [AMP]) stimuli and induced sputum.. To report on the immediate effects of histamine and AMP challenge on induced sputum neutrophil counts and related mediator levels.. We performed a single-masked, randomized, placebo-controlled, 3-way, crossover, methodological study in 14 atopic patients (median age, 25 years; 8 males; mean +/- SD forced expiratory volume in 1 second, 99% +/- 5%) without anti-inflammatory medication use. At baseline, sputum induction was performed. Bronchial challenges with AMP, histamine, or placebo were performed 48 hours later. Thirty minutes after challenge, sputum induction was performed again. Challenge periods in each patient were separated by more than 2 weeks. Sputum cells and the mediators leukotriene B4, interleukin 8, myeloperoxidase, and albumin were quantified.. Comparing median challenge-induced relative changes in cells and mediators, neither histamine nor AMP challenge altered the induced sputum neutrophil counts (histamine, 2.7%; AMP, 2.95%; placebo, -2%; P > .07 for all), interleukin 8 levels (histamine, 2.4 ng/mL; AMP, -3.8 ng/mL; placebo, -0.2 ng/mL; P > .06), leukotriene B4 levels (histamine, -4.8 pg/mL; AMP, 3 pg/mL; placebo, 6 pg/mL; P > .08), or myeloperoxidase levels (histamine, 0.16 microg/mL; AMP, 0 microg/mL; placebo, -0.03 microg/mL; P > .07). Sputum albumin levels were increased after histamine challenge compared with AMP and placebo challenge (P < .01 for both).. Histamine and AMP provocation have no major effects on induced neutrophil counts and related mediator levels in atopic patients, whereas histamine challenge induces plasma leakage. Potential interactions of noninvasive methods to evaluate airway reactivity and inflammation should be carefully considered.

Topics: Adenosine Monophosphate; Adult; Asthma; Bronchial Provocation Tests; Cell Count; Cross-Over Studies; Female; Histamine; Humans; Interleukin-8; Leukotriene B4; Male; Neutrophils; Peroxidase; Rhinitis, Allergic, Seasonal; Sputum

Particulate matter (PM) pollution adversely affects the airways, with asthmatic subjects thought to be especially sensitive. The authors hypothesised that exposure to diesel exhaust (DE), a major source of PM, would induce airway neutrophilia in healthy subjects, and that either these responses would be exaggerated in subjects with mild allergic asthma, or DE would exacerbate pre-existent allergic airways. Healthy and mild asthmatic subjects were exposed for 2 h to ambient levels of DE (particles with a 50% cut-off aerodynamic diameter of 10 microm (PM10) 108 microg x m(-3)) and lung function and airway inflammation were assessed. Both groups showed an increase in airway resistance of similar magnitude after DE exposure. Healthy subjects developed airway inflammation 6 h after DE exposure, with airways neutrophilia and lymphocytosis together with an increase in interleukin-8 (IL-8) protein in lavage fluid, increased IL-8 messenger ribonucleic acid expression in the bronchial mucosa and upregulation of the endothelial adhesion molecules. In asthmatic subjects, DE exposure did not induce a neutrophilic response or exacerbate their pre-existing eosinophilic airway inflammation. Epithelial staining for the cytokine IL-10 was increased after DE in the asthmatic group. Differential effects on the airways of healthy subjects and asthmatics of particles with a 50% cut-off aerodynamic diameter of 10 microm at concentrations below current World Health Organisation air quality standards have been observed in this study. Further work is required to elucidate the significance of these differential responses.

Topics: Adult; Airway Resistance; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Cell Adhesion Molecules; Environmental Exposure; Female; Humans; Inflammation; Interleukin-10; Interleukin-8; Lymphocytosis; Male; Middle Aged; Neutrophils; Respiratory Mucosa; Respiratory System; RNA, Messenger; Vehicle Emissions

The inflammatory response to ozone in atopic asthma suggests that soluble mediators of inflammation are released in response to oxidant stress. Antioxidants may alleviate additional oxidative stress associated with photochemical oxidant pollution. This study investigates the impact of antioxidant supplementation on the nasal inflammatory response to ozone exposure in atopic asthmatic children. We conducted a randomized trial using a double-blinded design. Children with asthma (n = 117), residents of Mexico City, were given randomly a daily supplement of vitamins (50 mg/day of vitamin E and 250 mg/day of vitamin C) or placebo. Nasal lavages were performed three times during the 4-month follow-up and analysed for content of interleukin-6 (IL-6), IL-8, uric acid and glutathione (GSx). IL-6 levels in the nasal lavage were increased significantly in the placebo group after ozone exposure while no increase was observed in the supplement group. The difference in response to ozone exposure between the two groups was significant (P = 0.02). Results were similar for IL-8, but with no significant difference between the groups (P = 0.12). GSx decreased significantly in both groups. Uric acid decreased slightly in the placebo group. Our data suggest that vitamin C and E supplementation above the minimum dietary requirement in asthmatic children with a low intake of vitamin E might provide some protection against the nasal acute inflammatory response to ozone.

Topics: Air Pollutants; alpha-Tocopherol; Antioxidants; Ascorbic Acid; Asthma; Child; Dietary Supplements; Double-Blind Method; Environmental Exposure; Female; Glutathione; Humans; Interleukin-6; Interleukin-8; Male; Nasal Cavity; Oxidative Stress; Ozone; Uric Acid; Vitamin E

The aqueous extract of propolis has been formulated as a nutritional food product and administered, as an adjuvant to therapy, to patients with mild to moderate asthma daily for 2 months in the framework of a comparative clinical study in parallel with a placebo preparation. The diagnosis of asthma was made according to the criteria of patient classification of the National Institutes of Health and Global Initiative for Asthma Management. At inclusion, the pulmonary forced expiratory volume in the first second (FEV1) as a percentage of the forced vital capacity (FVC) was more than 80% in mild persistent cases, and between 60 and 80% in moderate persistent cases, showing an increase in the degree of reversibility of > 15% in FEV1. All patients were on oral theophylline as controller therapy, none was receiving oral or inhaled corticosteroids, none had other comorbidities necessitating medical treatment, and all were from a middle-class community and had suffered from asthma for the last 2-5 years. Twenty-four patients received the placebo, with one drop-out during the study, while 22 received the propolis extract, with no drop-outs. The age range of the patients was 19-52 years; 36 were male and 10 female. The number of nocturnal attacks was recorded on a weekly basis, while pulmonary function tests were performed on all patients at the beginning of the trial, 1 month later and at the termination of the trial. Immunological parameters, including various cytokines and eicosanoids known to play a role in asthma, were measured in all patients at the beginning of the trial and 2 months later. Analysis of the results at the end of the clinical study revealed that patients receiving propolis showed a marked reduction in the incidence and severity of nocturnal attacks and improvement of ventilatory functions. The number of nocturnal attacks dropped from an average of 2.5 attacks per week to only 1. The improvement in pulmonary functions was manifested as a nearly 19% increase in FVC, a 29.5% increase in FEV1, a 30% increase in peak expiratory flow rate (PEFR), and a 41% increase in the forced expiratory flow rate between 25 and 75% of the vital capacity (FEF25-75). The clinical improvement was associated with decreases by 52, 65, 44 and 30%, respectively, of initial values for the pro-inflammatory cytokines tumor necrosis factor (TNF)-alpha, ICAM-1, interleukin (IL)-6 and IL-8, and a 3-fold increase in the 'protective' cytokine IL-10. The levels of prostaglandins

Topics: Adult; Animals; Anti-Inflammatory Agents; Asthma; Dietary Supplements; Female; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Leukotrienes; Male; Medicine, Traditional; Middle Aged; Milk; Propolis; Respiratory Function Tests

In-vitro data suggest that long-acting beta2-agonists may have a neutrophil-stabilising effect. A reduction in airway wall eosinophil number following introduction of salmeterol in persistent asthma has previously been described. There is currently interest in the "neutrophil system" in asthma, and thus the aim of the present study was to investigate the effect of salmeterol on interleukin (IL)-8, neutrophils and myeloperoxidase (MPO) in persistent asthma. In the same 12-week double-blind parallel-group placebo-controlled study as described previously, the effects on bronchoalveolar lavage fluid (BALF) IL-8, neutrophils and MPO of introducing salmeterol (50 microg b.i.d.) or giving additional inhaled corticosteroid (fluticasone 100 microg b.i.d.) in 45 subjects with persistent asthma already on low/moderate doses of inhaled corticosteroids were further investigated. At baseline, BALF IL-8 but not neutrophil or MPO levels were significantly raised in the asthmatic subjects compared to normal controls. MPO levels correlated strongly with IL-8 levels, and weakly with BALF neutrophil numbers in the asthmatics. Fluticasone treatment resulted in significantly elevated neutrophil numbers, but not MPO or IL-8 levels. In contrast, introducing salmeterol significantly reduced IL-8 and MPO levels, but did not affect BALF neutrophil numbers. Interestingly, salmeterol and fluticasone showed significantly contrasting effects on MPO and neutrophils, and there was a divergent effect on IL-8 levels that almost reached significance. Excessive interleukin-8 levels may be relevant to asthma pathogenesis, even in the setting of moderate-dose inhaled corticosteroid therapy. Reduction in interleukin-8 production and possibly stabilisation of airway neutrophil numbers may explain the greater clinical benefit of adding a long-acting beta2-agonist rather than merely increasing inhaled corticosteroid doses. Indeed, high-dose inhaled corticosteroid therapy alone may promote airway neutrophilia.

Topics: Adrenergic beta-Agonists; Adult; Aged; Albuterol; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Bronchoscopy; Dose-Response Relationship, Drug; Double-Blind Method; Female; Humans; Interleukin-8; Male; Middle Aged; Neutrophil Activation; Peroxidase; Salmeterol Xinafoate; Time Factors

Ozone (O3) is a common air pollutant associated with adverse health effects. Asthmatics have been suggested to be a particularly sensitive group.. This study evaluated whether bronchial epithelial cytokine expression would differ between healthy and allergic asthmatics after ozone exposure, representing an explanatory model for differences in susceptibility.. Healthy and mild allergic asthmatic subjects (using only inhaled beta2-agonists prn) were exposed for 2 h in blinded and randomized sequence to 0.2 ppm of O3 and filtered air. Bronchoscopy with bronchial mucosal biopsies was performed 6 h after exposure. Biopsies were embedded in GMA and stained with mAbs for epithelial expression of IL-4, IL-5, IL-6, IL-8, IL-10, TNF-alpha, GRO-alpha, granulocyte-macrophage colony-stimulating factor (GM-CSF), fractalkine and ENA-78.. When comparing the two groups at baseline, the asthmatic subjects showed a significantly higher expression of IL-4 and IL-5. After O3 exposure the epithelial expression of IL-5, GM-CSF, ENA-78 and IL-8 increased significantly in asthmatics, as compared to healthy subjects.. The present study confirms a difference in epithelial cytokine expression between mild atopic asthmatics and healthy controls, as well as a differential epithelial cytokine response to O3. This O3-induced upregulation of T helper type 2 (Th2)-related cytokines and neutrophil chemoattractants shown in the asthmatic group may contribute to a subsequent worsening of the airway inflammation, and help to explain their differential sensitivity to O3 pollution episodes.

Topics: Adult; Air Pollutants; Asthma; Bronchi; Case-Control Studies; Chemokine CXCL5; Chemokines, CXC; Cytokines; Disease Susceptibility; Epithelium; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunohistochemistry; Interleukin-4; Interleukin-5; Interleukin-8; Male; Ozone

Regular salbutamol use can exacerbate allergen-induced airway eosinophilia in asthmatics, but its effect on airway eosinophil chemokine responses is unknown. Asthmatic subjects (n=14) were treated for 10 days with placebo or salbutamol in a double-blind, cross-over study, then given same-dose allergen challenges. Their sputa were then analysed 1 and 7 h later for a panel of eosinophil-related cytokines. Eosinophils from five test and three control subjects were tested for expression of CXCL8/interleukin (IL)-8, and its receptors and responsiveness to CCL11/eotaxin and CXCL8/IL-8. Sputum CXCL8/IL-8, but not IL-5, CCL5/regulated on activation, T-cell expressed and secreted, CCL7/monocyte chemotactic protein-3, CCL11/eotaxin, granulocyte-macrophage colony-stimulating factor or tumour necrosis factor levels, were increased (42%) by the salbutamol treatments. The CXCL8/IL-8 levels correlated with the proportions of sputum eosinophils and these cells, but not other sputum cells, stained strongly for CXCL8/IL-8. The circulating eosinophils of the tested subjects (n=5) expressed CXCL8/IL-8 receptors and secreted high levels of this chemokine. Neutralisation of sputum CXCL8/IL-8 reduced eosinophil chemotactic responses to these samples by 19 +/- 5%. These data suggest that regular use of salbutamol can augment airway CXCL8/interleukin-8 responses to allergen challenge and that this CXCL8/interleukin-8 could contribute to the airway inflammatory response.

Topics: Adult; Albuterol; Analysis of Variance; Asthma; Bronchial Provocation Tests; Bronchodilator Agents; Chemokines, CXC; Cross-Over Studies; Double-Blind Method; Enzyme-Linked Immunosorbent Assay; Eosinophils; Female; Humans; Immunohistochemistry; Inflammation Mediators; Interleukin-8; Male; Middle Aged; Sputum; Statistics, Nonparametric; Up-Regulation

Exacerbations of asthma are often associated with rhinovirus (RV)-induced common colds. During experimental RV-infection in healthy subjects, increased levels of the pro-inflammatory mediator IL-1beta and the anti-inflammatory IL-1 receptor antagonist (IL-1ra) have been found in nasal lavage.. We postulated that the balance between nasal pro- and anti-inflammatory mediator expression is disturbed in asthma, resulting in more extensive inflammation following RV-exposure in asthma.. We determined IL-1ra, IL-1beta, and IL-8 in nasal lavages (days -2, 3, and 6) of non-asthmatics and asthmatics (with and without pre-treatment with the inhaled steroid budesonide) before and after experimental RV16-infection (days 0 and 1).. Following RV16-infection, a significant increase in IL-8 was observed in the placebo- and budesonide-treated asthmatics (P=0.033 and 0.037, respectively), whereas IL-1beta only increased in the two asthma groups combined (P=0.035). A small, but significant, increase in IL-1ra was only observed in the budesonide-treated asthmatics (P=0.047). At baseline, IL-1ra levels were significantly higher in the non-asthmatics than in the placebo-treated asthmatics (P=0.017).. These results demonstrate differences between non-asthmatic and asthmatic subjects in the basal levels of nasal cytokines and their inhibitors, and in the effect of experimental RV-infection on these levels. The results indicate that RV may enhance inflammation more markedly in asthmatics, and suggest that this may in part be explained by lower IL-1ra levels. In addition, the observation that budesonide-treatment may result in higher nasal IL-1ra levels supports the hypothesis that steroids act in part by increasing the endogenous anti-inflammatory screen.

Topics: Asthma; Bronchodilator Agents; Budesonide; Common Cold; Double-Blind Method; Humans; Inflammation Mediators; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Interleukin-8; Nasal Lavage Fluid; Rhinovirus; Sialoglycoproteins

We investigated whether exposure to ozone (O(3)) 24 hours after an allergen challenge test would increase airway eosinophilia induced by allergen in subjects with mild asthma with late airway response. Twelve subjects with mild atopic asthma participated in a randomized, single-blind study. Subjects underwent allergen challenge 24 hours before a 2 hour exposure to O(3) (0.27 ppm) or filtered air. Pulmonary function was monitored during the allergen challenge and after the exposure to O(3) or air. Six hours later, induced sputum was collected. After 4 weeks, the experiment was repeated with the same subjects. Allergen induced a comparable late airway response in both challenges. O(3) exposure induced a significant decrease in FVC, FEV(1), and vital capacity, and was associated with a significant increase in total symptom score compared with air exposure. The percentage of eosinophils, but not the percentage of neutrophils, in induced sputum was significantly higher after exposure to O(3) than after exposure to air (p = 0.04). These results indicate that O(3) exposure after a late airway response elicited by allergen challenge can potentiate the eosinophilic inflammatory response induced by the allergen challenge itself in subjects with mild atopic asthma. This observation may help explain the synergistic effect of air pollution and allergen exposure in the exacerbation of asthma.

Topics: Adolescent; Adult; Allergens; Asthma; Bronchial Provocation Tests; Bronchoconstrictor Agents; Eosinophils; Forced Expiratory Volume; Humans; Interleukin-8; Leukocyte Count; Methacholine Chloride; Oxidants, Photochemical; Ozone; Respiratory Mechanics; Single-Blind Method; Sputum; Vital Capacity

A knowledge of the factors that can affect induced sputum results is essential in order to standardize the procedure.. We investigated the influence of nebulizer output on sputum cell counts and fluid phase measurements at increasing times of sputum induction.. Eighteen adults with stable asthma inhaled an aerosol of 3% hypertonic saline to induce sputum after 7, 14 and 21 min on 2 days separated by 48 h. On one day, in random order, the ultrasonic nebulizer used had a relatively low output of 0.87 mL/min (particle size 5.58 microm mass median aerodynamic diameter, MMAD) and, on the other, a higher output of 1.90 mL/min (particle size 4.14 microm MMAD). The sputum was selected from each expectorate and examined blind to the induction procedures.. With both nebulizers, the 14- and 21-min samples were lower in weight, neutrophils, eosinophils, eosinophil cationic protein (ECP) and interleukin (IL)-8 and higher in macrophages. The higher output nebulizer induced sputum with higher cell viability and lower ECP and IL-8.. The results identify that the volume of hypertonic saline inhaled in sputum induction influences the fluid-phase measurements. The duration of induction does alter the cell counts and suggests that the later expectorated sputum samples originate from more peripheral airways. The results draw attention to the need to standardize the volume and time of nebulization to accurately interpret and compare results.

Topics: Administration, Inhalation; Adult; Asthma; Blood Proteins; Cell Count; Cross-Over Studies; Cross-Sectional Studies; Eosinophil Granule Proteins; Eosinophils; Female; Forced Expiratory Volume; Humans; Inhalation Exposure; Interleukin-8; Macrophages; Male; Nebulizers and Vaporizers; Neutrophils; Ribonucleases; Saline Solution, Hypertonic; Sputum; Time Factors

Ambient ozone concentration is related to asthma exacerbation, but few findings are available regarding the effects of pharmacologic asthma treatment on this relationship. The purpose of this study was to investigate whether inhaled corticosteroids inhibit ozone-induced airway neutrophilic inflammation, as detected in induced sputum, and reduce functional response to ozone exposure. Eleven subjects with mild persistent asthma were exposed for 2 h, on separate days, to 0.27 ppm ozone and to air in random order, before and after 4 wk of treatment with budesonide (400 microg twice daily). Before exposure, 1 and 2 h after the beginning of exposure, and 6 h after the end of exposure, pulmonary function was measured, and a total symptom score questionnaire was completed; 6 h after exposure, sputum was induced with hypertonic saline. Budesonide treatment did not inhibit the functional response to ozone exposure, as determined by reduction in FEV(1) and increase in total symptom score, but it significantly blunted the increase in the percentage of sputum neutrophils and interleukin-8 concentrations in the supernatant (p < 0.05). Therefore, 4 wk of inhaled budesonide blunted the airway neutrophilic inflammatory response but did not prevent the functional impairment of the airways after ozone exposure.

Topics: Administration, Inhalation; Adult; Air Pollutants; Anti-Inflammatory Agents; Asthma; Bronchodilator Agents; Budesonide; Female; Forced Expiratory Volume; Humans; Interleukin-8; Male; Middle Aged; Neutrophils; Ozone; Respiratory Mechanics; Respiratory System; Single-Blind Method; Sputum; Vital Capacity

Recent epidemiologic and in vivo studies have suggested that inhaled endotoxin plays an important role in asthma pathogenesis.. The present study examines the effect of nasal allergen provocation on subsequent endotoxin challenge in subjects with atopic asthma.. By using a split-nose randomized crossover design, individual nares of 12 asthmatic subjects underwent challenge and lavage as follows. Immediately after a baseline nasal lavage, one nares received normal saline, and the other received dust mite antigen. Four hours later, both nares were exposed to either saline or endotoxin. Dust mite antigen (Dermatophagoides farinae) and endotoxin (Escherichia coli 026:B6) doses were 100 AU and 1000 ng, respectively. Postchallenge lavages were done at 8 and 24 hours after the initial challenge. The subjects then returned a minimum of 3 weeks later for crossover to the study arm. Nasal lavage fluid was analyzed for total and differential cell counts, IL-8, IL-6, intercellular adhesion molecule 1, GM-CSF, eosinophil cationic protein, myeloperoxidase, and soluble CD14.. A significant increase in the total inflammatory cell count was seen at 8 hours for the dust mite/endotoxin exposure compared with the saline/saline and saline/endotoxin exposures. Differential cell counts revealed a similar neutrophilic and eosinophilic inflammation for the dust mite/endotoxin exposure at 8 hours.. These data demonstrate an interaction between allergen and endotoxin exposure in asthmatic subjects, suggesting that a prior allergen challenge significantly augments the endotoxin-induced inflammation. Moreover, these data provide further evidence that concomitant exposure to allergen and endotoxin may be an important factor in asthma pathogenesis.

Topics: Adult; Asthma; Cross-Over Studies; Endotoxins; Female; Humans; Hypersensitivity, Immediate; Inflammation Mediators; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; Lipopolysaccharide Receptors; Male; Nasal Lavage Fluid; Nasal Provocation Tests; Rhinitis; Solubility

Topics: Administration, Inhalation; Adult; Androstadienes; Anti-Inflammatory Agents; Asthma; Female; Fluticasone; Humans; Interleukin-8; Male; Reference Values; Sensitivity and Specificity

Cytokine-mediated interactions among the inflammatory cells may play a role in the pathogenesis of bronchial asthma. Interleukin-8 (IL-8) is a major cytokine in the recruitment of neutrophils to the area of inflammation. Serum IL-8 is a marker of disease activity and treatment efficacy in bronchial asthma. To understand the role of IL-8 in disease activity in acute asthma, changes in serum concentrations of IL-8 elaborated by activated eosinophil before and after prednisolone therapy with clinical improvement were determined in the present study. Circulating levels of IL-8 in 15 normal control subjects and in sera from 20 allergic asthmatic children with acute exacerbation and in stable condition were determined by using commercially available assay kits. The mean concentration of serum IL-8 was statistically significantly higher in asthmatic children with acute exacerbation (63.62 +/- 11.41 pg/mL) and in stable asthmatics (64.22 +/- 10.31 pg/mL) compared to the control group subjects (50.40 +/- 30.70 pg/mL; p < 0.01). However, the difference was not statistically significant between the acute exacerbation and stable asthmatics groups (p > 0.05). Serum IL-8 is a poor indicator of disease activity in acute asthma; therefore, monitoring by serum IL-8 concentration is of limited value. The clinical value of serum IL-8 as a marker of disease activity remains to be established.

Topics: Acute Disease; Adolescent; Asthma; Biomarkers; Child; Child, Preschool; Eosinophils; Humans; Interleukin-8; Leukocyte Count; Prednisolone

Chemokines and cellular adhesion molecules are crucial determinants of the migration of immune effector cells to the tissues asthma and chronic obstructive pulmonary disease (COPD) are a complex of conditions, which have airflow limitation in common. The aim of this study was to determine the numbers and percentages of lymphocytes expressing adhesion molecules: LFA-1, ICAM-1 together with assessment of chemokines concentrations: IL-8 and MCP-1 in bronchoalveolar lavage fluid (BAL) of patients with asthma or chronic obstructive pulmonary disease (COPD). 12 patients with asthma, 14 patients with COPD, and 6 subjects of control group took part in this study. The expression of LFA-1 and ICAM-1 was assessed on lymphocytes by using immunohistochemistry (streptavidyn-biotin, DAKO, Denmark). ELISA test was used to measure IL-8 and MCP-1 concentrations in BAL (kits from R&D, USA). The percentage of lymphocytes expressing LFA-1 and ICAM-1 were: 33.9 +/- 23.8% and 25.8 +/- 12.2% in COPD patients, 23.9 +/- 12.1% and 15.3 +/- 4.42% in asthma patients, and 14.2 +/- 10% and 5.2 +/- 1.6% in the control group respectively. There was observed significant difference between the percentage of lymphocytes expressing LFA-1 and ICAM-1 of COPD and the control group. The concentrations of IL-8 were: 2306 +/- 1501 pg/ml in COPD, 233 +/- 27.3 pg/ml in asthma and 64 +/- 28.7 in the control group (p < 0.05). The concentrations of MCP-1 were: 768.9 +/- 668.1 pg/ml in COPD, 126.8 +/- 30.8 pg/ml in asthma, and 83.0 +/- 16.4 pg/ml in the control group (p < 0.05). There was observed correlation between lymphocytes expressing LFA-1 and IL-8 concentration (r = +0.5, p < 0.05) and between lymphocytes expressing LFA-1 and MCP-1 concentration (r = +0.5, p < 0.05), and between lymphocytes expressing ICAM-1 and MCP-1 concentration (r = +0.4, p < 0.05) only in COPD patients. Our data suggest that LFA-1 and ICAM-1 are important molecules in the recruitment of leukocytes and together with IL-8 and MCP-1 may have a role in pathomechanism of inflammation in asthma and especially in COPD.

Topics: Adult; Asthma; Bronchoalveolar Lavage Fluid; Cell Adhesion Molecules; Chemokine CCL2; Chemokines; Female; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Lung Diseases, Obstructive; Lymphocyte Function-Associated Antigen-1; Lymphocytes; Male

The aim of this study was to evaluate whether ozone exposure induces a similar airway inflammatory response in subjects with different degrees of asthma severity. Two groups of asthmatic subjects were studied: seven with intermittent mild asthma not requiring regular treatment (group A); and seven with persistent mild asthma requiring regular treatment with inhaled corticosteroids and long-acting beta2-agonists (group B). All subjects were exposed, in a randomized cross-over design, to air or O3 (0.26 parts per million (ppm) for 2 h with intermittent exercise); subjects in group B withdrew from regular treatment 72 h before each exposure. Before the exposure, and 1 and 2 h after the beginning of the exposure they performed a pulmonary function test, and a questionnaire was completed to obtain a total symptom score (TSS). Six hours after the end of the exposure, hypertonic saline (HS) sputum induction was conducted. Sputum cell percentages, eosinophil cationic protein (ECP) and interleukin (IL)-8 concentrations in the sputum supernatant were measured. TSS significantly increased and forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) significantly decreased after O3 exposure in comparison with air exposure in group A, whereas no changes were observed in group B except for a significant decrement of FEV1 2 h after the beginning of O3 exposure. Sputum neutrophil percentage was significantly higher after O3 exposure than after air exposure in both groups (Group A: 70.2% (28-87) versus 26.6% (8.6-73.2); Group B: 62.1% (25-82.4) versus 27.9% (14.4-54)). IL-8 was higher in sputum supernatant collected 6 h after O3 exposure than after air, only in group A. No change due to O3 has been found in sputum eosinophil percentage and ECP concentration in both groups. In conclusion, the degree of airway response to a short-term exposure to ozone is different in subjects with asthma of different severity. The available data do not allow elucidation of whether this difference depends on the severity of the disease or on the regular anti-inflammatory treatment.

Topics: Adolescent; Adult; Asthma; Blood Proteins; Cross-Over Studies; Eosinophil Granule Proteins; Eosinophils; Female; Forced Expiratory Volume; Humans; Inflammation; Inflammation Mediators; Interleukin-8; Leukocyte Count; Male; Neutrophils; Oxidants, Photochemical; Ozone; Peak Expiratory Flow Rate; Ribonucleases; Single-Blind Method; Sputum

Nitric oxide (NO) may have a role in the pathophysiology of tissue injury in response to inhaled ozone in animals.. A double blind, randomised, placebo controlled, crossover study was undertaken to investigate the effects of inhaled ozone in 10 normal and 10 atopic asthmatic volunteers. Subjects were exposed to 200 ppb ozone or clean air for four hours with intermittent exercise, followed by hourly measurement of spirometric parameters and exhaled NO for four hours. Nasal NO and methacholine reactivity were measured and exhaled breath condensate and induced sputum samples were collected four and 24 hours after exposure.. Exposure to ozone caused a fall in forced expiratory volume in one second (FEV(1)) of 7% in normal subjects (p<0.05) and 9% in asthmatic subjects (p<0.005). There was a 39% increase in sputum neutrophils at four hours in normal subjects (p<0.05) and a 35% increase at four hours in asthmatic subjects, remaining high at 24 hours (p<0.005 and p<0.05, respectively). There were no differences between normal and asthmatic subjects. There were no changes in methacholine reactivity, exhaled or nasal NO, nitrite levels in exhaled breath condensate, or sputum supernatant concentrations of interleukin 8, tumour necrosis factor alpha, or granulocyte-macrophage colony stimulating factor in either group.. Exposure to 200 ppb ozone leads to a neutrophil inflammatory response in normal and asthmatic subjects but no changes in exhaled NO or nitrite levels.

Topics: Adult; Asthma; Breath Tests; Cell Count; Cross-Over Studies; Double-Blind Method; Female; Forced Expiratory Volume; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Male; Nitric Oxide; Oxidants, Photochemical; Ozone; Sputum; Tumor Necrosis Factor-alpha; Vital Capacity

We investigated correlations between ozone-induced increases in inflammatory markers in induced sputum and in bronchial lavage fluid. Sixteen volunteers with intermittent asthma participated in a placebo-controlled parallel study with two exposures. Six days before and 16 h after the first exposure to ozone (0.4 ppm during 2 h) sputum was induced with hypertonic saline. This resulted in a significant increase in the sputum levels of eosinophil cationic protein (ECP; 1.8-fold; p = .03), neutrophil elastase (5.0-fold; p = .005) and the total cell number (1.6-fold; p = .02). After 4 weeks, a second exposure was randomized for air or ozone. Six days before and 16 h after the second exposure a bronchial lavage was performed. ECP values in sputum and in bronchial lavage fluid obtained after ozone correlated significantly (Rs = .79; p = .04), as did interleukin-8 (IL-8) values (Rs = .86; p = .01), and the percentage eosinophils (Rs = .89; p = .007). Moreover, the ozone-induced changes in percentage eosinophils observed in sputum and lavage fluid were highly correlated (Rs = .93; p = .003). In conclusion, changes in eosinophils, IL-8, and ECP markers induced by ozone and measured in sputum reflect the inflammatory responses in the lower airways of asthmatics, and may provide a noninvasive tool in epidemiologic studies on air pollution and asthma.

Topics: Adult; Asthma; Biomarkers; Blood Proteins; Bronchoalveolar Lavage Fluid; Bronchoconstrictor Agents; Eosinophil Granule Proteins; Eosinophils; Female; Humans; Inflammation; Interleukin-8; Leukocyte Count; Leukocyte Elastase; Male; Methacholine Chloride; Ozone; Placebos; Ribonucleases; Sputum

Inhalation of lipopolysaccharide (LPS) causes an inflammatory response in the lungs. To explore this response, inflammatory indices were measured in induced sputum from atopic asthmatic patients and compared with atopic and non-atopic subjects after inhalation of LPS.. The effects of inhaled LPS (60 micrograms) or placebo (0.9% saline) were examined in a randomised, double blind, crossover trial in 11 non-atopic normal subjects, seven atopic, non-asthmatic individuals, and eight atopic, asthmatic patients. Sputum was induced by inhalation of 3.5% saline before the test inhalation and again at six hours and 24 hours. Spirometry (forced expiratory volume in one second (FEV1), forced vital capacity (FVC)), heart rate, blood pressure, and temperature were recorded before challenge and at intervals until eight hours, and at 24 hours after challenge.. There was no change in cardiovascular parameters or spirometry with either exposure in any group. In the asthmatic patients only, inhalation of LPS caused a rise in temperature, with a peak of 0.6 degree C at seven hours, which was significantly higher than following placebo inhalation (p < 0.05). In normal subjects, LPS caused a significant rise in absolute neutrophil counts at 24 hours compared with placebo (median 1.1 x 10(6) cells/ml after LPS; median 0.2 x 10(6) cells/ml after placebo, p < 0.01), but no change in differential counts. In asthmatic patients, LPS caused a significant rise in differential neutrophil counts at six hours compared with placebo (median 88% after LPS; median 56% after placebo, p < 0.05), but no change in absolute cell counts at any time point. There was no change in neutrophil counts in the atopic subjects. There was a significant rise in sputum interleukin 8 (IL-8) concentrations in normal subjects at six hours compared with placebo (mean placebo 1.1 ng/ml; LPS 3.0 ng/ml, p < 0.05) and in asthmatics at 24 hours (mean placebo 2.0 ng/ml, LPS 6.9 ng/ml, p < 0.05). There were no changes in sputum concentrations of tumour necrosis factor alpha or granulocyte macrophage colony stimulating factor at any time.. Inhalation of LPS causes a neutrophilic inflammation with increases in IL-8 in both normal and asthmatic subjects.

Topics: Administration, Inhalation; Adult; Analysis of Variance; Asthma; Body Temperature; Cross-Over Studies; Double-Blind Method; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Hypersensitivity, Immediate; Interleukin-8; Leukocyte Count; Lipopolysaccharides; Lung; Neutrophils; Spirometry; Sputum; Tumor Necrosis Factor-alpha

Asthma exacerbations are closely associated with respiratory virus infections. However, the pathophysiological consequences of such infections in asthma are largely unclear.. To examine the effect of rhinovirus 16 (RV16) infection on airway hypersensitivity to histamine, and on interleukin-8 (IL-8) in nasal lavage.. Twenty-seven non-smoking atopic, mildly asthmatic subjects participated in a placebo-controlled, parallel study. A dose of 0.5-2.9 x 10(4) TCID50 RV16 or placebo was nasally administered. Cold symptoms were recorded by questionnaire throughout the study. Histamine challenges were performed at entry, and on days 4 and 11 after inoculation. Nasal lavages were obtained at entry, and on days 2 and 9. The response to histamine was measured by PC20 (changes expressed as doubling doses: DD) IL-8 levels were obtained by ELISA, and were expressed in ng/ml.. RV infection was confirmed by culture of nasal lavage and/or by antibody titre rise in each of the RV16-treated subjects. Among the 19 RV 16-treated subjects, eight developed severe cold symptoms. Baseline FEV1, did not change significantly during the study in either treatment group (P = 0.99). However, in the RV16-treated subjects there was a decrease in PC20 at day 4, which was most pronounced in those with a severe cold (mean change +/- SEM: -1.14 +/- 0.28 DD, P = 0.01). In addition, IL-8 levels increased in the RV16 group at days 2 and 9 (P < 0.001). The increase in nasal IL-8 at day 2 correlated significantly with the change in PC20 at day 4 (r = -0.48, P = 0.04).. We conclude that the severity of cold, as induced by experimental RV16 infection, is a determinant of the increase in airway hypersensitivity to histamine in patients with asthma. Our results suggest that this may be mediated by an inflammatory mechanism, involving the release of chemokines such as IL-8.

Topics: Adolescent; Adult; Asthma; Bronchial Hyperreactivity; Bronchial Provocation Tests; Common Cold; Double-Blind Method; Enzyme-Linked Immunosorbent Assay; Female; Forced Expiratory Volume; Histamine; Humans; Interleukin-8; Leukocyte Count; Lung; Male; Nasal Lavage Fluid; Rhinovirus

Airway inflammation is a very important feature of asthma and occurs simultaneously with increased hyperreactivity. We have examined whether the local inflammation provoked by histamine and allergen challenge of patients with atopic bronchial asthma is associated with the appearance, in vivo interleukin-B (IL-8) in bronchoalveolar lavage fluid (BAL). Bronchoscopy and BAL for further IL-8 investigation were performed before and 24 h after challenge test with histamine (n = 11), grass pollen antigen (n = 8) and PBS (n = 5). ELISA test was used to measure IL-8 concentration (pg/ml) (kits from R&D, USA). There was observed increased level of IL-8 (p < 0.05) after histamine and allergen challenge test. This increased level of IL-8 was correlated with neutrophils in BAL (Kendall's correlation coefficient = +0.5). We conclude that IL-8 may participate in creation of bronchial hyperreactivity in atopic bronchial asthma.

Topics: Adult; Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Female; Humans; Interleukin-8; Male; Respiratory Function Tests; Respiratory Hypersensitivity

Ozone may play a significant role in the exacerbation of airway disease in asthmatics, either by priming the airway mucosa such that cellular responses to allergen are enhanced or by exerting an intrinsic effect on airway inflammation. Previous investigations of nonasthmatic subjects revealed that ozone induces both nasal and bronchial inflammation, suggesting that nasal responses to ozone may be used as a surrogate marker for the effect of this pollutant on bronchial mucosal inflammation. In this study, the effect of exposure to 0.4 ppm ozone on nasal inflammation in 11 allergic asthmatics sensitive to Dermatophygoides farinae was examined. This study was designed such that the effect of ozone exposure on the late-phase reaction to allergen was emphasized, using eosinophil influx and changes in eosinophil cationic protein as principal endpoints. By employing a "split-nose" design, in which allergen was applied to only one side of the nose while saline was applied to the contralateral side, both the effect of ozone on nasal inflammation due to allergen challenge as well as its direct action on non-allergen-challenged nasal tissues was examined. The results reported herein indicate that ozone exposure has both a priming effect on allergen-induced responses as well as an intrinsic inflammatory action in the nasal airways of perennially allergic asthmatics.

Topics: Adolescent; Adult; Albumins; Allergens; alpha 1-Antitrypsin; Animals; Asthma; Bronchial Provocation Tests; Double-Blind Method; Eosinophils; Female; Humans; Inflammation; Interleukin-8; Leukocyte Count; Male; Mites; Nasal Lavage Fluid; Nasal Mucosa; Ozone; Rhinitis, Allergic, Perennial

Increasing evidence suggests that cytokines play a role in airway inflammation by attracting and activating inflammatory cells. This may lead to epithelial cell damage and airway hyperresponsiveness. Bronchial provocative concentration of histamine causing a 20% fall in forced expiratory volume in 1 second was measured in patients with mild asthma, and bronchial biopsy specimens were stained for granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-8, and activated eosinophils (EG2) in the bronchial epithelium. The effect of inhaled beclomethasone dipropionate was also assessed in a placebo-controlled double-blind manner. There was a correlation between GM-CSF expression and EG2-staining cells (r = 0.484 p < 0.05) in the epithelium. Provocative concentration of histamine causing a 20% fall in forced expiratory volume in 1 second was correlated with GM-CSF expression (r = -0.462, p < 0.05). Treatment with inhaled beclomethasone dipropionate 500 micrograms twice a day led to a significant decrease in both the expression of GM-CSF (p < 0.01) and IL-8 (p < 0.02) and the number of EG2-staining cells (p < 0.01) in the epithelium. The changes in GM-CSF (r = 0.798, p < 0.01) and IL-8 (r = 0.653, p < 0.02) expression were correlated with the changes in EG2-staining cells after treatment. These results suggest that GM-CSF may influence eosinophil activation in the epithelium in vivo and participate in the etiology of bronchial hyperresponsiveness in mild asthma. Also, beclomethasone dipropionate may inhibit eosinophil activation partly by downregulating the expression of GM-CSF and IL-8 in the bronchial epithelium.

Topics: Administration, Inhalation; Adolescent; Adult; Asthma; Beclomethasone; Bronchi; Double-Blind Method; Eosinophils; Epithelium; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Male; Middle Aged

The aim of this study was to assess the ability of nedocromil sodium (NS) to prevent the immediate asthmatic reaction and the increase in the serum level of heat stable neutrophil chemotactic activity (HS-NCA) induced by antigen inhalation. In a double-blind, cross-over study, 13 atopic subjects affected with seasonal asthma underwent a bronchial provocation test with a preselected dose of grass pollen allergen (enough to cause a decrease of > or = 20% in FEV1:FEV1 PD20) after pre-treatment with 4 mg NS or placebo. Serum samples were withdrawn from 11 subjects for HS-NCA determination. After NS administration the decrease in FEV1 was significantly less than after placebo administration at all time points after challenge (2 min P = 0.0004; 7 min, P = 0.0005; 17 min P = 0.0002 and 27 min P = 0.0005). The percentage increase in HS-NCA was significantly higher after placebo than after NS inhalation, both 10 (P = 0.0048) and 20 (P = 0.0068) min after challenge. Our study confirms previous investigations, showing that NS inhibits the immediate asthmatic response to allergen inhalation in atopic, asthmatic subjects and moreover it shows that this drug prevents in vivo the increase of the serum HS-NCA. This last finding has not been previously reported.

Topics: Adolescent; Adult; Asthma; Bronchial Provocation Tests; Bronchial Spasm; Chemotaxis, Leukocyte; Double-Blind Method; Female; Forced Expiratory Volume; Humans; Interleukin-8; Male; Middle Aged; Nedocromil; Neutrophils

Changes in chemical mediators after antigen challenge and exercise challenge tests were studied in children with asthma. Chemical mediators studied after antigen challenge and exercise challenge included histamine, leukotrienes (LTB4, LTC4, and LTD4), and neutrophil chemotactic factor of anaphylaxis (NCA). The pharmacologic modification of immediate and late-phase reactions was evaluated for procaterol (beta 2-agonist), cromolyn sodium, and prednisolone. Histamine levels were noted to rise in patients who had a dual response of a mild to moderate nature, but did not change in patients who had severe asthma. During exercise challenge cromolyn sodium inhibited both immediate and late-phase reactions but also inhibited plasma generation of NCA. Prednisolone, on the other hand, did not affect immediate reactions, but blocked late-phase reactions to exercise and also decreased NCA generation. Procaterol inhibited the generation of LTD4 and also inhibited NCA when compared with placebo. Exercise challenge did not alter levels of plasma histamine or of LTB4 or LTC4.

Topics: Adolescent; Animals; Asthma; Bronchial Provocation Tests; Chemotactic Factors; Child; Cromolyn Sodium; Double-Blind Method; Ethanolamines; Exercise Test; Histamine; Humans; Hypersensitivity, Delayed; Interleukin-8; Mites; Prednisolone; Procaterol; Respiratory Function Tests; SRS-A

The effect of pharmacologic agents on mast cell mediator release was investigated in vivo. Eight atopic asthmatic subjects with airways relatively unreactive to nonspecific stimuli (geometric mean PC20 methacholine, 4.0 mg/ml) underwent single-concentration allergen challenge before (control) or after inhaling albuterol 200 micrograms, cromolyn sodium 20 mg, or 0.9% sodium chloride placebo. Six of the same subjects also underwent allergen challenge after pretreatment with ipratropium bromide, 1 mg. Airway responses to pharmacologic agents and bronchial challenge were measured by change in both specific airway conductance (SGaw) and FEV1. Mast cell mediator release was monitored by serial change in plasma histamine and, in addition, serum neutrophil chemotactic factor (NCF) on the placebo, albuterol, and cromolyn sodium challenge days. Control and placebo allergen challenges were associated with repeatable mean maximal falls in SGaw (48.5 versus 49.6%) and FEV1 (25.7 versus 25.5%). The mean increments in plasma histamine were not significantly different on the control (0.17 to 0.44 ng/ml) or placebo challenge days (0.18 to 0.64 ng/ml), with maximal levels occurring 5 min after challenge. A sustained increase in NCF was identified on the placebo challenge day (155.0% above baseline). Pretreatment with albuterol abolished any significant bronchoconstriction, with mean maximal falls in SGaw and FEV1 after challenge of 7.5 and 1.4%, respectively. These changes in airway caliber were not associated with any significant increment in mean plasma histamine (0.17 to 0.22 ng/ml) or serum NCF (4.1% increase). Cromolyn sodium pretreatment, while attenuating the airway response, was still associated with significant falls in SGaw (22.7%) and FEV1 (7.3%) and increases in plasma histamine (0.18 to 0.27 ng/ml).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Albuterol; Asthma; Atropine Derivatives; Bronchial Provocation Tests; Chemotactic Factors; Cromolyn Sodium; Female; Forced Expiratory Volume; Histamine; Humans; Interleukin-8; Ipratropium; Male; Middle Aged; Respiratory System

Topics: Administration, Intranasal; Airway Obstruction; Animals; Asthma; Bronchial Spasm; Chemotactic Factors; Clinical Trials as Topic; Cromolyn Sodium; Double-Blind Method; Guinea Pigs; Haplorhini; Histamine Release; Humans; Interleukin-8; Long-Term Care; Mast Cells; Sulfur Dioxide; Theophylline

Epithelial cytokines are involved in the orchestration of T1/T2 inflammatory patterns. We question the persistence of this trait in air-liquid interface (ALI) epithelial cultures and whether this local orientation can be related to systemic patterns (e.g., blood eosinophil counts [BECs]). We investigated alarmin release related to high versus low T2 phenotypes associated with chronic airway diseases. ALIs were reconstituted from 32 control, 40 chronic obstructive pulmonary disease, and 20 asthmatic patients. Interleukin-8 (IL-8; a T1-cytokine), IL-25, IL-33, and thymic stromal lymphopoietin (T2-alarmins) concentrations were assessed in subnatants at steady state and used to explain blood neutrophil and eosinophil counts. IL-25 and IL-8 levels were highest in asthma ALI-subnatants, whereas IL-33 was sparsely detected. Thymic stromal lymphopoietin levels were similar among groups. All asthma cell cultures were T1-high/T2-high, while chronic obstructive pulmonary disease and controls tended to be mixed. BECs were independently explained by both disease and in-culture T2-alarmin levels, irrespective of the T2-alarmin considered. The epithelial ALI-T2 signature was more frequently high in patients with a BEC > 300/mm

Topics: Alarmins; Asthma; Cytokines; Eosinophils; Humans; Interleukin-33; Interleukin-8; Pulmonary Disease, Chronic Obstructive; Thymic Stromal Lymphopoietin

Toluene diisocyanate (TDI)-induced asthma is characterized by mixed inflammation dominated by neutrophils, and is refractory to steroid treatment. Neutrophil extracellular traps (NETs) play an important role in severe asthma, but their role in TDI-induced asthma models is unclear. This study focused on the role and mechanism of NETs in steroid-resistant TDI-induced asthma.. Induced sputum was collected from 85 asthmatic patients and 25 healthy controls to detect eDNA. A murine TDI-induced asthma model was prepared, and asthmatic mice were given dexamethasone or DNase I. In vitro, the human bronchial epithelial cell line HBE was stimulated with NETs or TDI-human serum albumin (TDI-HSA).. Asthma patients had higher sputum eDNA compared to healthy subjects. In asthma patients, eDNA was positively correlated with sputum neutrophils, and negatively correlated with FEV1%predicted. Airway inflammation, airway reactivity, Th2 cytokine levels in lymph supernatant, and levels of NETs were significantly increased in the TDI-induced asthmatic mice. These increases were suppressed by DNase I, but not by dexamethasone. Inhibition of NETs improved interleukin (IL)-8 and MKP1 mRNA expression, and reduced phosphorylation of GR-S226 induced by TDI. Inhibition of NETs improved airway epithelial barrier disruption, as well as p38 and ERK signaling pathways in TDI-induced asthmatic mice. In vitro, NETs promoted the expression of IL-8 mRNA in HBE cells, and reduced the expression of MKP1. IL-8 elevation induced by NETs was suppressed by a p38 inhibitor or ERK inhibitor, but not by dexamethasone. Pretreatment with RAGE inhibitor reduced NETs induced p38/ERK phosphorylation and IL-8 levels in HBE cells.. Our data suggest that targeting NETs might effectively improved TDI-induced airway inflammation and airway epithelial barrier function. This may potentially be a treatment for patients with steroid-resistance asthma.

Topics: Animals; Asthma; Dexamethasone; Disease Models, Animal; Extracellular Traps; Humans; Inflammation; Interleukin-8; Mice; Steroids; Toluene 2,4-Diisocyanate

Heat shock protein 70 (HSP70) regulates the ligand binding of the glucocorticoid receptor (GR). In asthma patients, heat treatment increased both the GR expression and secretion of extracellular HSP70 (eHSP70) by bronchial epithelial cells (EC). The objective of this study was to assess the effects of eHSP70 on GR expression and the GR-dependent regulation of immune response in human bronchial ECs. Cells were treated with either eHSP70 or transfected with an expression vector for intracellular HSP70 (iHSP70). Ribonucleic acid (RNA) and protein levels were detected by reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, and immunofluorescence. Interleukin (IL-6 and IL-8) secretion was determined by enzyme linked immunosorbent assay (ELISA). The overexpression of iHSP70 decreased, while eHSP70 increased GR expression. In addition, eHSP70 increased the expression of the GR target dual-specificity phosphatase 1 (DUSP-1). In doing so, eHSP70 reduced the tumor growth factor (TGF)-β1-dependent activation of extracellular signal-regulated kinase (Erk)-1/2 and cyclic AMP response element binding protein (CREB) and the secretion of IL-6 and IL-8. Blocking the GR or Toll-like receptor 4 (TLR4) counteracted all eHSP70-induced effects. This study demonstrates a novel anti-inflammatory effect of eHSP70 by the signaling cascade of TLR4-GR-DUSP1, which inhibits TGF-β1-activated pro-inflammatory ERK1/2-CREB signaling and cytokine secretion. The findings suggest that eHSP70 might present a novel non-steroidal therapeutic strategy to control airway inflammation in asthma.

Topics: Asthma; Dual-Specificity Phosphatases; Epithelial Cells; HSP70 Heat-Shock Proteins; Humans; Inflammation; Interleukin-6; Interleukin-8; Receptors, Glucocorticoid; Toll-Like Receptor 4

Equine asthma is an airway disease that affects a large number of horses annually leading to considerable economic losses in the horse industry. Despite advances in research in this area, there is still a lack of information on its etiology and molecular characterization in pasture associated asthma. The objective of the current study was to characterize the inflammatory disease of lower airways in horses maintained on pasture through cytologic and immunologic profile during the summer in a tropical environment by analysis of the gene expression of Th1 cytokines (IFN- λ, IL-8), Th2 cytokines (IL-4 and IL-5), and pro-inflammatory cytokines (IL-1, TNF-α) in the bronchoalveolar lavage (BAL) fluid in healthy and asthma horses on pasture. A group 39 of clinically healthy horses maintained on native pasture and supplemented with concentrate was evaluated by BAL analyzed for differential cellular count and assigned into a control and an asthma group. The gene expression of pro-inflammatory cytokines was analyzed in the BAL by reverse time PCR (RT-PCR) (IL-1α (alpha), IL-4, IL-5, IL-8, TNF-α alpha and IFN-λ), using β-actin as housekeeping gene. Higher gene expression of IL-1, IL-4, IL-5, IL-8, IFN-λ in the BAL of asthma horses was found. Current results indicate an increase in Th2, characterizing an allergic inflammatory reaction due to the significant increase in IL-5 in asthmatic horses (10.3 ± 1.13), when compared to the values ​​obtained in normal horses (3.27 ± 0.46). The only down regulated cytokine in the asthma group was TNF-α, suggesting a chronic antigenic reaction.

Topics: Animals; Asthma; Horse Diseases; Horses; Interleukin-4; Interleukin-5; Interleukin-8; Tumor Necrosis Factor-alpha

Secretion of translationally controlled tumor protein (TCTP) was found in body fluids during the late phase of allergic reactions, implicating TCTP in allergic diseases. Furthermore, blocking TCTP has been shown to be helpful in treating asthma and allergies in animal models. The objectives of this study were to produce anti-TCTP monoclonal antibodies (mAbs), test their ability to inhibit the cytokine-like function of dimeric TCTP (dTCTP) in vitro and to assess their therapeutic effects in a murine model of ovalbumin (OVA)-induced airway inflammation. We first verified the inhibitory effects of 4 anti-TCTP mAbs on dTCTP-induced secretion of IL-8 in BEAS-2B cells. To investigate the anti-inflammatory effect of anti-TCTP mAbs on allergic airway inflammation, we treated OVA-sensitized mice with anti-TCTP mAbs before OVA challenge. The changes in bronchoalveolar lavage fluid (BALF) cells, IL-4, IL-5, and IL-13 levels in both BALF and lung homogenates, plasma levels of OVA-specific IgE, and lung tissues were analyzed. We found that JEW-M449 anti-TCTP mAb bound to the flexible loop of TCTP and significantly inhibited dTCTP-induced IL-8 release, making it the most effective inhibitor in our study. We also found that treatment with JEW-M449 significantly reduced the infiltration of inflammatory cells and suppressed the OVA-induced upregulation of type 2 cytokines in both BALF and lung homogenates in a dose-dependent manner. In addition, JEW-M449 significantly attenuated the degree of goblet cell hyperplasia and mucus secretion. Our results demonstrate that specific targeting of the flexible loop of TCTP is a potent strategy for treating airway inflammatory diseases.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hypersensitivity; Inflammation; Interleukin-8; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Tumor Protein, Translationally-Controlled 1

Asthma progression is involved in airway epithelial dysfunction, airway inflammatory response, and mucus hypersecretion. Euxanthone has been found to exhibit cytotoxic activity on several human diseases, such as neurological disorders and cancers. Our study aimed to explore the influence of euxanthone on lipopolysaccharide (LPS)-induced injury, inflammatory response, and mucin 5AC (MUC5AC) hypersecretion in human airway epithelial cells (AECs). Network pharmacology analysis was carried out to analyze the drug targets and key pathways of euxanthone against asthma. Cell injury was evaluated by CCK-8, Lactate dehydrogenase (LDH) release assay, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. The production of interleukin (IL)-6, IL-8, monocyte chemoattractant protein-1 (MCP-1), and MUC5AC was measured using enzyme-linked immunosorbent assay (ELISA). MUC5AC mRNA expression was detected by qRT-PCR. Toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88) protein expression was examined by western blot analysis. Venn diagram showed 14 overlapping targets between euxanthone and asthma. According to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, we focused on TLR signaling pathway. LPS exposure evoked viability reduction, increased LDH release and apoptosis, and induced production of inflammatory cytokines (IL-6, IL-8, and MCP-1) and MUC5AC hypersecretion in human AECs, which were alleviated by euxanthone. Mechanistically, we validated that euxanthone attenuated LPS-induced activation of TLR4/MyD88 pathway in AECs. Moreover, inhibition of the TLR4/MyD88 pathway enhanced the inhibitory effect of euxanthone on LPS-induced cell injury, inflammatory response and MUC5AC expression. In conclusion, euxanthone attenuated LPS-induced cell injury, inflammatory response, and MUC5AC expression in AECs by inhibiting the activation of TLR4/MyD88 pathway.

Topics: Asthma; Epithelial Cells; Humans; Interleukin-8; Lipopolysaccharides; Mucin 5AC; Myeloid Differentiation Factor 88; Toll-Like Receptor 4; Xanthones

Pediatric bronchial asthma signifies a frequent chronic inflammatory airway disorder influencing many children. Despite its irrefutable importance, its exact pathogenesis is not completely elucidated.. The study aimed to investigate the correlation between mitophagy machinery proteins, ER stress biomarkers and total polyamine and their role in disease progression via targeting NF-κB mechanisms.. Sixty children with atopic bronchial asthma were enrolled in the study, they were allocated into 2 equal groups (mild/moderate and severe atopic asthmatic groups). Thirty age-matched healthy control subjects were also included in the study to represent the control group. Phosphatase and tensin homolog (PTEN)-induced kinase-1 (PINK-1) and Parkin messenger RNA (mRNA) expressions were assessed by (RT-PCR) technique. Levels of inositol requiring enzyme 1α (IRE1α), total polyamines, interleukin 6 & 8 (IL-6, IL-8) and nuclear factor kappa B (NF-κB) were assessed by enzyme-linked immunosorbent assay. Oxidative stress (OS) biomarkers were also measured.. PINK-1 and PARK mRNA expressions were significantly upregulated in asthmatic patients. Likewise, the level of IRE1α, total polyamines, inflammatory cytokines, and OS biomarkers were significantly elevated in asthmatic groups comparing to control group with the highest levels noticed in severe atopic asthmatic group.. the study documented a correlation between mitophagy machinery proteins, ER stress biomarkers and total polyamines that may pave a new platform to understand pediatric asthma pathogenesis and could be used as reliable biomarkers to evaluate disease progression.

Topics: Asthma; Case-Control Studies; Child; Disease Progression; Endoplasmic Reticulum Stress; Endoribonucleases; Female; Humans; Interleukin-6; Interleukin-8; Male; Mitophagy; NF-kappa B; Polyamines; Protein Kinases; Protein Serine-Threonine Kinases; Signal Transduction; Ubiquitin-Protein Ligases; Up-Regulation

Neutrophilic asthma (NA) is a severe type of steroid resistant asthma, and so far the immune mechanisms underlying NA are not clear. In this article, we performed a comprehensive assessment of Th-cell subsets and cytokines in severe NA patients. A total of 13 healthy individuals and 31 severe asthma patients were enrolled in this study. Refractory asthma patients were defined as those with eosinophilic asthma (EA, accounted for 32% of asthmatic patients) or NA (68%) according to sputum neutrophil/eosinophil counts or blood eosinophils. Th-cell subsets in peripheral blood mononuclear cells (PBMCs) were analyzed by flow cytometry, and cytokines were detected by cytometric bead array (CBA). The results showed significant differences were observed in Th-cell phenotypes, where the number of Th1 cells were reduced and the numbers of Th2 cells were increased in NA and EA groups, respectively, when compared with healthy controls. Th17 cells were not strongly associated with severe neutrophilic asthma. The frequencies of mucosal-associated invariant T (MAIT) cells were strikingly reduced in severe asthma patients, especially in the NA group. This NA group also showed increased levels of IL-17A, IL-17F, TNF-α, and IL-6 in serum and increased levels of IL-17A, IL-17F, IFN-γ, TNF-α, IL-1β, IL-5, IL-6, and IL-8 in sputum. In addition, sputum IL-6 was positively correlated with TNF-α, IFN-γ, IL-17A, and IL-8. Our results uncovered a controversial role for Th17 cells, which were reduced in severe asthma patients. Severe neutrophilic asthma was associated with a striking deficiency of MAIT cells and high pro-inflammatory cytokine levels.

Topics: Asthma; Cytokines; Humans; Interleukin-17; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Th17 Cells; Tumor Necrosis Factor-alpha

Asthma is characterised by an aggravated immune response to respiratory viral infections. This phenomenon is a clinically well-recognised driver of acute exacerbations, but how different phenotypes of asthma respond immunologically to viruses is unclear.. To describe the association between different phenotypes and severity of asthma and bronchial epithelial immune responses to viral stimulation.. In the Immunoreact study, healthy subjects (n=10) and 50 patients with asthma were included; 30 (60%) were atopic, and 34 (68%) were eosinophilic; 14 (28%) had severe asthma. All participants underwent bronchoscopy with collection of bronchial brushings. Bronchial epithelial cells (BECs) were expanded and stimulated with the viral replication mimic poly (I:C) (Toll-like receptor (TLR)3 agonist). Patients with atopic asthma had increased induction of interleukin (IL)-4, interferon (IFN)-β, IL-6, tumour necrosis factor-α, and IL-1β after poly (I:C) stimulation compared to non-atopic patients, whereas in patients with eosinophilic asthma only IL-6 and IL-8 induction was higher than in non-eosinophilic asthma. Patients with severe asthma displayed a decreased antiviral IFN-β, and increased expression of IL-8, most pronounced in atopic and eosinophilic asthmatics. Furthermore, induction of IL-33 in response to poly (I:C) was increased in severe atopic and in severe eosinophilic asthma, but thymic stromal lymphopoietin only in severe eosinophilic asthma.. The bronchial epithelial immune response to a viral mimic stimulation differs between asthma phenotypes and severities, which may be important to consider when targeting novel asthma treatments.

Topics: Antiviral Agents; Asthma; Humans; Immunity; Interferon-beta; Interleukin-6; Interleukin-8; Phenotype; Poly I-C

Allergen exposure worsens viral-triggered asthma exacerbations and could predispose the host to secondary bacterial infections. We have previously demonstrated that exposure to house dust mite (HDM) reduced TLR-3-induced IFN-β in human bronchial epithelial cells (HBECs) from healthy donors. We hypothesize that HDM sensitization in different ways may be involved in both viral and bacterial resistance of HBECs in asthma. In this study, the role of HDM sensitization and effects of HDM exposure on viral stimulus-challenged HBECs from asthmatic donors have been explored with regard to expression and release of molecules involved in anti-viral and anti-bacterial responses, respectively.. HBECs from HDM-sensitized (HDM+) and unsensitized (HDM-) patients with asthma were used. HBECs were exposed to HDM or heat inactivated (hi)-HDM (20 μg/ml) for 24 h prior to stimulation with the viral infection mimic, Poly(I:C), for 3 or 24 h. Samples were analyzed with ELISA and RT-qPCR for β-defensin-2, IFN-β, TSLP, and neutrophil-recruiting mediators: IL-8 and TNF-⍺. NFκB signaling proteins p105, p65, and IκB-⍺ were analyzed by Western blot.. Poly(I:C)-induced IFN-β expression was reduced in HBECs from HDM + compared to HDM- patients (p = 0.05). In vitro exposure of HBECs to HDM furthermore reduced anti-microbial responses to Poly(I:C) including β-defensin-2, IL-8, and TNF-⍺, along with reduced NFκB activity. This was observed in HBECs from asthma patients sensitized to HDM, as well as in non-sensitized patients. By contrast, Poly (I:C)-induced release of TSLP, a driver of T2 inflammation, was not reduced with exposure to HDM.. Using HBECs challenged with viral infection mimic, Poly(I:C), we demonstrated that allergic sensitization to HDM was associated with impaired anti-viral immunity and that HDM exposure reduced anti-viral and anti-bacterial defense molecules, but not TSLP, across non-allergic as well as allergic asthma. These data suggest a role of HDM in the pathogenesis of asthma exacerbations evoked by viral infections including sequential viral-bacterial and viral-viral infections.

Topics: Animals; Asthma; beta-Defensins; Dermatophagoides pteronyssinus; Humans; Interleukin-8; Poly I-C; Pyroglyphidae; Virus Diseases

Childhood asthma is a common chronic inflammatory lung disease in children, among which airway inflammation is the main driving factor of asthma symptoms. Follistatin-like protein 1 (FSTL1) is involved in multiple inflammatory processes, but its role in airway inflammation has not been fully elucidated.. We used lipopolysaccharide (LPS) to stimulate human primary bronchial epithelial (BEAS-2B) cells to establish an in vitro airway inflammation model. The expression of FSTL1 was detected by qPCR. Cell Counting Kit-8 and Annexin V-PI double staining was used to analyze the viability and apoptosis of BEAS-2B. The content of IL-6, IL-8 and TNF-α was determined by ELISA kit. Western blot was used to detect the protein expression level of the bone morphogenetic protein 4 (BMP4) and KLF4. The levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and malondialdehyde were measured to assess oxidative stress.. The mRNA expression of FSTL1 was significantly increased in LPS-treated BEAS-2B cells. Silencing of FSTL1 inhibited the release of IL-6, IL-8, TNF-α, and cell apoptosis as well as enhanced the activities of SOD, CAT, and GSH-Px. Silencing of FSTL1 reversed the inflammatory state of cells by upregulating BMP4 and increasing the expression level of KLF4.. Silencing of FSTL1 reduced LPS-induced BEAS-2B cell damage by regulating the BMP4/KLF4 axis. FSTL1 may be a potential target for the treatment of asthma.

Topics: Asthma; Bone Morphogenetic Protein 4; Child; Epithelial Cells; Follistatin-Related Proteins; Gene Silencing; Humans; Inflammation; Interleukin-6; Interleukin-8; Kruppel-Like Factor 4; Lipopolysaccharides; Oxidative Stress; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Asthma is a complex multifactorial chronic airway inflammatory disease with diverse phenotypes and levels of severity and is associated with significant health and economic burden. In a certain population of asthma patients, the symptoms cannot be well controlled with steroid. There has been long standing interest in the use of probiotics for treating allergic diseases. The purpose of this study is to investigate whether the combination of Lactobacillus rhamnosus GG (LGG) with prednisolone could reduce the dosage of glucocorticoid in controlling airway inflammation in a murine model for allergic asthma.. We used Der p 2-sensitized asthma model in female BALB/c mice. The animals were treated with 75 μl or 50 μl oral prednisolone or combination treatment of these two doses of oral prednisolone with LGG. Airway hyperresponsiveness, serum specific IgE/IgG1/IgG2a, infiltrating inflammatory cells in lung and cytokines were assessed.. Compared to 75 μl prednisolone, a lower dose of prednisolone with 50 μl was less satisfactory in suppressing airway hyperresponsives, serum IgE and IgG1, Th2 cytokines and inflammatory cytokines such as IL-6, IL-8 and IL-17 as well as infiltrating inflammatory cells. However, combination of 50 μl prednisolone and LGG decreased airway resistance and serum IgE and IgG1, inhibited the production of IL-4, IL-5, IL-6, IL-8, IL-13 and IL-17, upregulated serum IgG2a and enhanced Th1 immune response.. LGG may reduce the dosage of prednisolone and thus may be beneficial in the treatment of asthma.

Topics: Adrenal Cortex Hormones; Animals; Asthma; Cytokines; Disease Models, Animal; Female; Humans; Immunoglobulin E; Immunoglobulin G; Inflammation; Interleukin-17; Interleukin-6; Interleukin-8; Lacticaseibacillus rhamnosus; Mice; Mice, Inbred BALB C; Ovalbumin; Prednisolone

Nontypeable Haemophilus influenzae (NTHi) is a respiratory tract pathobiont that chronically colonizes the airways of asthma patients and is associated with severe, neutrophilic disease phenotypes. The mechanism of NTHi airway persistence is not well understood, but accumulating evidence suggests NTHi can persist within host airway immune cells such as macrophages. We hypothesized that NTHi infection of pulmonary macrophages drives neutrophilic inflammation in severe asthma.. Bronchoalveolar lavage (BAL) samples from 25 severe asthma patients were assessed by fluorescence in situ hybridisation to quantify NTHi presence. Weighted gene correlation network analysis (WGCNA) was performed on RNASeq data from NTHi-infected monocyte-derived macrophages to identify transcriptomic networks associated with NTHi infection.. NTHi was detected in 56% of BAL samples (NTHi+) and was associated with longer asthma duration (34 vs 22.5 years, p = .0436) and higher sputum neutrophil proportion (67% vs 25%, p = .0462). WGCNA identified a transcriptomic network of immune-related macrophage genes significantly associated with NTHi infection, including upregulation of T17 inflammatory mediators and neutrophil chemoattractants IL1B, IL8, IL23 and CCL20 (all p < .05). Macrophage network genes SGPP2 (p = .0221), IL1B (p = .0014) and GBP1 (p = .0477) were more highly expressed in NTHi+ BAL and moderately correlated with asthma duration (IL1B; rho = 0.41, p = .041) and lower prebronchodilator FEV1/FVC% (GBP1; rho = -0.43, p = .046 and IL1B; rho = -0.42, p = .055).. NTHi persistence with pulmonary macrophages may contribute to chronic airway inflammation and T17 responses in severe asthma, which can lead to decreased lung function and reduced steroid responsiveness. Identifying therapeutic strategies to reduce the burden of NTHi in asthma could improve patient outcomes.

Topics: Asthma; Haemophilus Infections; Haemophilus influenzae; Humans; Inflammation; Interleukin-8; Macrophages, Alveolar

Infection with rhinovirus (RV) is a major risk factor for disease exacerbations in patients with allergic asthma. This study analysed a broad set of cytokines in the noses of children and adults with asthma during RV infection in order to identify immunophenotypes that may link to virus-induced episodes.. Nasal wash specimens were analysed in children (n = 279 [healthy, n = 125; stable asthma, n = 64; wheeze, n = 90], ages 2-12) who presented to a hospital emergency department, and in adults (n = 44 [healthy, n = 13; asthma, n = 31], ages 18-38) who were experimentally infected with RV, including a subset who received anti-IgE. Cytokines were measured by multiplex bead assay and data analysed by univariate and multivariate methods to test relationships to viral load, allergic status, airway inflammation, and clinical outcomes.. Analysis of a core set of 7 cytokines (IL-6, CXCL8/IL-8, IL-15, EGF, G-CSF, CXCL10/IP-10 and CCL22/MDC) revealed higher levels in children with acute wheeze versus those with stable asthma or controls. Multivariate analysis identified two clusters that were enriched for acutely wheezing children; one displaying high viral load ("RV-high") with robust secretion of CXCL10, and the other displaying high IgE with elevated EGF, CXCL8 and both eosinophil- and neutrophil-derived mediators. Broader assessment of 39 cytokines confirmed that children with acute wheeze were not deficient in type 1 anti-viral responses. Analysis of 18 nasal cytokines in adults with asthma who received RV challenge identified two clusters; one that was "RV-high" and linked to robust induction of anti-viral cytokines and anti-IgE; and the other associated with more severe symptoms and a higher inflammatory state featuring eosinophil and neutrophil factors.. The results confirm the presence of different immunophenotypes linked to parameters of airway disease in both children and adults with asthma who are infected with RV. Such discrepancies may reflect the ability to regulate anti-viral responses.

Topics: Adolescent; Adult; Asthma; Chemokine CXCL10; Child; Child, Preschool; Cluster Analysis; Cytokines; Enterovirus Infections; Epidermal Growth Factor; Granulocyte Colony-Stimulating Factor; Humans; Interleukin-15; Interleukin-6; Interleukin-8; Picornaviridae Infections; Respiratory Sounds; Rhinovirus; Young Adult

We are currently screening human volunteers to determine their sputum polymorphonuclear neutrophil (PMN) response 6- and 24-hours following initiation of exposure to wood smoke particles (WSP). Inflammatory responders (≥10% increase in %PMN) are identified for their subsequent participation in mitigation studies against WSP-induced airways inflammation. In this report we compared responder status (<i>N</i> = 52) at both 6 and 24 hr time points to refine/expand its classification, assessed the impact of the GSTM1 genotype, asthma status and sex on responder status, and explored whether sputum soluble phase markers of inflammation correlate with PMN responsiveness to WSP.. Six-hour responders tended to be 24-hour responders and vice versa, but 24-hour responders also had significantly increased IL-1beta, IL-6, IL-8 at 24 hours post WSP exposure. The GSTM1 null genotype significantly (<i>p</i> &lt; 0.05) enhanced the %PMN response by 24% in the 24-hour responders and not at all in the 6 hours responders. Asthma status enhanced the 24 hour %PMN response in the 6- and 24-hour responders. In the entire cohort (not stratified by responder status), we found a significant, but very small decrease in FVC and systolic blood pressure immediately following WSP exposure and sputum %PMNs were significantly increased and associated with sputum inflammatory markers (IL-1beta, IL-6, IL-8, and PMN/mg) at 24 but not 6 hours post exposure. Blood endpoints in the entire cohort showed a significant increase in %PMN and PMN/mg at 6 but not 24 hours. Sex had no effect on %PMN response.. The 24-hour time point was more informative than the 6-hour time point in optimally and expansively defining airway inflammatory responsiveness to WSP exposure. GSTM1 and asthma status are significant effect modifiers of this response. These study design and subject parameters should be considered before enrolling volunteers for proof-of-concept WSP mitigation studies.

Topics: Asthma; Biomarkers; Genotype; Glutathione Transferase; Humans; Inflammation; Interleukin-6; Interleukin-8; Neutrophils; Smoke; Wood

Most of the asthma with low Th2 is severe steroid-resistant asthma, the exact pathogenesis of which has not yet been fully elucidated. We found that IL-6 and IL-8 were highly expressed in the sputum supernatant of severe asthma and ephrin type-A receptor 2 (EphA2) was highly expressed on bronchial epithelial cells. So, is there a connection between these two phenomena? To clarify this issue, we stimulated bronchial epithelial cells 16HBE with Dermatophagoides pteronyssinus and its compontents LPS, respectively, and detected the activation of EphA2, activation of downstream pathways and secretion of inflammatory cytokines. A mouse asthma model was established, and the therapeutic effects of inhibiting or blocking EphA2 on mouse asthma were investigated. The results showed that D. pteronyssinus and its component LPS phosphorylated EphA2 on 16HBE, activated downstream signaling pathways STAT3 and p38 MAPK, and promoted the secretion of IL-6 and IL-8. After knockout of EphA2 on 16HBE, the activation of inflammatory pathways was attenuated and the secretion of IL-6 and IL-8 was significantly reduced. Inhibition or blockade of EphA2 on mouse airways resulted in a significant reduction in airway hyperresponsiveness and airway inflammation, and a significant decrease in the expression levels of IL-6, IL-17F, IL-1α, IL-1β and TNF in bronchoalveolar lavage fluid and lung tissue. Our study uncovers a novel role for EphA2 expressed on airway epithelial cells in the pathogenesis of asthma; EphA2 recognizes D. pteronyssinus or its component LPS and promotes the secretion of IL-6 and IL-8 by airway epithelial cell, thereby mediating airway inflammation. Thus, it is possible to provide a new molecular therapy for severe asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Dermatophagoides pteronyssinus; Disease Models, Animal; Inflammation; Interleukin-6; Interleukin-8; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Receptor, EphA2

Asthma is a high-incidence disease in the world. Oxysophocarpine (OSC), a quinolizidine alkaloid displays various pharmacological functions including anti-inflammation, neuroprotective, anti-virus and antioxidant. Here, we established mice and cell asthmatic model to explore the effects of OSC for asthma treatment. Mice were sensitized and challenged with ovalbumin (OVA) and treated with OSC before challenge. Enzyme-linked immuno sorbent assay (ELISA), hematoxylin and eosin (H&E), periodic acid-schiff (PAS), tolonium chloride staining and immunohistochemical assay were performed. OSC treatment inhibited inflammatory cell infiltration and mucus secretion in the airway, reduced IgE level in mouse serum and decreased IL-4, IL-5 production in bronchoalveolar lavage fluid (BALF). OSC also reduced the spleen index to regulate immune function. Meanwhile, NCI-H292 cells were induced by lipopolysaccharide (LPS) to simulate airway epithelial injury. OSC pretreatment decreased the IL-6 and IL-8 cytokine levels, mucin 5 AC expression, and mucin 5 AC mRNA level in the cell model. Further, OSC suppressed the phosphorylation of c-Jun N-terminal kinase (JNK), and activator protein 1 (AP-1, Fos and Jun). These findings revealed that OSC alleviated bronchial asthma associated with JNK/AP-1 signaling pathway.

Topics: Alkaloids; Animals; Antioxidants; Asthma; Cytokines; Disease Models, Animal; Eosine Yellowish-(YS); Hematoxylin; Immunoglobulin E; Interleukin-4; Interleukin-5; Interleukin-6; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Molecular Structure; Mucins; Mucus; Ovalbumin; Periodic Acid; Quinolizidines; RNA, Messenger; Tolonium Chloride; Transcription Factor AP-1

Bronchial asthma is a chronic inflammatory airway illness. For the first time, we evaluated the proposed anti-asthmatic protective and therapeutic potency of inhaling Punica granatum juice (PJE) and peel (PPE) extract mixture (PM). Rats were challenged with ovalbumin (OVA) for 23 days and aerosolized with PM before each OVA challenge (protected group) or following the final OVA challenge for 3 days (therapeutic group). Considerable concentrations of phenolics were detected in PJE and PPE. Therefore, PM demonstrated synergistic scavenging abilities of NO and DPPH radicals. It also showed synergistic anti-inflammatory activities against lipopolysaccharide (LPS)-induced inflammation in the white blood cells by lowering the gene expression of CXCR1, CXCR2, IL-6, and IL-8. In addition, PM increased IL-10 gene expression while decreasing NO and TNF-α levels in LPS-exposed cells. Regarding the rats that were protected with PM, they exerted pulmonary pro-oxidant effects but prevented the OVA-induced upregulation of NF-κB, IKK, TNF-α, COX-2, iNOS, IL-13, and COL1A1, as well as MUC5AC and mucin over-secretion. While PM in the therapeutic group improved reactive oxygen species levels and normalized most of the investigated inflammatory and fibrotic mediators and mucin formation, but slightly improved the antioxidant indices. In addition, OVA-induced morphological alterations were massively improved after PM inhalation for short or long periods. Thus, PM inhalation prevented and treated OVA-induced pulmonary inflammation and fibrosis, while the inhalation period between 3 and 23 days needs to be optimized to acquire a better impact on the antioxidant indices.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Antioxidants; Asthma; Bronchoalveolar Lavage Fluid; Cyclooxygenase 2; Disease Models, Animal; Interleukin-10; Interleukin-13; Interleukin-6; Interleukin-8; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Mucins; NF-kappa B; Ovalbumin; Pomegranate; Rats; Reactive Oxygen Species; Respiratory Aerosols and Droplets; Signal Transduction; Tumor Necrosis Factor-alpha

Endogenous adenosine 5'-monophosphate (AMP), acetylcholine (ACh), and histamine (HA) are known to be important in bronchial contraction, but their clinical relevance to asthma is poorly understood. We aimed to quantify endogenous AMP, ACh, and HA in induced sputum samples and explore their relationships with asthma control and exacerbations.. 20 healthy subjects and 112 asthmatics underwent clinical assessment, sputum induction, and blood sampling. The level of asthma control was determined by the asthma control test (ACT) questionnaire. Asthma exacerbation was evaluated according to the criteria of the American Thoracic Society/European Respiratory Society. Levels of AMP, ACh, and HA in sputum were measured by liquid chromatography coupled to tandem mass spectrometry. IL-β, IL-4, IL-5, IL-6, IL-8, IL-13, IL-17A, TNF-α, IFN-γ, and macrophage-derived chemokine (MDC) were also measured.. Compared to healthy controls, asthmatics had higher levels of HA, lower levels of ACh, and similar levels of AMP in induced sputum samples. Compared to controlled asthma (n = 54), uncontrolled asthma (n = 58) showed higher AMP levels (P = 0.002), but similar HA and ACh levels. AMP was negatively correlated with ACT scores (r = - 0.348) and asthma quality of life questionnaire scores (r = - 0.188) and positively correlated with blood monocytes percentage (r = 0.195), sputum MDC (r = 0.214), and IL-6 levels (r = 0.196). Furthermore, AMP was associated with an increased risk of exacerbations in the preceding year.. Endogenous AMP, but not ACh or HA, was associated with asthma control, quality of life, and exacerbations in the previous year, which indicates that AMP could be a clinically useful biomarker of asthma.

Topics: Acetylcholine; Adenosine; Adenosine Monophosphate; Asthma; Biomarkers; Chemokine CCL22; Histamine; Humans; Interleukin-13; Interleukin-17; Interleukin-4; Interleukin-5; Interleukin-6; Interleukin-8; Quality Control; Quality of Life; Sputum; Tumor Necrosis Factor-alpha

Doublecortin-like kinase 1 (DCLK1) has been recognized as a marker of cancer stem cell in several malignancies. Thrombin is crucial in asthma severity as it can promote IL-8/CXCL8 production in lung epithelial cells, which is a potent chemoattractant for neutrophils. However, the pathologic role of DCLK1 in asthma and its involvement in thrombin-stimulated IL-8/CXCL8 expression remain unknown.. IL-8/CXCL8, thrombin, and DCLK1 expression were observed in the lung tissues of severe asthma patients and ovalbumin (OVA)-induced asthmatic mice model. A549 and BEAS-2B cells were either pretreated with inhibitors or small interfering RNAs (siRNAs) before being treated with thrombin. IL-8/CXCL8 expression and the molecules involved in signaling pathway were performed using ELISA, luciferase activity assay, Western blot, or ChIP assay.. IL-8/CXCL8, thrombin, and DCLK1 were overexpressed in the lung tissues of severe asthma patients and ovalbumin (OVA)-induced asthmatic mice model. Our in vitro study found that DCLK siRNA or LRKK2-IN-1 (DCLK1 inhibitor) attenuated IL-8/CXCL8 release after thrombin induction in A549 and BEAS-2B cells. Thrombin activated DCLK1, RhoA, and YAP in a time-dependent manner, in which DCLK1 siRNA inhibited RhoA and YAP activation. YAP was dephosphorylated on the Ser127 site after thrombin stimulation, resulting in YAP translocation to the nucleus from the cytosol. DCLK1, RhoA and YAP activation following thrombin stimulation were inhibited by U0126 (ERK inhibitor). Moreover, DCLK1 and YAP siRNA inhibited κB-luciferase activity. Thrombin stimulated the recruitment of YAP and p65 to the NF-κB site of the IL-8/CXCL8 promoter and was inhibited by DCLK1 siRNA.. Thrombin activates the DCLK1/RhoA signaling pathway, which promotes YAP activation and translocation to the nucleus from the cytosol, resulting in YAP/p65 formation, and binding to the NF-κB site, which enhances IL-8/CXCL8 expression. DCLK1 might be essential in thrombin-stimulated IL-8/CXCL8 expression in asthmatic lungs and indicates a potential therapeutic strategy for severe asthma treatment.

Topics: Animals; Asthma; Doublecortin-Like Kinases; Epithelial Cells; Humans; Interleukin-8; Luciferases; Lung; Mice; NF-kappa B; Ovalbumin; Phosphorylation; Protein Serine-Threonine Kinases; rhoA GTP-Binding Protein; RNA, Small Interfering; Thrombin

Maternal infection and stress during the prenatal period have been associated with adverse neurodevelopmental outcomes in offspring, suggesting that biomarkers of increased inflammation in the mothers may associate with poorer developmental outcomes. In 491 mother-child pairs from the Vitamin D Antenatal Asthma Reduction Trial (VDAART), we investigated the association between maternal levels of two inflammatory biomarkers; interleukin-8 (IL-8) and C-Reactive Protein (CRP) during early (10-18 wks) and late (32-38 wks) pregnancy with offspring scores in the five domains of the Ages and Stages Questionnaire, a validated screening tool for assessing early life development. We identified a robust association between early pregnancy IL-8 levels and decreased fine-motor (β: -0.919, 95%CI: -1.425, -0.414,

Topics: Asthma; Biomarkers; C-Reactive Protein; Clinical Trials as Topic; Female; Humans; Interleukin-8; Male; Pregnancy; Prenatal Exposure Delayed Effects; Vitamin D; Vitamins

Asthma is characterized by chronic inflammation of the airway mucosa. As Eucommia ulmoides Oliv. leaf extract (ELE) has been known to have anti-inflammatory properties, herein, we investigated the effect of ELE on interleukin (IL-) 8 production in A549 cells, a human airway epithelial cell line. The addition of ELE 1 h before tumor necrosis factor-alpha (TNFα) stimulation inhibited IL-8 production by A549 cells in a concentration-dependent manner. The addition of geniposidic acid, the main component of ELE, also inhibited IL-8 production. To further investigate the mechanism by which ELE inhibits IL-8 production, the effect of ELE or geniposidic acid on TNFα-stimulated p38 phosphorylation was examined by Western blotting. After 30 min of TNFα stimulation, p38 phosphorylation was inhibited by the addition of ELE or geniposidic acid, suggesting that ELE inhibited IL-8 production in TNFα-stimulated A549 cells by suppressing one of the signal transducers of p38 phosphorylation. These results indicate that ELE can be used as an effective measure against asthma, particularly neutrophilic asthma.

Topics: A549 Cells; Anti-Inflammatory Agents; Asthma; Eucommiaceae; Humans; Inflammation; Interleukin-8; p38 Mitogen-Activated Protein Kinases; Phytotherapy; Plant Extracts; Plant Leaves; Tumor Necrosis Factor-alpha

It has been approved that neutrophils are responsible for many inflammatory lung diseases, such as acute respiratory distress syndrome, chronic obstructive pulmonary disease, and asthma. It is well documented that the CXC chemokine interleukin-8 (IL-8) plays a key role as a potent neutrophil recruiting and activating factor. Asthma is one of the most common major non-contagious diseases and has a substantial impact on the patient's quality of life. The current evidence suggests that asthma is a complex multifactorial disorder, and its etiology is increasingly attributed to interactions between genetic susceptibility, host factors, and environmental exposures. IL-8 plays an important role in respiratory diseases and is a known regulator of pulmonary inflammation and immunity, induced phagocytosis, and promoted angiogenesis. This study aimed to investigate the IL-8 gene expression in blood samples of bronchial asthma patients. Therefore, the blood samples were taken from two groups of participants, including the group of patients with asthma (n=100) in the age range of20-61years and the group of healthy individuals (n=50).The obtained results indicated that the expression of IL-8 mRNA in the group of asthma patients was three times higher than that in the group of healthy individuals. Therefore, it is suggested that the antagonism of IL-8 could be a potent therapeutic strategy in the treatment of asthma.

Topics: Asthma; Humans; Interleukin-8; Iraq; Neutrophils; Quality of Life

Topics: Asthma; Cell Differentiation; Cell Line; Cells, Cultured; Cytokines; Gene Expression Profiling; Humans; Hypersensitivity; Intercellular Adhesion Molecule-1; Interleukin-8; Lipopolysaccharides; Mucin 5AC; Proto-Oncogene Proteins c-ets; Respiratory Hypersensitivity; Respiratory Mucosa; RNA, Antisense; RNA, Long Noncoding; Up-Regulation

Detection of asthma by a suitable biomarker is mandatory for the early identification, which helps in providing a right medication for the complete cure. Interleukins (ILs) have played a major role in asthma; in particular IL-8 is highly correlated with severe asthma. This research was focused on to detect IL-8 level by its partner antibody on a microgapped dual electrodes sensor. The sensing surface was modified into graphene oxide (GO), and an antibody was fixed by using the amine-aldehyde linker. GO enhanced the antibody immobilization and the consequence electric current flow upon interacting with IL-8. The detection limit of IL-8 was reached to 10 pg/mL in a linear range from 1 to 10,000 pg/mL with the regression of y = 0.7246x - 0.906 (R² = 0.9758); further, the sensitivity falls at 1 pg/mL. The surface does not show the antifouling effect with control antibody, and proteins, indicating the specific IL-8 detection. The detection of IL-8 helps in diagnosing and solving the related problems of asthmatic patients.

Topics: Asthma; Biosensing Techniques; Electrochemical Techniques; Electrodes; Graphite; Humans; Interleukin-8; Surface Properties

Asthma is a heterogeneous disease characterized by multiples respiratory symptoms; this is a polygenic entity that involves a complex interaction of environmental factors and inherent to the individual. To understand the development of asthma, some phenotypes have been proposed.. This work's purpose was to explore different molecules related to asthma development and to define each phenotype's specific characteristics.. 96 adult patients diagnosed with asthma before any treatment were enrolled in the protocol. Spirometric parameters, circulating leukocytes, serum IgE, body mass index, exhaled nitric oxide (FENO), and leukotrienes (LTB4) in urine were determined in each patient. The presence of asthma phenotypes proposed by the Global Initiative for Asthma (GINA) were explored: A) Allergic asthma, B) Non-allergic asthma, C) Late-onset asthma, D) Asthma with persistent airflow limitation, and E) Asthma with overweight and obesity.. In the cohort analyzed, we found four of phenotypes proposed by GINA; however, these phenotypes overlapped, due to this, 4 groups were integrated with allergic, non-allergic and obese patients, which were the main phenotypes. The main overlap was that of patients not-obese allergic, and was characterized by earlier onset, elevated levels of IgE, LTB4 and inflammasome related cytokines. Non-allergic patients had a significant association between interleukin (IL)-18 and IL-18 binding protein (BP) with narrow ratio between these cytokines. Finally, LTB4 had remarkable capacity to discriminate between allergic and not allergic patients.. Asthmatic phenotypes exist as interrelated characteristics and not as discrete entities. High levels of leukotrienes and IgE are hallmarks in the allergic phenotype of asthma.

Topics: Adult; Age of Onset; Asthma; Biomarkers; Cytokines; Eosinophils; Female; Humans; Hypersensitivity; Immunoglobulin E; Inflammasomes; Inflammation Mediators; Intercellular Signaling Peptides and Proteins; Interleukin-18; Interleukin-8; Leukotrienes; Male; Middle Aged; Overweight; Phenotype; Transforming Growth Factor beta

Oroscomucoid 3 (ORMDL3) has been linked to susceptibility of childhood asthma and respiratory viral infection. Polyinosinic-polycytidylic acid (poly I:C) is a synthetic analog of viral double-stranded RNA, a toll-like receptor 3 (TLR3) ligand and mimic of viral infection.. To investigate the functional role of ORMDL3 in the poly I:C-induced inflammatory response in airway epithelial cells, ORMDL3 knockdown and over-expression models were established in human A549 epithelial cells and primary normal human bronchial epithelial (NHBE) cells. The cells were stimulated with poly I:C or the Th17 cytokine IL-17A. IL-6 and IL-8 levels in supernatants,  mRNA levels of genes in the TLR3 pathway and inflammatory response from cell pellets were measured. ORMDL3 knockdown models in A549 and BEAS-2B epithelial cells were then infected with live human rhinovirus (HRV16) followed by IL-6 and IL-8 measurement.. ORMDL3 knockdown and over-expression had little influence on the transcript levels of TLR3 in airway epithelial cells. Time course studies showed that ORMDL3-deficient A549 and NHBE cells had an attenuated IL-6 and IL-8 response to poly I:C stimulation. A549 and NHBE cells over-expressing ORMDL3 released relatively more IL-6 and IL-8 following poly I:C stimulation. IL-17A exhibited a similar inflammatory response in ORMDL3 knockdown and over-expressing cells, but co-stimulation of poly I:C and IL-17A did not significantly enhance the IL-6 and IL-8 response. Transcript abundance of IFNB following poly I:C stimulation was not significantly altered by ORMDL3 knockdown or over-expression. Dampening of the IL-6 response by ORMDL3 knockdown was confirmed in HRV16 infected BEAS-2B and A549 cells.. ORMDL3 regulates the viral inflammatory response in airway epithelial cells via mechanisms independent of the TLR3 pathway.

Topics: A549 Cells; Asthma; Bronchi; Epithelial Cells; Humans; Interleukin-17; Interleukin-6; Interleukin-8; Membrane Proteins; Poly I-C; Respiratory Mucosa; RNA Interference; Toll-Like Receptor 3; Virus Diseases

Asthma is a common chronic illness among school children, where different cytokines, including IL-8 play a role in its pathogenesis. IL-8 induces chemotaxis and migration of immune cells, especially neutrophils to the site of inflammation. IL-8 level was significantly increased in sputum of severely asthmatic patients, but can it be linked to some asthma phenotypes. Our aim of the study was to detect the IL 8 gene expression in different asthma phenotypes and to determine its relation to asthma severity. This case control study included 320 subjects (160 asthmatic and 160 matched controls) aged from 5 to 16 years old in Beni-Suef governorate. IL-8 gene expression was assessed by a real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and studied regarding its level in cases versus controls and its relations to severity, phenotype and other laboratory parameters. IL-8 gene expression was statistically higher in asthmatic cases (P<0.001) and was significantly correlated to the phenotype (presence of other allergy as urticaria and drug eruption) and degree of asthma symptoms (r=0.869, P<0.001), FEV1(r=0.757, P<0.001) and serum IgE level (r=0.789, P<0.001). IL-8 gene expression level is increased with the degree of severity in asthmatic children and can be looked for in certain asthma phenotypes especially in presence of other atopic manifestation.

Topics: Adolescent; Aged; Asthma; Case-Control Studies; Child; Child, Preschool; Gene Expression; Humans; Interleukin-8; Phenotype

Early interactions between respiratory viruses and microbiota might modulate host immune responses and subsequently contribute to later development of recurrent wheezing and asthma in childhood. We aimed to study the possible association between respiratory microbiome, host immune response, and the development of recurrent wheezing in infants with severe respiratory syncytial virus (RSV) bronchiolitis.. Seventy-four infants who were hospitalized at Beijing Children's Hospital during an initial episode of severe RSV bronchiolitis at 6 months of age or less were included and followed up until the age of 3 years. Sputum samples were collected, and their microbiota profiles, LPS, and cytokines were analyzed by 16S rRNA-based sequencing, ELISA, and multiplex immunoassay, respectively.. Twenty-six (35.1%) infants developed recurrent wheezing by the age of 3 years, and 48 (64.9%) did not. The relative abundance of Haemophilus, Moraxella, and Klebsiella was higher in infants who later developed recurrent wheezing than in those who did not (LDA score >3.5). Airway levels of LPS (P = .003), CXCL8 (P = .004), CCL5 (P = .029), IL-6 (P = .004), and IL-13 (P < .001) were significantly higher in infants who later developed recurrent wheezing than in those who did not. Moreover, high airway abundance of Haemophilus was associated with CXCL8 (r = 0.246, P = .037) level, and that of Moraxella was associated with IL-6 level (r = 0.236, P = .046) and IL-10 level (r = 0.266, P = .024).. Our study suggests that higher abundance of Haemophilus and Moraxella in airway microbiome might modulate airway inflammation during severe RSV bronchiolitis in infancy, potentially contributing to the development of subsequent recurrent wheezing in later childhood.

Topics: Asthma; Beijing; Bronchiolitis; Child, Preschool; Female; Humans; Immunity; Infant; Interleukin-10; Interleukin-13; Interleukin-8; Male; Microbiota; Prospective Studies; Recurrence; Respiratory Sounds; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses; Respiratory System; RNA, Ribosomal, 16S; Sputum

Thrombin plays a crucial role in lung inflammatory diseases such as asthma and chronic obstructive pulmonary disease (COPD). Thrombin induces the release of interleukin-8 (IL-8)/CXCL8 by lung epithelial cells, and this phenomenon plays a vital role in lung inflammation. Our previous studies have indicated that thrombin stimulates IL-8/CXCL8 expression through PI3K/Akt/IκB kinase (IKK)α/β/nuclear factor-κB (NF-κB) and p300 pathways in human lung epithelial cells. In the present study, we explored the roles of mammalian target of rapamycin (mTOR) and p70S6 kinase (p70S6K) in thrombin-induced NF-κB activation and IL-8/CXCL8 release in human lung epithelial cells. In this study, we found that rapamycin (an mTOR inhibitor) and p70S6K siRNA diminished thrombin-induced IL-8/CXCL8 release. Thrombin induced mTOR Ser2448 phosphorylation and p70S6K Thr389 phosphorylation in a time-dependent manner. Moreover, rapamycin attenuated thrombin-stimulated p70S6K phosphorylation. We also found that transfection of cells with the dominant negative mutant of Akt (Akt DN) reduced the thrombin-induced increase in mTOR phosphorylation and p70S6K phosphorylation. Moreover, thrombin-stimulated p300 phosphorylation was attenuated by Akt DN, rapamycin, and p70S6K siRNA. Thrombin triggered p70S6K translocation from the cytosol to the nucleus in a time-dependent manner. Thrombin induced the complex formation of p70S6K, p300, and p65; acetylation of p65 Lys310, and recruitment of p70S6K, p300, and p65 to the κB-binding site of the IL-8/CXCL8 promoter region. In conclusion, these results indicate that thrombin initiates the Akt-dependent mTOR/p70S6K signaling pathway to promote p300 phosphorylation and NF-κB activation and finally induces IL-8/CXCL8 release in human lung epithelial cells.

Topics: A549 Cells; Asthma; E1A-Associated p300 Protein; Epithelial Cells; Humans; Interleukin-8; Lung; Phosphorylation; Pulmonary Disease, Chronic Obstructive; Ribosomal Protein S6 Kinases, 70-kDa; Signal Transduction; Thrombin; TOR Serine-Threonine Kinases; Transcription Factor RelA

Some patients with asthma present with accelerated lung function decline. This phenomenon is mostly associated with severe exacerbations and with poor asthma control.. Our aim was to detect the extent of FEV1 decline in patients with mild asthma and to discriminate clinical, functional and inflammatory factors associated with accelerated FEV1 decline.. We recruited 50 patients with mild asthma for pulmonary function testing and induced sputum sampling 12-15 years after the initial diagnosis. In 33 patients, from whom sputum of a good quality was obtained, inflammatory cells were counted and concentrations of cytokines IL-2, IL-4, IL-5, IL-8, IL-10, IFN-γ, angiogenin and VEGF in the sputum were measured by cytometric bead array.. Eighteen of 33 patients presented with accelerated FEV1 decline of more than 30 ml/year, with a mean (SEM) of 43.2 (3.9) ml/year, compared to 15 control patients with a FEV1 decline of 14.4 (2.1) ml/year. In the accelerated FEV1 decline group, we found elevated sputum levels of IL5 with a median (IQR) of 1.8 (0.4-3.2) pg/ml vs. 0.2 (0.1-1.2) pg/ml, p = 0.04; IL8 with a mean (SEM) of 1503 (194) pg/ml vs. 938 (177) pg/ml, p = 0.04; and eosinophils with a median (IQR) of 223 (41-1020) cells/μl vs. 39 (1-190) cells/μl, p = 0.03. No significant differences in other measured parameters were detected between the two groups.. Elevated sputum eosinophils, IL5 and IL8, which have a potential to stimulate airway remodelling, might be a useful non-invasive biomarkers and therapeutic targets of accelerated FEV1 decline in asthma patients.

Topics: Asthma; Disease Progression; Eosinophils; Female; Forced Expiratory Volume; Humans; Inflammation Mediators; Interleukin-5; Interleukin-8; Male; Middle Aged; Severity of Illness Index; Sputum

To distinguish between first wheeze and asthma in early childhood, we investigated respiratory viruses and cytokine/chemokine profiles among patients with first wheeze and established asthma.. We enrolled children with acute exacerbations of wheezing (17 first wheeze and 32 asthma) and 11 controls (no wheezing) aged between 10 months and 6 years. Nasal aspirates were obtained, and virus detection was performed with antigenic assay kits and/or RT-PCR. Serum 27 cytokines/chemokines were assayed by a multi-cytokine detection system.. Rhinovirus and respiratory syncytial (RS) virus were dominant in acute exacerbations of asthma. However, many types of viruses were isolated in first wheeze. Serum IL-8 and IL-12 values were significantly higher in first wheeze than in acute asthma or the controls. IL-5 and IP-10 levels in acute asthma and first wheeze cases were higher than in the controls. Both of them were significantly higher in cases of acute asthma than in convalescence stage of asthma cases. Only IP-10 was significantly higher in first wheeze than in convalescence stage of first wheeze cases.. Different profiles in virus detection and production of IL-8 and IL-12 might distinguish between first wheeze and childhood asthma.

Topics: Asthma; Child; Child, Preschool; Disease Progression; Eosinophils; Female; Humans; Infant; Interleukin-12; Interleukin-8; Leukocyte Count; Male; Neutrophils; Respiratory Sounds; Respiratory Syncytial Viruses; Rhinovirus

The 2015 definition for pediatric acute respiratory distress syndrome did not require the presence of bilateral infiltrates. We tested the hypothesis that pediatric patients meeting oxygenation criteria for pediatric acute respiratory distress syndrome but without bilateral infiltrates would have different inflammatory biomarker levels and clinical outcomes than those with bilateral infiltrates.. Secondary analysis of a prospective cohort study.. Twenty-two PICUs.. Four-hundred forty-six patients age 2 weeks to 17 years intubated for respiratory failure with oxygenation index greater than or equal to 4 or oxygenation saturation index greater than or equal to 5 on the day of intubation or the day after.. None.. Patients with bilateral infiltrates, either on the day of intubation or within the following 2 days, were compared with children who never developed bilateral infiltrates. Two analyses were performed to test 1) whether bilateral infiltrates are associated with elevated interleukin-1 receptor antagonist or interleukin-8 and 2) whether bilateral infiltrates are associated with worse clinical outcomes. Patients with bilateral infiltrates more often had a primary diagnosis of pneumonia (41% vs 28%; p = 0.02) and less often asthma (8% vs 23%; p < 0.01). After controlling for age, gender, and primary diagnosis, interleukin-1 receptor antagonist was higher on study days 1 and 2 in patients with bilateral infiltrates. There was no difference in interleukin-8 levels. After adjusting for age, gender, Pediatric Risk of Mortality score, and severity of oxygenation defect, presence of bilateral infiltrates was associated with longer duration of mechanical ventilation in survivors (hazard ratio, 0.64; 95% CI, 0.49-0.82; p < 0.01); this association was independent of primary diagnosis. Overall mortality was 9%; mortality was higher in those without bilateral infiltrates (14% vs 8%; p = 0.04).. Children meeting pediatric acute respiratory distress syndrome oxygenation criteria with bilateral infiltrates on chest radiograph experience a more intense early inflammatory response. Bilateral infiltrates are associated with longer time on the ventilator independent of oxygenation defect severity.

Topics: Adolescent; Asthma; Biomarkers; Child; Child, Preschool; Female; Humans; Infant; Infant, Newborn; Inflammation Mediators; Intensive Care Units, Pediatric; Interleukin-8; Intubation, Intratracheal; Male; Prospective Studies; Receptors, Interleukin-1; Respiration, Artificial; Respiratory Distress Syndrome; Risk Factors; Severity of Illness Index

The incidence of obesity-related asthma has shown a remarkable increase.. We aimed to explore the role of heat shock protein 72 (Hsp72) and receptor for advanced glycation end products (RAGE) axis with its downstream signaling in the pathogenesis of obesity-related asthma.. We enrolled a total of 55 subjects and divided them into three groups. Groups I and II included healthy, normal weight (n = 15) and obese (n = 15) subjects, respectively. Twenty-five obese asthmatics (group III) were subdivided into group IIIa (10 patients with mild to moderate asthma) and group IIIb (15 patients with severe asthma). High mobility group box 1 (HMGB1), interleukin 8 (IL-8), monocyte chemoattractant protein 1 (MCP-1), extracellular signal-regulated protein kinases 1 and 2 (ERK1/2), and urinary Hsp72 were immunoassayed. Hydrogen peroxide (H. The current study highlights the role of Hsp72 and HMGB1/RAGE/ERK1/2 signaling cascade in the pathogenesis of bronchial asthma and its link to obesity, which could be reflected on monitoring, severity grading, and management of this disease.

Topics: Adult; Antigens, Neoplasm; Asthma; Case-Control Studies; Chemokine CCL2; Fatty Acids, Nonesterified; Female; Heat-Shock Proteins; HMGB1 Protein; Humans; Hydrogen Peroxide; Interleukin-8; Male; MAP Kinase Signaling System; Middle Aged; Mitogen-Activated Protein Kinases; Molecular Chaperones; Obesity; Receptor Cross-Talk

Topics: Administration, Inhalation; Adrenal Cortex Hormones; Angiotensin-Converting Enzyme 2; Anti-Asthmatic Agents; Asthma; Betacoronavirus; Coronavirus Infections; COVID-19; Cytokine Release Syndrome; Dexamethasone; Endoribonucleases; Gene Expression Regulation; Host-Pathogen Interactions; Humans; Interleukin-6; Interleukin-8; Pandemics; Peptidyl-Dipeptidase A; Pneumonia, Viral; Receptors, Virus; SARS-CoV-2; Serine Endopeptidases; Severity of Illness Index; Tumor Necrosis Factor-alpha; Viral Nonstructural Proteins

Rhinoviral infection is associated with an increased risk of asthma attacks. The macrolide clarithromycin decreases cytokine production in nasopharyngeal aspirates from patients with wheezing, but the effects of macrolides on cytokine production in nasal epithelial cells obtained from asthmatic subjects remain unclear. Here, human nasal epithelial cells were infected with type-14 rhinovirus (RV14), a major RV group. Titers and RNA of RV14 and cytokine concentrations, including IL-1β and IL-6, were higher in the supernatants of the cells obtained from subjects with bronchial asthma (asthmatic group) than in those from the non-asthmatic group. Pretreatment with clarithromycin decreased RV14 titers, viral RNA and cytokine concentrations, and susceptibility to RV14 infection. Pretreatment with clarithromycin also decreased IL-33 production, which was detected after infection. Pretreatment with clarithromycin decreased the expression of intercellular adhesion molecule-1, the receptor for RV14, after infection, the number and fluorescence intensity of the acidic endosomes through which RV RNA enters the cytoplasm, and the activation of nuclear factor kappa-B proteins in nuclear extracts. These findings suggested that RV replication and cytokine production may be enhanced in nasal epithelial cells obtained from subjects with bronchial asthma and may be modulated by clarithromycin.

Topics: Antiviral Agents; Asthma; Cells, Cultured; Clarithromycin; Cytokines; Epithelial Cells; Female; Humans; Interleukin-33; Interleukin-6; Interleukin-8; Male; Middle Aged; Rhinovirus; Virus Replication

Topics: Asthma; beta Catenin; Cells, Cultured; Chemokine CCL8; Cytokines; Frizzled Receptors; Humans; Interleukin-8; Mast Cells; Receptors, Wnt; Signal Transduction; Transcriptome; Wnt Signaling Pathway; Wnt3A Protein

Variability in response to inhaled corticosteroids (ICSs) can result in less than optimal asthma control. Development of biomarkers assessing the therapeutic efficacy of corticosteroids is important.. We sought to examine whether in vitro PBMC responses to corticosteroids relate to the clinical ICS response.. Compared with PBMCs from patients with easy-to-control asthma, PBMCs from those with difficult-to-control asthma had significantly lower glucocorticoid receptor α levels at V0 (P = .05). A 30% increase in IL-8 suppression by FLU (P = .04) and a trend for increased TNF-α suppression by FLU between V0 and V6 (P = .07) were observed in patients with easy-to-control asthma. In contrast, no changes between V0 and V6 in IL-8 and TNF-α suppression by FLU were observed in patients with difficult-to-control asthma. Corticosteroid-mediated transactivation (FK506-binding protein 5 induction by FLU) increased in the PBMCs of patients with difficult-to-control and easy-to-control asthma between V0 and V6 (P = .05 and P = .03, respectively).. PBMCs of children with difficult-to-control asthma treated with guidelines-based therapy and requiring high-dose ICSs had reduced in vitro responsiveness to corticosteroids.

Topics: Adolescent; Adrenal Cortex Hormones; Anti-Asthmatic Agents; Asthma; Cells, Cultured; Child; Female; Fluticasone; Gene Expression Regulation; Humans; Interleukin-8; Leukocytes, Mononuclear; Male; Receptors, Glucocorticoid; Tacrolimus Binding Proteins; Tumor Necrosis Factor-alpha; Vitamin D3 24-Hydroxylase

Polymorphisms on chromosome 17q21 confer the major genetic susceptibility to childhood-onset asthma. Risk alleles positively correlate with ORMDL3 (orosomucoid-like 3) expression. The locus influences disease severity and the frequency of human rhinovirus (HRV)-initiated exacerbations. ORMDL3 is known to regulate sphingolipid synthesis by binding serine palmitoyltransferase, but its role in inflammation is incompletely understood.. To investigate the role of ORMDL3 in cellular inflammation.. We modeled a time series of IL1B-induced inflammation in A549 cells, using cytokine production as outputs and testing effects of ORMDL3 siRNA knockdown, ORMDL3 overexpression, and the serine palmitoyltransferase inhibitor myriocin. We replicated selected findings in normal human bronchial epithelial cells. Cytokine and metabolite levels were analyzed by analysis of variance. Transcript abundances were analyzed by group means parameterization, controlling the false discovery rate below 0.05.. Silencing ORMDL3 led to steroid-independent reduction of IL6 and IL8 release and reduced endoplasmic reticulum stress after IL1B stimulation. Overexpression and myriocin conversely augmented cytokine release. Knockdown reduced expression of genes regulating host-pathogen interactions, stress responses, and ubiquitination: in particular, ORMDL3 knockdown strongly reduced expression of the HRV receptor ICAM1. Silencing led to changes in levels of transcripts and metabolites integral to glycolysis. Increased levels of ceramides and the immune mediator sphingosine-1-phosphate were also observed.. The results show ORMDL3 has pleiotropic effects during cellular inflammation, consistent with its substantial genetic influence on childhood asthma. Actions on ICAM1 provide a mechanism for the locus to confer susceptibility to HRV-induced asthma.

Topics: A549 Cells; Asthma; Cytokines; Endoplasmic Reticulum Stress; Gene Expression Profiling; Gene Knockdown Techniques; Humans; Inflammation; Interleukin-6; Interleukin-8; Membrane Proteins; Sphingolipids

Steroid-insensitive asthma-related airway inflammation is associated with the expression of epidermal growth factor receptor (EGFR) tyrosine kinase in asthmatic bronchial epithelium. Proinflammatory cytokines IL-6 and IL-8 are related to steroid-insensitive asthma. It is currently unknown how EGFR-tyrosine kinase inhibitors (EGFR-TKIs) affects house dust mite (HDM)-induced asthma in terms of inflammatory cytokines related to steroid-resistant asthma and further signaling pathway. Cytokine expressions and EGFR signaling pathway were performed by ELISA, reverse transcriptase PCR, real-time PCR, and Western blot in cell-line models. AMP-activated protein kinase (AMPK) pathway-related inhibitors were applied to confirm the association between EGFR-TKI and AMPK pathway. HDM induced IL-6 and IL-8 in a dose-dependent manner. Both Erlotinib (Tarceva) and Osimertinib (AZD-9291) reduced the levels of HDM-stimulated IL-6 and IL-8 levels in BEAS-2B cells. AZD-9291 was more effective than Erlotinib in inhibiting phospho-EGFR, and downstream phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) and phopho-signal transducer and activator of transcription 3 (p-STAT3) pathway signaling. In addition, AMPK pathway-related inhibitor, Calcium-/calmodulin-dependent protein kinase kinase β (CaMKKβ) inhibitor, down-regulated IL-8, but EGFR-TKI had no effect on AMPK pathway. Our findings highlight EGFR-TKIs, Tarceva, and AZD-9291, attenuate HDM-induced inflammatory IL-6 and IL-8 cytokines via EGFR signaling axis pathway, but not AMPK signaling pathway.

Topics: Acrylamides; Aniline Compounds; Animals; Antigens, Dermatophagoides; Arthropod Proteins; Asthma; Cell Line; Dermatophagoides pteronyssinus; Dose-Response Relationship, Drug; Epithelial Cells; ErbB Receptors; Erlotinib Hydrochloride; Humans; Interleukin-6; Interleukin-8; Respiratory Mucosa; Signal Transduction

Topics: Acute Disease; Adult; Asthma; Blotting, Western; Case-Control Studies; DNA; Extracellular Traps; Female; Glucosephosphate Dehydrogenase; Humans; Inflammasomes; Interleukin-6; Interleukin-8; Longitudinal Studies; Male; Middle Aged; Neutrophils

Topics: Airway Obstruction; Asthma; Biomarkers; Bronchial Diseases; Comorbidity; Cross-Sectional Studies; Eosinophilia; Female; Glucocorticoids; Goals; Humans; Interleukin-8; Male; Medication Adherence; Middle Aged; Nebulizers and Vaporizers; Severity of Illness Index; Spain

Neutrophilic inflammation in asthma is associated with interleukin (IL)-17A, corticosteroid-insensitivity and bronchodilator-induced forced expiratory volume in 1 s (FEV

Topics: Adolescent; Adrenal Cortex Hormones; Adult; Aged; Albuterol; Asthma; Bronchi; Bronchial Hyperreactivity; Bronchoalveolar Lavage; Cell Line, Tumor; Cytoplasm; Epithelial Cells; Epithelium; Female; Forced Expiratory Volume; Humans; Inflammation; Interleukin-17; Interleukin-8; Lung; Male; Methacholine Chloride; Middle Aged; Neutrophils; Randomized Controlled Trials as Topic; Respiratory Function Tests; Smoking; Tumor Necrosis Factor-alpha; Young Adult

Asthma is a complex disease comprising various phenotypes and endotypes, all of which still need solid biomarkers for accurate classification. In a previous study, we defined specific genes related to asthma and respiratory allergy by studying the expression of 94 genes in a population composed of 4 groups of subjects: healthy control, nonallergic asthmatic, asthmatic allergic, and nonasthmatic allergic patients. An analysis of differential gene expression between controls and patients revealed a set of statistically relevant genes mainly associated with disease severity, i.e.,

Topics: Adult; Aged; Asthma; Biomarkers; Chitinase-3-Like Protein 1; Diagnosis, Differential; Disease Progression; Female; Humans; Hypersensitivity; Interleukin-10; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Phenotype; Sensitivity and Specificity

Bronchial asthma (BA) is a common chronic respiratory disease that has exhibited a rising global incidence in recent years. Glucocorticoids are used for the treatment of BA. Emerging evidence has demonstrated the roles of tumor necrosis factor (TNF-α), interleukin-8 (IL-8) and eosinophil cationic protein (ECP) in BA. The present study investigated whether TNF-α, IL-8 and ECP were associated with the clinical stages and severity of BA and the efficacy of glucocorticoids in the treatment of BA.. A total of 199 patients with BA and 174 healthy individuals were included in this study. Patients with BA underwent glucocorticoid treatment, and the TNF-α, IL-8 and ECP levels and lung functions of the subjects were measured. The correlations of the TNF-α, IL-8 and ECP levels with BA severity, clinical staging and lung functions were assessed. We investigated whether the TNF-α, IL-8 and ECP levels aided in evaluating the efficacy of using glucocorticoids for the treatment of BA.. TNF-α, IL-8 and ECP exhibited high levels in patients with BA, and glucocorticoid treatment notably decreased these levels. The TNF-α, IL-8 and ECP levels were positively correlated with the clinical stages and severity of BA and negatively correlated with lung function. TNF-α, IL-8 and ECP can be used as serum markers to predict the efficacy of glucocorticoids in the treatment of BA.. The key findings of this study collectively support a role for TNF-α, IL-8 and ECP in BA development, and TNF-α, IL-8 and ECP can be used as serum markers of glucocorticoid efficacy in BA.

Topics: Adult; Analysis of Variance; Asthma; Eosinophil Cationic Protein; Female; Forced Expiratory Volume; Glucocorticoids; Humans; Interleukin-8; Male; Middle Aged; ROC Curve; Severity of Illness Index; Tumor Necrosis Factor-alpha; Vital Capacity

Chemokine CCL14 is inactive in its proform. Here, we show that inflammation- and cancer-associated kallikrein-related peptidases KLK5 and KLK8 remove the N-terminal eight amino acids from the proform thereby converting CCL14 to its active state. Activity of the chemokine is demonstrated by migration of myeloid cells expressing relevant receptors.

Topics: Asthma; Atherosclerosis; Cell Line, Tumor; Chemokine CX3CL1; Chemokine CXCL12; Chemokines; Chemokines, CC; Crohn Disease; Enzyme Activation; Humans; Interleukin-8; Kallikreins; Leukemia; Macrophage Inflammatory Proteins; Pancreatitis; Reactive Oxygen Species

In children with asthma, the viral infection of airways is usually a main cause of acute asthma exacerbation and hospitalization. However, few studies on clinical and biomolecular characteristics of asthmatic children in this field have been done, especially in emergent countries.. This study described the clinical and biological characteristics of asthmatic children who had acute asthma exacerbation and rhinovirus (RV) infection.. Children under 15 years of age hospitalized for acute asthma exacerbation were included. They underwent clinical examination and peripheral blood analyses for the cytokine profile. The severity of acute asthma exacerbation was evaluated by Pediatric Asthma Score (PAS). Healthy children under 15 years of age were also invited in this study.. One hundred fifteen asthmatic children were included in this study. There were 18.2% of mild PAS, 37.4% of moderate PAS, and 44.4% of severe PSA. Among them, 63/115 (54.8%) asthmatic children had positive RV infection (RV. RV infection is a main cause of acute asthma exacerbation in children with asthma. The increase of Th2-related cytokines, especially IL-5 and IL-13, is a relevant biomarker for RV infection in asthmatic children with severe exacerbation.

Topics: Adolescent; Asthma; Case-Control Studies; Child; Child, Preschool; Cytokines; Disease Progression; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Hospitals, Pediatric; Humans; Infant; Interferon-gamma; Interleukin-10; Interleukin-13; Interleukin-2; Interleukin-4; Interleukin-5; Interleukin-8; Male; Neutrophils; Picornaviridae Infections; Respiratory Tract Infections; Rhinovirus; Th1 Cells; Th2 Cells; Tumor Necrosis Factor-alpha

Orosomucoid like-3 (ORMDL3) has been identified to be associated with the development of asthma according to previous studies. However, the definite role of ORMDL3 in the pathogenesis of asthma remains unclear. In this study, we found ORMDL3 was highly expressed in PBMC specimens from childhood asthma patients. Cytokines production and p-ERK/MMP-9 pathway expression was also increased in childhood asthma patients compared with controls. In addition, ORMDL3 overexpression induced IL-6 and IL-8 release and activated p-ERK/MMP-9 pathway in vitro. Increased ORMDL3 expression was observed after treated with 5-Aza-CdR. 5-Aza-CdR decreased the percentage of the CpG island in the ORMDL3 promoter region and increased its promoter activity. In addition, 5-Aza-CdR significantly increased IL-6 and IL-8 levels in NHBE cells while there was no obvious alteration after knocking down ORMDL3. Knockdown of ORMDL3 also significantly decreased the expression of p-ERK/MMP-9 pathway in the presence or absence of 5-Aza-CdR. In conclusion, our study provided novel evidence for the association between ORMDL3 and asthma-associated cytokines. Moreover, DNA methylation plays an important role in ORMDL3-mediated increased IL-6 and IL-8 levels and p-ERK/MMP-9 pathway expression.

Topics: Adolescent; Asthma; Base Sequence; Case-Control Studies; Cell Line, Transformed; Child; CpG Islands; Decitabine; Epigenesis, Genetic; Epithelial Cells; Female; Humans; Interleukin-6; Interleukin-8; Male; Matrix Metalloproteinase 9; Membrane Proteins; Methylation; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Promoter Regions, Genetic; Respiratory Mucosa; Signal Transduction

Human airway smooth muscle cells (ASMCs) may have a pro-inflammatory role through the release of inflammatory mediators. Increasing evidence indicates that human β-defensins (HBDs) are related to pathogenesis of asthma.. To examine the plasma level of HBD-1, HBD-2 and HBD-3 in asthmatic patients and the expression of their mouse orthologues in the lung tissue of a mouse model of chronic severe asthma. Further to investigate the effect of HBD-3 on the release of the pro-inflammatory cytokine IL-8 and to explore the mechanisms.. The plasma levels of HBD-1, HBD-2 and HBD-3 from 34 healthy controls and 25 asthmatic patients were determined by ELISA. The expression of mouse β-defensins MBD-1, MBD-3 and MBD-14 in the lung tissue of asthmatic mice was detected by Western blot. The ASMCs were cultured with HBD-3 for 24 hour, and then the supernatant level of IL-8 was evaluated by ELISA and the cell viability was examined by WST-1 assay. The signalling pathway was investigated with blocking antibodies or pharmacological inhibitors.. The plasma levels of HBD-1 and HBD-3 were elevated in asthmatic patients, and the expression of MBD-14, the mouse orthologue for HBD-3, was increased in asthmatic mice. HBD-3-induced IL-8 production in a CCR6 receptor-specific manner and was dependent on multiple signalling pathways. Moreover, HBD-3-induced cell apoptosis concurrently, which was dependent on the ERK1/2 MAPK pathway. Mitochondrial ROS regulated both HBD-3-induced IL-8 production and cell apoptosis.. These observations provide clear evidence of an important new mechanism for the promotion of airway inflammation and tissue remodelling with potential relevance for the treatment of asthma.

Topics: Allergens; Animals; Apoptosis; Asthma; beta-Defensins; Biomarkers; Case-Control Studies; Disease Models, Animal; Humans; Interleukin-8; MAP Kinase Signaling System; Mice; Mitochondria, Muscle; Models, Biological; Myocytes, Smooth Muscle; NF-kappa B; Reactive Oxygen Species; Receptors, CCR6; Respiratory System; Signal Transduction

The extracellular matrix (ECM) creates the microenvironment of the tissue; an altered ECM in the asthmatic airway may be central in airway inflammation and remodelling. Tumstatin is a collagen IV-derived matrikine reduced in the asthmatic airway wall that reverses airway inflammation and remodelling in small and large animal models of asthma. This study hypothesized that the mechanisms underlying the broad asthma-resolving effects of tumstatin were due to autocrine remodelling of the ECM. Neutrophils and endothelial cells were seeded on decellularized ECM of non-asthmatic (NA) or asthmatic (A) airway smooth muscle (ASM) cells previously exposed to tumstatin in the presence or absence of a broad matrix metalloproteinase inhibitor, Marimastat. Gene expression in NA and A ASM induced by tumstatin was assessed using RT-PCR arrays. The presence of tumstatin during ECM deposition affected neutrophil and endothelial cell properties on both NA and A ASM-derived matrices and this was only partly due to MMP activity. Gene expression patterns in response to tumstatin in NA and A ASM cells were different. Tumstatin may foster an anti-inflammatory and anti-angiogenic microenvironment by modifying ASM-derived ECM. Further work is required to examine whether restoring tumstatin levels in the asthmatic airway represents a potential novel therapeutic approach.

Topics: Airway Remodeling; Angiogenesis Inhibitors; Anti-Inflammatory Agents, Non-Steroidal; Asthma; Autoantigens; Bronchi; Chemotaxis; Collagen Type IV; Enzyme Inhibitors; Extracellular Matrix; Gene Expression Profiling; Gene Expression Regulation; Human Umbilical Vein Endothelial Cells; Humans; Hydroxamic Acids; Interleukin-8; Matrix Metalloproteinases; Myocytes, Smooth Muscle; Neutrophils

Topics: Adult; Asthma; Biomarkers; Case-Control Studies; Estradiol; Female; Follicular Phase; Humans; Inflammation Mediators; Interleukin-5; Interleukin-8; Luteal Phase; Middle Aged; Progesterone; Prospective Studies; Sputum; Testosterone; Up-Regulation; Young Adult

To examine the IL-8 expression levels and association of genetic variants with the risk of childhood persistent asthma prognosis.. Overall, 170 asthmatic children and 170 healthy controls were included in this case-control study. The human IL-8 serum levels were measured using ELISA. The IL-8 mRNA expression levels were assessed by a real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods.. The IL-8 expression at both protein and mRNA levels was found to be significantly elevated in asthmatic children compared to healthy subjects (P < 0.0001, P = 0.004; respectively). Higher levels of IL-8 mRNA are detected in subjects with moderate to severe asthma. The presence of IL8-251 A/T (rs4073) and + 781C/T (rs2227306) polymorphisms was significantly associated with an increased risk of asthma (P = 0.002, P = 0.036, respectively). In addition, we noted a significant association between these polymorphisms and an elevated risk of atopic asthma (P < 0.05). For rs2227306 SNP, the highest median level of IgE was detected for the presence of TT genotype (865 ± 99.74 IU/mL). Although, the rs4073 polymorphism conferred a higher risk to develop asthma at an advanced stage of severity (P = 0.008). The rs4073 T and rs2227306 C alleles are considered as risk factors for asthma development. The rs4073 T allele is represented also as a risk factor for asthma severity in Tunisian children.. Both IL-8 gene and protein expression may play a key role in asthma pathogenesis.

Topics: Adolescent; Alleles; Asthma; Case-Control Studies; Child; Child, Preschool; Female; Gene Expression; Haplotypes; Humans; Immunoglobulin E; Interleukin-8; Male; Polymorphism, Single Nucleotide; Prognosis; RNA, Messenger; Severity of Illness Index; Tunisia

Endoplasmic reticulum (ER) stress has been considered to be an important regulator of airway inflammation in the pathogenesis of bronchial asthma, but the mechanism of ER stress involved in neutrophilic asthma remain not fully understood.. Tunicamycin is a mixture of homologous nucleoside antibiotics, which is used to induce ER stress. In the present study, Tunicamycin was administered to mouse bronchial epithelial cells and a neutrophilic asthma model (OVA. Tunicamycin not only induced ER stress in mouse bronchial epithelial cells, but also increased expression of inflammation indicators such as IL-6, IL-8, and TNF-α via PERK-ATF4-CHOP signaling. Additionally, the phosphorylation of PERK and the expression levels of ATF4 and CHOP proteins and inflammatory cytokines (IL-6, IL-8 and TNF-α) were elevated in the lung tissue of OVA. These data support the emerging notion that regulation of ER stress could be strongly associated with the development of neutrophilic asthma.

Topics: Activating Transcription Factor 4; Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Cells, Cultured; Cytokines; Disease Models, Animal; eIF-2 Kinase; Endoplasmic Reticulum Stress; Epithelial Cells; Inflammation Mediators; Interleukin-6; Interleukin-8; Mice; Signal Transduction; Transcription Factor CHOP; Tumor Necrosis Factor-alpha; Tunicamycin

Short chain fatty acids (SCFAs) are produced following the fermentation of soluble fibre by gut bacteria. In animal models, both dietary fibre and SCFAs have demonstrated anti-inflammatory effects via the activation of free fatty acid receptors, such as G protein-coupled receptor 41 and 43 (GPR41 and GPR43). This pilot study examined the acute effect of a single dose of soluble fibre on airway inflammation-including changes in gene expression of free fatty acid receptors-in asthma. Adults with stable asthma consumed a soluble fibre meal (

Topics: Adult; Asthma; Body Mass Index; Body Weight; Dietary Fiber; Female; Humans; Inflammation; Interleukin-8; Inulin; Macrophages; Male; Meals; Middle Aged; Neutrophils; Nitric Oxide; Pilot Projects; Probiotics; Receptors, Cell Surface; Receptors, G-Protein-Coupled; Sputum; Up-Regulation

The innate immune response is impaired in asthma, with increased epithelial release of C-X-C motif chemokine ligand (CXCL)8, interleukin (IL)-33 and thymic stromal lymphopoietin (TSLP). We hypothesised that dendritic cells might modulate the hyperresponsive epithelium in severe asthma.For this purpose, we investigated epithelial-dendritic crosstalk in normal and diseased conditions, and because ultrafine particulate matter may affect asthmatic airways, we investigated its impact on this crosstalk. Air-liquid interface cultures of human bronchial epithelial cells (HBEC) of control subjects (cHBEC) or severe asthma patients (saHBEC) were co-cultured with monocyte-derived dendritic cells (moDC).Increased release of CXCL8, TSLP and IL-33 from saHBEC contrasted with cHBEC producing CXCL10 and CCL2. Regarding moDC activation, saHBEC co-cultures induced only upregulation of CD86 expression, while cHBEC yielded full moDC maturation with HLA-DR, CD80, CD86 and CD40 upregulation. Particulate matter stimulation of HBEC had no effect on cHBEC but stimulated CXCL8 and IL-33 release in saHBEC. Particulate matter impaired epithelium signalling (TSLP, IL-33 and CXCL8) in saHBEC co-cultures despite C-C chemokine ligand 2 induction.Crosstalk between HBEC and moDC can be established

Topics: Adult; Aged; Aged, 80 and over; Asthma; Cells, Cultured; Coculture Techniques; Cytokines; Dendritic Cells; Epithelial Cells; Female; Humans; Interleukin-33; Interleukin-8; Male; Middle Aged; Th2 Cells; Thymic Stromal Lymphopoietin

Major basic protein (MBP) derived from activated eosinophil can exacerbate atopic asthma induced by lipopolysaccharide (LPS). The pharmacological function of MBP can be mimicked by poly-L-arginine (PLA), however, the potential signaling mechanisms of LPS-PLA-induced release of the inflammatory cytokines interleukin (IL)-6 and IL-8 remain unclear. In the present study, airway epithelia NCI-H292 cell lines were treated with LPS and/or PLA. We found that the expression levels of IL-6 and IL-8 induced by LPS-PLA were increased significantly compared with that in untreated cells. Meanwhile, the phosphorylation of p38 MAPK and ERK1/2 was also up-regulated dramatically by LPS-PLA, but this increase could be blocked by specific inhibitor. Importantly, blocking the phosphorylation of p38 MAPK and ERK1/2 reduced the expression levels of IL-6 and IL-8 as well. Collectively, LPS-PLA-induced release of IL-6 and IL-8 from NCI-H292 cells may be due to the synergistic activation of p38 MAPK and ERK1/2 signaling transduction pathways.

Topics: Asthma; Cell Line, Tumor; Enzyme Activation; Eosinophil Major Basic Protein; Extracellular Signal-Regulated MAP Kinases; Humans; Interleukin-6; Interleukin-8; Lipopolysaccharides; p38 Mitogen-Activated Protein Kinases; Peptides; Phosphorylation; Respiratory Mucosa

Respiratory viral infections are a major cause of asthma exacerbations. Neutrophils accumulate in the airways and the mechanisms that link neutrophilic inflammation, viral infections and exacerbations are unclear. This study aims to investigate anti-viral responses in neutrophils from patients with and without asthma and to investigate if neutrophils can be directly activated by respiratory viruses.. Neutrophils from peripheral blood from asthmatic and non-asthmatic individuals were isolated and stimulated with lipopolysaccharide (LPS) (1 μg/mL), f-met-leu-phe (fMLP) (100 nM), imiquimod (3 μg/mL), R848 (1.5 μg/mL), poly I:C (10 μg/mL), RV16 (multiplicity of infection (MOI)1), respiratory syncytial virus (RSV) (MOI1) or influenza virus (MOI1). Cell-free supernatants were collected after 1 h of neutrophil elastase (NE) and matrix metalloproteinase (MMP)-9 release, or after 24 h for CXCL8 release.. LPS, fMLP, imiquimod and R848 stimulated the release of CXCL8, NE and MMP-9 whereas poly I:C selectively induced CXCL8 release only. R848-induced CXCL8 release was enhanced in neutrophils from asthmatics compared with non-asthmatic cells (P < 0.01). RSV triggered the release of CXCL8 and NE from neutrophils, whereas RV16 or influenza had no effect.. Neutrophils release CXCL8, NE and MMP-9 in response to viral surrogates with R848-induced CXCL8 release being specifically enhanced in asthmatic neutrophils. Toll-like receptor (TLR7/8) dysregulation may play a role in neutrophilic inflammation in viral-induced exacerbations.

Topics: Adult; Asthma; Female; Humans; Inflammation; Interleukin-8; Leukocyte Elastase; Lipopolysaccharides; Male; Matrix Metalloproteinase 9; N-Formylmethionine Leucyl-Phenylalanine; Neutrophil Activation; Neutrophils; Orthomyxoviridae; Respiratory Syncytial Viruses; Symptom Flare Up; Toll-Like Receptors; Virus Diseases

Topics: Asthma; Comorbidity; Eosinophilia; Humans; Inflammation; Interleukin-8; Phenotype; Prevalence; Pulmonary Disease, Chronic Obstructive; Rhinitis; Syndrome

Neutrophilic inflammation has been implicated in non-eosinophilic asthma (NEA) in adults, but little is known about NEA in children/adolescents. We assessed clinical and inflammatory characteristics of NEA in adolescent asthma.. Airway inflammation, sputum endotoxin, airway hyper-reactivity, atopy and lung function were assessed in 77 adolescents with asthma and 68 without asthma (12-17 years). Asthma was identified on the basis of wheeze and asthma history.. The proportion of NEA (sputum eosinophils <2.5%) was 54%. In this group, atopy, sputum neutrophil, eosinophil, eosinophil cationic protein (ECP), endotoxin, neutrophil elastase and IL-8 levels were not different from those without asthma. In contrast, eosinophilic asthma (EA) was associated with atopy and sputum ECP and IL-8. The majority of NEA had no evidence of inflammation; only 14% had neutrophilia (≥61% neutrophils), compared with 11% of EA, and 15% of those without asthma. Small differences in FEV1 (NS) were found between EA and NEA, but symptom prevalence and severity was not different (63% of EA and 52% of NEA were classified moderate to severe).. NEA is common in adolescent asthma and has similar clinical characteristics as EA. Neutrophils do not appear to play a role in NEA in adolescents, and underlying mechanisms may not involve airway inflammation.

Topics: Adolescent; Asthma; Child; Eosinophils; Female; Humans; Inflammation; Interleukin-8; Leukocyte Count; Male; Neutrophils; Respiratory System; Sputum

Role of autophagy in neutrophil function and the association of autophagy and autophagy related (ATG) gene polymorphisms with asthma susceptibility were suggested. In this study, we investigated the genetic association of ATG5 and ATG7 polymorphisms with asthma risk, severity and neutrophilic airway inflammation.. We recruited 408 asthma patients and 201 healthy controls. Sputum neutrophil counts were determined by H&E staining. Serum interleukin 8 (IL-8) levels were measured by enzyme-linked immunosorbent assay (ELISA). Genetic polymorphisms of ATG5 (-769T>C, -335G>A, and 8830C>T) and ATG7 (-100A>G and 25108G>C) were genotyped. The functional activities of ATG5 -769T>C and -335G>A variants were investigated by luciferase reporter assays.. No associations of ATG5 and ATG7 polymorphisms with asthma susceptibility and severity were found. ATG5 -769T>C and -335G>A were in complete linkage disequilibrium. In the asthma group, GA/AA genotypes at ATG5 -335G>A were associated with higher neutrophil counts in sputum (p < 0.05); CC/TT genotype at ATG5 8830C>T associated with lower FEV1% predicted value (p < 0.05). DNA fragments containing ATG5 -769T and -335G alleles had higher promoter activities compared to those with -769C and -335A in both human airway epithelial cells (A549, p < 0.01) and human mast cell (HMC-1, p < 0.001). GG and CC genotype at ATG7 -100A>G and 25108G>C were significantly associated with high serum levels of IL-8 (p < 0.05 for both variants).. Genetic polymorphisms of ATG5 and ATG7 could contribute to neutrophilic airway inflammation in the pathogenesis of adult asthma.

Topics: Adolescent; Adult; Asthma; Autophagy; Autophagy-Related Protein 5; Autophagy-Related Protein 7; Case-Control Studies; Cell Line; Female; Gene Frequency; Genes, Reporter; Genetic Predisposition to Disease; Haplotypes; Heterozygote; Homozygote; Humans; Interleukin-8; Male; Middle Aged; Neutrophil Infiltration; Neutrophils; Phenotype; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Risk Factors; Severity of Illness Index; Transfection; Young Adult

Several cytokines may play roles in the immunological pathogenesis of mycoplasmal pneumonia caused by Mycoplasma pneumoniae. In this study, we investigated serum cytokine profiles in children with mycoplasmal pneumonia. The serum levels of interleukin (IL)-8, IL-10, and IL-18 were examined using ELISA kits in 34 patients with M. pneumoniae infection (Group 1, 11 with severe mycoplasmal pneumonia; Group 2, 13 with mild mycoplasmal pneumonia; Group 3, 10 with asthma) and 32 age-matched, non-infected controls. The serum levels of IL-8, IL-10, and IL-18 increased significantly in patients with mycoplasmal pneumonia compared with those in controls (P<0.01). The serum levels of IL-10 decreased significantly in Group 1 compared with those in Group 2 (P<0.01). The serum levels of IL-18 increased significantly in Group 1 compared with those in Group 2 (P<0.01). The serum levels of IL-10 and IL-18 decreased significantly in 10 M. pneumoniae-infected patients with asthma compared with those in 24 M. pneumoniae-infected patients without asthma (P<0.01). We examined the level of interleukins (IL-8, IL-10 and IL-18) after the patients started therapy. The data showed that IL-18 were lower after therapy (P<0.01). Collectively, our data suggested that these cytokines may be involved in the pathogenesis of mycoplasmal pneumonia.

Topics: Anti-Bacterial Agents; Asthma; C-Reactive Protein; Case-Control Studies; Child; Child, Preschool; Cytokines; Enzyme-Linked Immunosorbent Assay; Erythrocyte Count; Female; Humans; Infant; Interleukin-10; Interleukin-8; Lymphocyte Count; Male; Mycoplasma pneumoniae; Pneumonia, Mycoplasma

Glucocorticoids are the most effective anti-inflammatory agent in treating pulmonary diseases typically accompanied by hypoxia. Our previous study has demonstrated that glucocorticoid receptor α (GRα) expression is reduced in hypoxia but the underlying mechanism remains elusive. In this study we aim to identify the signaling pathway involved in hypoxia-induced down-regulation of GRα, and whether hypoxia affects nuclear translocation of GRα.. Female C57BL/6 mice were sensitized with saline or ovalbumin (OVA) as the in vivo model. Mice were divided into control and OVA groups, and their lung histology and the expression of hypoxia inducible factor (HIF-1) and GRα were examined. A549 cells were exposed to chemical hypoxia as the in vitro model, where mitogen-activated protein kinases (MAPKs) were inhibited specifically by SB203580. Next, under normal or hypoxic conditions, the expression of GRα, MAPKs and HIF-1 signal protein were determined by Western blot analysis, and GRα translocation were observed through live-cell imaging.. In OVA challenged mice the expression of GRα was down-regulated whereas HIF-1 was up-regulated. Hypoxia caused a time-dependent decrease of GRα expression, and activated multiple signaling pathways including MAPKs and HIF-1. Moreover, GRα expression increased with MAPK inhibition. Interestingly, only MAPK inhibitor SB203580, but not JNK inhibitor SP600125 or ERK inhibitor U0126 improved the expression of GRα under hypoxic condition. GRα nuclear translocation was also significantly inhibited by hypoxia.. Hypoxia down-regulated the expression of GRα through p38 signaling pathway, as well as inhibited GRα nuclear translocation significantly.

Topics: Animals; Asthma; Down-Regulation; Female; Hypoxia; Hypoxia-Inducible Factor 1, alpha Subunit; Interleukin-8; Janus Kinases; Lung; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinases; Ovalbumin; Protein Kinase Inhibitors; Protein Transport; Receptors, Glucocorticoid; Transfection

Asthma is a heterogeneous chronic inflammatory disease in which host defense against respiratory viruses such as human rhinovirus (HRV) may be abnormal. This is a matter of some controversy, with some investigators reporting reduced type I interferon (IFN) synthesis and others suggesting that type I IFN synthesis is relatively normal in asthma.. The objective of this study was to examine the responsiveness of circulating mononuclear cells to HRV in a large cohort of participants with poorly controlled asthma and determine whether IFN-α and IFN-β synthesis varies across different inflammatory phenotypes.. Eligible adults with asthma (n = 86) underwent clinical assessment, sputum induction, and blood sampling. Asthma inflammatory subtypes were defined by sputum cell count, and supernatant assessed for IL-1β. Peripheral blood mononuclear cells (PBMCs) were exposed to HRV serotype 1b, and IFN-α and IFN-β release was measured by enzyme-linked immunosorbent assay.. Participants (mean age, 59 years; atopy, 76%) had suboptimal asthma control (mean asthma control questionnaire 6, 1.7). In those with neutrophilic asthma (n = 12), HRV1b-stimulated PBMCs produced significantly less IFN-α than PBMCs from participants with eosinophilic (n = 35) and paucigranulocytic asthma (n = 35). Sputum neutrophil proportion and the dose of inhaled corticosteroids were independent predictors of reduced IFN-α production after HRV1b exposure.. Antiviral type I IFN production is impaired in those with neutrophilic airway inflammation and in those prescribed high doses of inhaled corticosteroids. Our study is an important step toward identifying those with poorly controlled asthma who might respond best to inhaled IFN therapy during exacerbations.

Topics: Administration, Inhalation; Adrenal Cortex Hormones; Adult; Aged; Aged, 80 and over; Asthma; Chemokine CXCL10; Cross-Sectional Studies; Enzyme-Linked Immunosorbent Assay; Female; Forced Expiratory Volume; Glucocorticoids; HeLa Cells; Humans; Interferon-alpha; Interferon-beta; Interleukin-1beta; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Neutrophils; Rhinovirus; Severity of Illness Index; Sputum; Vital Capacity; Young Adult

Obstructive sleep apnoea (OSA) is frequently observed in severe asthma but the causal link between the 2 diseases remains hypothetical. The role of OSA-related systemic and airway neutrophilic inflammation in asthma bronchial inflammation or remodelling has been rarely investigated. The aim of this study was to compare hallmarks of inflammation in induced sputum and features of airway remodelling in bronchial biopsies from adult patients with severe asthma with and without OSA.. An overnight polygraphy was performed in 55 patients referred for difficult-to-treat asthma, who complained of nocturnal respiratory symptoms, poor sleep quality or fatigue. We compared sputum analysis, reticular basement membrane (RBM) thickness, smooth muscle area, vascular density and inflammatory cell infiltration in bronchial biopsies.. In total, 27/55 patients (49%) had OSA diagnosed by overnight polygraphy. Despite a moderate increase in apnoea-hypopnoea index (AHI; 14.2 ± 1.6 event/h [5-35]), the proportion of sputum neutrophils was higher and that of macrophages lower in OSA than non-OSA patients, with higher levels of interleukin 8 and matrix metalloproteinase 9. The RBM was significantly thinner in OSA than non-OSA patients (5.8 ± 0.4 vs. 7.8 ± 0.4 μm, p<0.05). RBM thickness and OSA severity assessed by the AHI were negatively correlated (rho = -0.65, p<0.05). OSA and non-OSA patients did not differ in age, sex, BMI, lung function, asthma control findings or treatment.. Mild OSA in patients with severe asthma is associated with increased proportion of neutrophils in sputum and changes in airway remodelling.

Topics: Adolescent; Adult; Aged; Asthma; Case-Control Studies; Female; Humans; Inflammation; Interleukin-8; Macrophages; Male; Matrix Metalloproteinase 9; Middle Aged; Neutrophils; Prospective Studies; Respiratory System; Sleep Apnea, Obstructive; Sputum; Young Adult

To study the role of serum neutrophil elastase (NE) level in acute exacerbation of asthma in preschool children.. A total of 85 preschool children who were diagnosed with asthma between January 2008 and January 2010 were classified into acute exacerbation group (n=44) and non-acute exacerbation group (n=41). Thirty-five children who received physical examination served as the control group. The enzyme-linked immunosorbent assay was used to determine the serum levels of NE and interleukin-8 (IL-8). The receiver operating characteristic (ROC) curve was used for NE evaluation.. Both the acute and non-acute exacerbation groups had higher serum levels of NE and IL-8 than the control group, and the acute exacerbation group had significantly higher serum levels of NE and IL-8 than the non-acute exacerbation group (P<0.05). The serum level of NE was positively correlated with that of IL-8 (r=0.48, P<0.05). With serum NE level >27.73 μg/L as the cut-off value for diagnosing acute exacerbation of asthma, the sensitivity was 65.9%, the specificity was 95.1%, and the area under the ROC curve was 0.87 (P<0.01).. The determination of serum NE level in preschool children with asthma helps to diagnose the acute exacerbation of asthma.

Topics: Asthma; Child; Child, Preschool; Female; Humans; Interleukin-8; Leukocyte Elastase; Male; ROC Curve

Although modern treatment of asthma improves asthma control, some patients still experience exacerbations. The aim of the present study was to detect predictors of asthmatic exacerbations Methods: We included patients with asthma followed up in asthma clinics of 2 tertiary University hospitals. Demographic and functional characteristics, levels of exhaled NO, and inflammatory biomarkers (IL-13, ΕCP και IL-8) and cell counts in induced sputum were recorded at baseline. Measurements were performed with the patients in stability and were considered as their personal best. Patients received optimal treatment with good compliance and were followed up for 1 year for asthma exacerbations occurrence. Evaluation of the effect of recorded parameters on asthma exacerbations was performed with univariate and multivariate Poisson regression analysis.. 171 patients (118 female) with bronchial asthma (mean age 51.6 ± 13.2 years) were included in the study. The mean number of exacerbations in 1 year of follow up was 0.4 ± 0.8 while the majority of patients (71.9%) did not experience any exacerbation. In multivariate Poisson Regression analysis only 3 characteristics were predictors of future exacerbations: FEV1 [IRR(95% CI)], [0.970(0.954-0.987)], p = 0.001, high BMI [1.078(1.030-1.129)], p = 0.001, and the need for permanent treatment with oral corticosteroids for asthma control maintenance [2.542(1.083-5.964)], p = 0.032 CONCLUSION: Optimal guideline-based asthma management results in minimal occurrence of exacerbations in the majority of patients. Predictors of exacerbations are low FEV1 levels in stability, high BMI and the need for permanent treatment with oral corticosteroids.

Topics: Adult; Anti-Asthmatic Agents; Asthma; Biomarkers; Disease Progression; Eosinophil Cationic Protein; Female; Follow-Up Studies; Glucocorticoids; Greece; Humans; Interleukin-13; Interleukin-8; Male; Middle Aged; Respiratory Function Tests; Risk Assessment; Sputum; Symptom Flare Up

Exacerbations of asthma and COPD are triggered by rhinoviruses. Uncontrolled inflammatory pathways, pathogenic bacterial burden and impaired antiviral immunity are thought to be important factors in disease severity and duration. Macrolides including azithromycin are often used to treat the above diseases, but exhibit variable levels of efficacy. Inhaled corticosteroids are also readily used in treatment, but may lack specificity. Ideally, new treatment alternatives should suppress unwanted inflammation, but spare beneficial antiviral immunity.. In the present study, we screened 225 novel macrolides and tested them for enhanced antiviral activity against rhinovirus, as well as anti-inflammatory activity and activity against Gram-positive and Gram-negative bacteria. Primary bronchial epithelial cells were grown from 10 asthmatic individuals and the effects of macrolides on rhinovirus replication were also examined. Another 30 structurally similar macrolides were also examined.. The oleandomycin derivative Mac5, compared with azithromycin, showed superior induction (up to 5-fold, EC50 = 5-11 μM) of rhinovirus-induced type I IFNβ, type III IFNλ1 and type III IFNλ2/3 mRNA and the IFN-stimulated genes viperin and MxA, yet had no effect on IL-6 and IL-8 mRNA. Mac5 also suppressed rhinovirus replication at 48 h, proving antiviral activity. Mac5 showed antibacterial activity against Gram-positive Streptococcus pneumoniae; however, it did not have any antibacterial properties compared with azithromycin when used against Gram-negative Escherichia coli (as a model organism) and also the respiratory pathogens Pseudomonas aeruginosa and non-typeable Haemophilus influenzae. Further non-toxic Mac5 derivatives were identified with various anti-inflammatory, antiviral and antibacterial activities.. The data support the idea that macrolides have antiviral properties through a mechanism that is yet to be ascertained. We also provide evidence that macrolides can be developed with anti-inflammatory, antibacterial and antiviral activity and show surprising versatility depending on the clinical need.

Topics: Anti-Bacterial Agents; Anti-Inflammatory Agents, Non-Steroidal; Antiviral Agents; Asthma; Bronchi; Cells, Cultured; Drug Discovery; Drug Evaluation, Preclinical; Epithelial Cells; Escherichia coli; Gram-Negative Bacteria; Gram-Positive Bacteria; Haemophilus influenzae; Humans; Interferon-beta; Interferons; Interleukin-6; Interleukin-8; Macrolides; Myxovirus Resistance Proteins; Oxidoreductases Acting on CH-CH Group Donors; Proteins; Pseudomonas aeruginosa; Rhinovirus; Virus Replication

To study the effects of the change in transient receptor potential vanilloid 1 (TRPV1) channel activity on the degree of airway inflammation in asthmatic mice.. BALB/c mice were randomly divided into control, asthma, capsaicin (TRPV1 agonist), capsazepine (TRPV1 antagonist), and dexamethasone groups. The asthmatic mouse model was established by intraperitoneal injection of mixed ovalbumin-aluminium hydroxide solution and ultrasonic atomization with OVA for sensitization and challenge. The capsaicin, capsazepine, and dexamethasone groups were given intraperitoneal injection of capsaicin (30 μg/kg), capsazepine (10 μmol/kg), and dexamethasone (2 mg/kg) respectively, at 30 minutes before challenge. Hematoxylin and eosin staining was used to observe the degree of pulmonary inflammation. ELISA was used to measure the content of interleukin-8 (IL-8) and interleukin-13 (IL-13) in bronchoalveolar lavage fluid (BALF). Real-Time PCR was used to measure the relative content of TRPV1 mRNA in lung tissue.. Compared with the asthma group, the capsazepine and dexamethasone groups showed reduced pulmonary inflammation, while the capsaicin group showed aggravated pulmonary inflammation. Compared with the control group, the asthma and capsaicin groups showed increases in the content of IL-13 and IL-8 in BALF and the mRNA expression of TRPV1 in lung tissue (P<0.05). Compared with the asthma group, the capsazepine and dexamethasone groups showed reductions in the content of IL-13 and IL-8 in BALF and the mRNA expression of TRPV1 in lung tissue (P<0.05). The capsaicin group showed increases in the content of IL-13 and IL-8 in BALF (P<0.05).. TRPV1 channel agonist and antagonist can influence the degree of airway inflammation in asthmatic mice. Dexamethasone may reduce airway inflammation through regulating TRPV1 level.

Topics: Animals; Asthma; Female; Interleukin-13; Interleukin-8; Lung; Mice; Mice, Inbred BALB C; RNA, Messenger; TRPV Cation Channels

Bronchial asthma (BA) is a chronic airway disease characterized by airway hyperresponsiveness and remodeling, which are intimately linked to chronic airway inflammation. Reactive oxygen species (ROS) such as hydrogen peroxide are generated by inflammatory cells that are involved in the pathogenesis of BA. However, the role of ROS in the management of BA patients is not yet clear. We attempted to determine the role of ROS as a biomarker in the clinical setting of BA.. We enrolled patients with BA from 2013 through 2015 and studied the degrees of asthma control, anti-asthma treatment, pulmonary function test results, fractional exhaled nitric oxide (FeNO), serum reactive oxygen metabolite (ROM) levels, and serum levels of interleukin (IL)-6 and IL-8.. We recruited 110 patients with BA. Serum ROM levels correlated with white blood cell (WBC) count (rs = 0.273, p = 0.004), neutrophil count (rs = 0.235, p = 0.014), CRP (rs = 0.403, p < 0.001), and IL-6 (rs = 0.339, p < 0.001). Serum ROM levels and IL-8 and CRP levels negatively correlated with %FEV1 (rs = -0.240, p = 0.012, rs = -0.362, p < 0.001, rs = -0.197, p = 0.039, respectively). Serum ROM levels were significantly higher in patients who experienced severe exacerbation within 3 months than in patients who did not (339 [302-381] vs. 376 [352-414] CARR U, p < 0.025). Receiver-operating characteristics analysis showed that ROM levels correlated significantly with the occurrence of severe exacerbation (area under the curve: 0.699, 95% CI: 0.597-0.801, p = 0.025).. Serum levels of ROM were significantly associated with the degrees of airway obstruction, WBC counts, neutrophil counts, IL-6, and severe exacerbations. This biomarker may be useful in predicting severe exacerbations of BA.

Topics: Adult; Anti-Asthmatic Agents; Asthma; Biomarkers; Breath Tests; Female; Humans; Interleukin-6; Interleukin-8; Leukocyte Count; Male; Middle Aged; Reactive Oxygen Species; Respiratory Function Tests; ROC Curve

Steroid resistant (SR) asthma is characterized by persistent airway inflammation that fails to resolve despite treatment with high doses of corticosteroids. Furthermore, SR patient airways show increased numbers neutrophils, which are less responsive to glucocorticoid. The present study seeks to determine whether dexamethasone (DEX) has different effect on neutrophils from steroid sensitive (SS) asthmatics compared to SR asthmatics.. Adults with asthma (n = 38) were classified as SR or SS based on changes in lung FEV1% following a one-month inhaled corticosteroid (ICS) treatment. Blood samples were collected from all patients during their first visit of the study. Neutrophils isolated from the blood were cultured with dexamethasone and/or atopic asthmatic serum for 18 h. The mRNA expression of mitogen-activated protein kinase phosphatase-1 (MKP-1), a glucocorticoid transactivation target, and glucocorticoid-induced transcript 1 (GLCCI1), an early marker of glucocorticoid-induced apoptosis whose expression was associated with the response to inhaled glucocorticoids in asthma , was determined by real-time PCR, and ELISA was used to assess the pro-inflammatory cytokine IL-8 levels in the supernatant. Constitutive neutrophil apoptosis was detected by flow cytometry.. DEX exerted different effects on neutrophils from patients with SS asthma and SR asthma, which may contribute to glucocorticoid insensitivity.

Topics: Administration, Inhalation; Adult; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Apoptosis; Asthma; Cells, Cultured; Dexamethasone; Dose-Response Relationship, Drug; Drug Resistance; Dual Specificity Phosphatase 1; Forced Expiratory Volume; Glucocorticoids; Humans; Interleukin-8; Lung; Male; Middle Aged; Neutrophils; Receptors, Glucocorticoid; Time Factors; Treatment Outcome

Topics: Anthracenes; Asthma; Cell Line, Tumor; Drug Synergism; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-6; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Lipopolysaccharides; MAP Kinase Signaling System; Peptides; Phosphorylation

Airway inflammation is involved in the pathogenesis of bronchopulmonary dysplasia (BPD). The aim of the study was to evaluate the inflammatory activity in plasma and exhaled air in very low birth weight (VLBW) BPD survivors at school age.. Twenty-one 6-14-year-old former VLBW (birth weight ≤1,500 g) children with severe radiographic BPD (radBPD), 19 without radBPD (nonBPD group) and 19 non-asthmatic term controls underwent measurement of eosinophil cationic protein, IL-6, IL-8, adiponectin, adipsin, leptin, and resistin in plasma, leukotriene B4 and 8-isoprostane in exhaled breath condensate, and NO in exhaled breath. Background data were obtained from patient records, clinical examination and parental questionnaire. Both univariate and multivariate models were applied in the statistical analysis.. There were no significant differences between the groups in any of the inflammatory markers measured. Five (25%) radBPD and 2 (11%) nonBPD children reported asthma (P = 0.058). In logistic regression analysis, exposure to chorioamnionitis was associated with low IL-8 (OR 29.0, 95% CI 3.27-258) and postnatal corticosteroid therapy with high adiponectin (OR 32.0, 95% CI 1.29-793). High body mass index standard deviation score (BMI-SDS) was associated with high plasma adipsin (OR 2.47, 95% CI 1.07-5.75) and leptin (OR 5.76, 95%CI 1.83-18.2) levels.. The inflammatory activity seems to decrease by school age in VLBW BPD survivors. Chorioamnionitis and postnatal corticosteroid treatment may modulate the inflammatory responsiveness in VLBW subjects even up to school age. The respiratory outcome in VLBW infants might be improved by preventing excessive weight gain.

Topics: Adiponectin; Adolescent; Asthma; Biomarkers; Body Mass Index; Bronchopulmonary Dysplasia; Child; Chorioamnionitis; Complement Factor D; Female; Finland; Glucocorticoids; Humans; Infant, Newborn; Infant, Very Low Birth Weight; Interleukin-6; Interleukin-8; Leptin; Logistic Models; Pregnancy; Survivors

Elite athletes frequently experience asthma and airway hyperresponsiveness (AHR). We aimed to investigate predictors of airway pathophysiology in a group of unselected elite summer-sport athletes, training for the summer 2008 Olympic Games, including markers of airway inflammation, systemic inflammation, and training intensity.. Fifty-seven Danish elite summer-sport athletes with and without asthma symptoms all gave a blood sample for measurements of high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor alpha (TNF-α), completed a respiratory questionnaire, and underwent spirometry. Bronchial challenges with mannitol were performed in all 57 athletes, and 47 agreed to perform an additional methacholine provocation.. Based on a physician's diagnosis, 18 (32%) athletes were concluded to be asthmatic. Asthmatic subjects trained more hours per week than the 39 nonasthmatics (median (min-max): 25 h·wk (14-30) versus 20 h·wk (11-30), P = 0.001). AHR to both methacholine and mannitol (dose response slope) increased with the number of weekly training h (r = 0.43, P = 0.003, and r = 0.28, P = 0.034, respectively). Serum levels of IL-6, IL-8, TNF-α, and hs-CRP were similar between asthmatics and nonasthmatics. However, there was a positive association between the degree of AHR to methacholine and serum levels of TNF-α (r = 0.36, P = 0.04). Fifteen out of 18 asthmatic athletes were challenged with both agents. In these subjects, no association was found between the levels of AHR to mannitol and methacholine (r = 0.032, P = 0.91).. AHR in elite athletes is related to the amount of weekly training and the level of serum TNF-α. No association was found between the level of AHR to mannitol and methacholine in the asthmatic athletes.

Topics: Adult; Asthma; Biomarkers; Bronchial Hyperreactivity; Bronchial Provocation Tests; C-Reactive Protein; Cross-Sectional Studies; Female; Humans; Interleukin-6; Interleukin-8; Male; Mannitol; Methacholine Chloride; Physical Education and Training; Spirometry; Sports; Tumor Necrosis Factor-alpha; Young Adult

Topics: Asthma; Body Mass Index; Cell Count; Eosinophils; Forced Expiratory Volume; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Neutrophils; Obesity; Ozone; Retrospective Studies; Spirometry; Sputum; Vital Capacity

Children and adults with asthma and impaired lung function have been reported to have low-grade systemic inflammation, but it is unknown whether this inflammation starts before symptoms and in particular whether low-grade inflammation is present in asymptomatic neonates with reduced lung function.. We sought to investigate the possible association between neonatal lung function and biomarkers of systemic inflammation.. Plasma levels of high-sensitivity C-reactive protein (hs-CRP), IL-1β, IL-6, TNF-α, and CXCL8 (IL-8) were measured at age 6 months in 300 children of the Copenhagen Prospective Study on Asthma in Childhood2000 birth cohort who had completed neonatal lung function testing at age 4 weeks. Associations between neonatal lung function indices and inflammatory biomarkers were investigated by conventional statistics and unsupervised principal component analysis.. The neonatal forced expiratory volume at 0.5 seconds was inversely associated with hs-CRP (β-coefficient, -0.12; 95% CI, -0.21 to -0.04; P < .01) and IL-6 (β-coefficient, -0.10; 95% CI, -0.18 to -0.01; P = .03) levels. The multivariate principal component analysis approach, including hs-CRP, IL-6, TNF-α, and CXCL8, confirmed a uniform upregulated inflammatory profile in children with reduced forced expiratory volume at 0.5 seconds (P = .02). Adjusting for body mass index at birth, maternal smoking, older children in the home, neonatal bacterial airway colonization, infections 14 days before, and asthmatic symptoms, as well as virus-induced wheezing, at any time before biomarker assessment at age 6 months did not affect the associations.. Diminished neonatal lung function is associated with upregulated systemic inflammatory markers, such as hs-CRP.

Topics: Adult; Asthma; Biomarkers; Birth Weight; Body Mass Index; C-Reactive Protein; Denmark; Female; Forced Expiratory Volume; Humans; Infant; Infant, Newborn; Inflammation; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Maternal Exposure; Multivariate Analysis; Principal Component Analysis; Prospective Studies; Smoking; Tumor Necrosis Factor-alpha

Adenosine activated mast cells have been long implicated in allergic asthma and studies in rodent mast cells have assigned the A3 adenosine receptor (A3R) a primary role in mediating adenosine responses. Here we analyzed the functional impact of A3R activation on genes that are implicated in tissue remodeling in severe asthma in the human mast cell line HMC-1 that shares similarities with lung derived human mast cells. Quantitative real time PCR demonstrated upregulation of IL6, IL8, VEGF, amphiregulin and osteopontin. Moreover, further upregulation of these genes was noted upon the addition of dexamethasone. Unexpectedly, activated A3R down regulated its own expression and knockdown of the receptor replicated the pattern of agonist induced gene upregulation. This study therefore identifies the human mast cell A3R as regulator of tissue remodeling gene expression in human mast cells and demonstrates a heretofore-unrecognized mode of feedback regulation that is exerted by this receptor.

Topics: Adenosine; Adenosine A3 Receptor Antagonists; Amphiregulin; Anti-Inflammatory Agents; Asthma; Cell Line, Tumor; Dexamethasone; Down-Regulation; EGF Family of Proteins; Humans; Interleukin-6; Interleukin-8; Jurkat Cells; Lung; Mast Cells; Neovascularization, Pathologic; Osteopontin; Phosphorylation; Receptor, Adenosine A3; RNA Interference; RNA, Small Interfering; Up-Regulation; Vascular Endothelial Growth Factor A

Asthma is characterized by airway inflammation and remodeling and CXCL8 is a CXC chemokine that drives steroid-resistant neutrophilic airway inflammation. We have shown that airway smooth muscle (ASM) cells isolated from asthmatic individuals secrete more CXCL8 than cells from nonasthmatic individuals. Here we investigated chromatin modifications at the CXCL8 promoter in ASM cells from nonasthmatic and asthmatic donors to further understand how CXCL8 is dysregulated in asthma. ASM cells from asthmatic donors had increased histone H3 acetylation, specifically histone H3K18 acetylation, and increased binding of histone acetyltransferase p300 compared with nonasthmatic donors but no differences in CXCL8 DNA methylation. The acetylation reader proteins Brd3 and Brd4 were bound to the CXCL8 promoter and Brd inhibitors inhibited CXCL8 secretion from ASM cells by disrupting Brd4 and RNA polymerase II binding to the CXCL8 promoter. Our results show a novel dysregulation of CXCL8 transcriptional regulation in asthma characterized by a promoter complex that is abnormal in ASM cells isolated from asthmatic donors and can be modulated by Brd inhibitors. Brd inhibitors may provide a new therapeutic strategy for steroid-resistant inflammation.

Topics: Acetylation; Airway Remodeling; Asthma; CCAAT-Enhancer-Binding Protein-beta; Cell Cycle Proteins; Cells, Cultured; DNA Methylation; DNA-Binding Proteins; Histones; Humans; Inflammation; Interleukin-8; Muscle, Smooth; Myocytes, Smooth Muscle; Nerve Tissue Proteins; Nuclear Proteins; p300-CBP Transcription Factors; Primary Cell Culture; Promoter Regions, Genetic; Protein Binding; RNA Polymerase II; RNA-Binding Proteins; Transcription Factor RelA; Transcription Factors; Transcription, Genetic

Previous studies investigating the role of toll-like receptors (TLRs) in asthma have been inconclusive. It has remained elusive whether the toll-like receptors (TLR2)/mycloid differentiation factor 88 (MyD88)/nuclear factor (NF)-κB signaling pathway is involved in lipoxin A4 (LXA4)-induced protection against asthma. Therefore, the present study investigated whether ovalbumin (OVA)-induced airway inflammation is mediated by upregulation of the TLR2/MyD88/NF-κB signaling pathway, and whether it proceeds via the inhibition of the activation of the LXA4 receptor and anti-interleukin (IL)-1β antibodies. Mice with airway inflammation induced by OVA administration were treated with or without a LXA4 receptor agonist, BML-111 and anti-IL-1β antibody. Serum levels of IL-1β, IL-4, IL-8 and interferon-γ (IFN-γ) were assessed, and levels of IL-1β, IL-4, IL-8 and OVA-immunoglobulin (Ig)E, as well as leukocyte counts in the bronchoalveolar lavage fluid (BALF) were measured. Pathological features and expression of TLR2, MyD88 and NF-κB in the lungs were analyzed. Expression of TLR2 and MyD88, and activation of NF-κB in leukocytes as well as levels of IL-4, IL-6 and IL-8 released from leukocytes exposed to IL-1β were assessed. OVA treatment increased the levels of IL-1β, IL-4 and IL-8 in the serum and BLAF, the number of leukocytes and the levels of OVA-IgE in the BALF, the expression of TLR2 and MyD88, and the activation of NF-κB in the lung. These increments induced by OVA were inhibited by treatment with BML-111 and anti-IL-1β antibodies. Treatment of the leukocytes with BML-111 or TLR2 antibody, or MyD88 or NF-κB inhibitor, all blocked the IL-1β-triggered production of IL-4, IL-6 and IL-8 and activation of NF-κB. Treatment of the leukocytes with BML-111 or TLR2 antibody suppressed IL-1β-induced TLR2 and MyD88 expression. The present study therefore suggested that OVA-induced airway inflammation is mediated by the TLR2/MyD88/NF-κB pathway. IL-1β has a pivotal role in the airway inflammation and upregulation of the TLR2/MyD88/NF-κB pathway induced by OVA. BML-111 and anti-IL-1β antibody restrains the OVA-induced airway inflammation via downregulation of the TLR2/MyD88/NF-κB pathway.

Topics: Animals; Anti-Asthmatic Agents; Antibodies, Monoclonal; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Gene Expression Regulation; Heptanoic Acids; Interleukin-1beta; Interleukin-4; Interleukin-6; Interleukin-8; Lipoxins; Male; Mice; Mice, Inbred BALB C; Myeloid Differentiation Factor 88; NF-kappa B; Ovalbumin; Receptors, Formyl Peptide; Signal Transduction; Toll-Like Receptor 2

The role of mitogen-activated protein kinases (MAPK) in regulating the inflammatory response in the airways of patients with chronic obstructive pulmonary disease (COPD) and asthmatic patients is unclear.. To investigate the expression of activated MAPK in lungs of COPD patients and in bronchial biopsies of asthmatic patients and to study MAPK expression in bronchial epithelial cells in response to oxidative and inflammatory stimuli.. Immunohistochemical expression of phospho (p)-p38 MAPK, p-JNK1 and p-ERK1/2 was measured in bronchial mucosa in patients with mild/moderate (n = 17), severe/very severe (n = 16) stable COPD, control smokers (n = 16), control non-smokers (n = 9), in mild asthma (n = 9) and in peripheral airways from COPD patients (n = 15) and control smokers (n = 15). Interleukin (IL)-8 and MAPK mRNA was measured in stimulated 16HBE cells.. No significant differences in p-p38 MAPK, p-JNK or p-ERK1/2 expression were seen in bronchial biopsies and peripheral airways between COPD and control subjects. Asthmatics showed increased submucosal p-p38 MAPK expression compared to COPD patients (p < 0.003) and control non-smokers (p < 0.05). Hydrogen peroxide (H₂O₂), cytomix (tumour necrosis factor-α + IL-1β + interferon-γ) and lipopolysaccharide (LPS) upregulated IL-8 mRNA at 1 or 2 h. p38 MAPKα mRNA was significantly increased after H₂O₂ and LPS treatment. JNK1 and ERK1 mRNA were unchanged after H₂O₂, cytomix or LPS treatments.. p-p38 MAPK expression is similar in stable COPD and control subjects but increased in the bronchi of mild asthmatics compared to stable COPD patients. p38 MAPK mRNA is increased after bronchial epithelial challenges in vitro. These data together suggest a potential role for this MAPK in Th2 inflammation and possibly during COPD exacerbations.

Topics: Aged; Asthma; Blotting, Western; Bronchi; Case-Control Studies; Cell Line; Female; Humans; Immunohistochemistry; Interleukin-8; Lymphocyte Count; Male; Middle Aged; p38 Mitogen-Activated Protein Kinases; Pulmonary Disease, Chronic Obstructive; Respiratory Mucosa; Transcription Factor RelA

Airway epithelial cells (AEC) are key contributors to immune function in the lungs but little is known about their role and function in children.. Having previously established that nasal AEC mediator release correlates with that of bronchial AEC, we assessed AEC responses in children with and without a history of wheeze.. Nasal AEC cultures were established from children (0.6-14.9 years) undergoing elective surgical procedures under general anaesthetic categorised as atopic asthmatic (n = 12), virus-induced wheeze (n = 8) or children without wheeze (n = 32). Mediator release by AEC monolayers at passage 2 was determined by cytometric bead array assay or ELISA.. Unstimulated AEC from children with a history of wheeze produced significantly less IL-8, IL-6, MCP-1 and G-CSF than AEC from healthy controls. There were no group differences in AEC release of VEGF, RANTES, MMP-9 or TIMP-1. After stimulation with the pro-inflammatory cytokines IL-1β and TNFα, AEC from children with current wheeze produced significantly less IL-8, IL-6 and MCP-1 than children without wheeze. Release of G-CSF, VEGF, MMP-9 and TIMP-1 did not differ between the wheeze and control group. There were no differences in mediator release between subjects with atopic asthma and those with virus-induced wheeze or between atopic and non-atopic controls. On multivariate analysis, wheeze was the only significant predictor of AEC mediator release.. Intrinsic differences in AEC from children with a history of wheeze may reflect a defect in cytokine production in vivo or an altered state of differentiation in vitro, independent of atopic status.

Topics: Adolescent; Asthma; Chemokine CCL2; Chemokine CCL5; Child; Child, Preschool; Cytokines; Female; Granulocyte Colony-Stimulating Factor; Humans; Infant; Inflammation Mediators; Interleukin-6; Interleukin-8; Male; Respiratory Mucosa; Respiratory Sounds; Tissue Inhibitor of Metalloproteinase-1; Vascular Endothelial Growth Factor A

Asthma is a heterogeneous disease, and a great majority of pediatric patients with asthma demonstrate atopic characteristics and develop a Th2 type cytokine response. Nonatopic asthma, on the other hand, is seen more rarely.. In this study, levels of IL-5, IL-8 and MMP-9 were measured in exhaled breath condensate (EBC) of the subjects to demonstrate the extent of tissue damage as well as eosinophilic and neutrophilic inflammation in children with atopic and nonatopic asthma. A total of 37 children with atopic asthma and 37 children with nonatopic asthma were enrolled in the study. Patients who exhibited protease positive aeroallergen (House dust mite, mould mix, olea, grass mix) sensitivity in allergen skin prick test were included in the atopic asthma group. To evaluate the EBC, the fluid content of the breath was collected by having the patients exhale into an EBC device, after which the IL-5, IL-8 and MMP-9 levels were assayed using the ELISA method.. The atopic asthmatics exhibited significantly higher IL-5 levels in their EBC samples than the nonatopic asthmatics (0.271 [0.198-0.489] pg/ml and 0.198 [0.125-0.344] pg/ml, respectively, p = 0.04), while no significant differences were observed in the levels of IL-8 and MMP-9 in the EBC samples of the atopic and nonatopic asthmatics.. IL-5 levels, as a marker of eosinophilic inflammation, were demonstrated to be higher in the children with atopic asthma when compared to those with nonatopic asthma in EBC. The fact that no significant difference was apparent in the IL-8 levels between the groups suggests that it is the severity of the disease rather than the atopic state that plays an important role in IL-8 levels. Since no difference was recorded between the groups in terms of MMP-9 levels, lung damage in asthma sufferers seems to develop independent of atopia.

Topics: Asthma; Biomarkers; Breath Tests; Case-Control Studies; Child; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-5; Interleukin-8; Male; Matrix Metalloproteinase 9; Th2 Cells; Turkey

We sought to identify cells and cytokines in bronchoalveolar lavage (BAL) fluids that distinguish asthma from healthy control subjects and those that distinguish controlled asthma from uncontrolled asthma. Following informed consent, 36 human subjects were recruited for this study. These included 11 healthy control subjects, 15 subjects with controlled asthma with FEV1≥80% predicted and 10 subjects with uncontrolled asthma with FEV1 <80% predicted. BAL fluid was obtained from all subjects. The numbers of different cell types and the levels of 48 cytokines were measured in these fluids. Compared to healthy control subjects, patients with asthma had significantly more percentages of eosinophils and neutrophils, IL-1RA, IL-1α, IL-1β, IL-2Rα, IL-5, IL-6, IL-7, IL-8, G-CSF, GROα (CXCL1), MIP-1β (CCL4), MIG (CXCL9), RANTES (CCL5) and TRAIL in their BAL fluids. The only inflammatory markers that distinguished controlled asthma from uncontrolled asthma were neutrophil percentage and IL-8 levels, and both were inversely correlated with FEV1. We examined whether grouping asthma subjects on the basis of BAL eosinophil % or neutrophil % could identify specific cytokine profiles. The only differences between neutrophil-normal asthma (neutrophil≤2.4%) and neutrophil-high asthma (neutrophils%>2.4%) were a higher BAL fluid IL-8 levels, and a lower FEV1 in the latter group. By contrast, compared to eosinophil-normal asthma (eosinophils≤0.3%), eosinophil-high asthma (eosinophils>0.3%) had higher levels of IL-5, IL-13, IL-16, and PDGF-bb, but same neutrophil percentage, IL-8, and FEV1. Our results identify neutrophils and IL-8 are the only inflammatory components in BAL fluids that distinguish controlled asthma from uncontrolled asthma, and both correlate inversely with FEV1.

Topics: Adult; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Case-Control Studies; Cytokines; Eosinophils; Female; Forced Expiratory Volume; Humans; Interleukin-5; Interleukin-8; Male; Neutrophils; Regression Analysis; Young Adult

Human airway smooth muscle cells (HASMC) contribute to asthma pathophysiology through an increased smooth muscle mass and elevated cytokine/chemokine output. Little is known about how HASMC and the airway epithelium interact to regulate chronic airway inflammation and remodeling. Amphiregulin is a member of the family of epidermal growth factor receptor (EGFR) agonists with cell growth and proinflammatory roles and increased expression in the lungs of asthma patients. Here we show that bradykinin (BK) stimulation of HASMC increases amphiregulin secretion in a mechanism dependent on BK-induced COX-2 expression, increased PGE2 output, and the stimulation of HASMC EP2 and EP4 receptors. Conditioned medium from BK treated HASMC induced CXCL8, VEGF, and COX-2 mRNA and protein accumulation in airway epithelial cells, which were blocked by anti-amphiregulin antibodies and amphiregulin siRNA, suggesting a paracrine effect of HASMC-derived amphiregulin on airway epithelial cells. Consistent with this, recombinant amphiregulin induced CXCL8, VEGF, and COX-2 in airway epithelial cells. Finally, we found that conditioned media from amphiregulin-stimulated airway epithelial cells induced amphiregulin expression in HASMC and that this was dependent on airway epithelial cell COX-2 activity. Our study provides evidence of a dynamic axis of interaction between HASMC and epithelial cells that amplifies CXCL8, VEGF, COX-2, and amphiregulin production.

Topics: Amphiregulin; Asthma; Bradykinin; Cells, Cultured; Cyclooxygenase 2; Dinoprostone; EGF Family of Proteins; Epithelial Cells; Gene Expression; Humans; Interleukin-8; Myocytes, Smooth Muscle; Respiratory Mucosa; Signal Transduction; Transcriptional Activation; Vascular Endothelial Growth Factor A

Bronchial epithelial cells represent the first line of defense against microorganisms and allergens in the airways and play an important role in chronic inflammatory processes such as asthma. In an experimental model, both RvD1 and AT-RvD1, lipid mediators of inflammation resolution, ameliorated some of the most important phenotypes of experimental asthma. Here, we extend these results and demonstrate the effect of AT-RvD1 on bronchial epithelial cells (BEAS-2B) stimulated with IL-4. AT-RvD1 (100 nM) decreased both CCL2 and CXCL-8 production, in part by decreasing STAT6 and NF-κB pathways. Furthermore, the effects of AT-RvD1 were ALX/FRP2 receptor dependent, as the antagonist of this receptor (BOC1) reversed the inhibition of these chemokines by AT-RvD1. In addition, AT-RvD1 decreased SOCS1 and increased SOCS3 expression, which play important roles in Th1 and Th17 modulation, respectively. In conclusion, AT-RvD1 demonstrated significant effects on the IL-4-induced activation of bronchial epithelial cells and consequently the potential to modulate neutrophilic and eosinophilic airway inflammation in asthma. Taken together, these findings identify AT-RvD1 as a potential proresolving therapeutic agent for allergic responses in the airways.

Topics: Asthma; Bronchi; Cell Line; Chemokine CCL2; Docosahexaenoic Acids; Epithelial Cells; Gene Expression; Humans; Interleukin-4; Interleukin-8; NF-kappa B; STAT6 Transcription Factor; Suppressor of Cytokine Signaling 1 Protein; Suppressor of Cytokine Signaling 3 Protein; Suppressor of Cytokine Signaling Proteins

Chronic inflammatory airway diseases, such as bronchial asthma and chronic obstructive pulmonary disease, are common respiratory disorders worldwide. Exacerbations of these diseases are frequent and worsen patients' respiratory condition and overall health. However, the mechanisms of exacerbation have not been fully elucidated. Recently, it was reported that interleukin (IL)-17A might play an important role in neutrophilic inflammation, which is characteristic of such exacerbations, through increased production of neutrophil chemoattractants. Therefore, we hypothesized that IL-17A was involved in the pathogenesis of acute exacerbation, due to viral infection in chronic inflammatory airway diseases. In this study, we assessed chemokine production by bronchial epithelial cells and investigated the underlying mechanisms. Comprehensive chemokine analysis showed that, compared with poly(I:C) alone, co-stimulation of BEAS-2B cells with IL-17A and poly(I:C) strongly induced production of such neutrophil chemoattractants as CXC chemokine ligand (CXCL)8, growth-related oncogene (GRO), and CXCL1. Co-stimulation synergistically induced CXCL8 and CXCL1 mRNA and protein production by BEAS-2B cells and normal human bronchial epithelial cells. Poly(I:C) induced chemokine expression by BEAS-2B cells mainly via Toll-like receptor 3/TIR-domain-containing adapter-inducing interferon-β-mediated signals. The co-stimulation with IL-17A and poly(I:C) markedly activated the p38 and extracellular-signal-regulated kinase 1/2 pathway, compared with poly(I:C), although there was little change in nuclear factor-κB translocation into the nucleus or the transcriptional activities of nuclear factor-κB and activator protein 1. IL-17A promoted stabilization of CXCL8 mRNA in BEAS-2B cells treated with poly(I:C). In conclusion, IL-17A appears to be involved in the pathogenesis of chronic inflammatory airway disease exacerbation, due to viral infection by promoting release of neutrophil chemoattractants from bronchial epithelial cells.

Topics: Asthma; Bronchi; Chemokine CXCL1; Chemotactic Factors; Epithelial Cells; Humans; Interleukin-17; Interleukin-8; Ligands; Neutrophils; p38 Mitogen-Activated Protein Kinases; Poly I-C; Toll-Like Receptor 3

Steroid resistance is a significant problem in management of chronic inflammatory diseases, including asthma. Accessible biomarkers are needed to identify steroid resistant patients to optimize their treatment. This study examined corticosteroid resistance in severe asthma. 24 asthmatics with forced expiratory volume in one second of less then 80% predicted were classified as steroid resistant or steroid sensitive based on changes in their lung function following a week of treatment with oral prednisone. Heparinised blood was collected from patients prior to oral prednisone administration. Phosphorylated mitogen activated kinases (MAPK) (extracellular regulated kinase (ERK), p38 and jun kinase (JNK)) were analyzed in whole blood samples using flow cytometry. Activation of phospho-p38 MAPK and phospho-mitogen- and stress-activated protein kinase 1 (MSK1) in asthmatics' peripheral blood mononuclear cells (PBMC) were confirmed by Western blot. Dexamethasone suppression of the LPS-induced IL-8 mRNA production by steroid resistant asthmatics PBMC in the presence of p38 and ERK inhibitors was evaluated by real time PCR. Flow cytometry analysis identified significantly stronger p38 phosphorylation in CD14+ monocytes from steroid resistant than steroid sensitive asthmatics (p = 0.014), whereas no difference was found in phosphorylation of ERK or JNK in CD14+ cells from these two groups of asthmatics. No difference in phosphorylated p38, ERK, JNK was detected in CD4+, CD8+ T cells, B cells and NK cells from steroid resistant vs. steroid sensitive asthmatics. P38 MAPK pathway activation was confirmed by Western blot, as significantly higher phospho-p38 and phospho-MSK1 levels were detected in the PBMC lysates from steroid resistant asthmatics. P38 inhibitor significantly enhanced DEX suppression of LPS-induced IL-8 mRNA by PBMC of steroid resistant asthmatics. This is the first report demonstrating selective p38 MAPK pathway activation in blood monocytes of steroid resistant asthmatics, suggesting that p38 and MSK1 phosphorylation can serve as blood biomarkers of steroid resistance.

Topics: Adult; Asthma; Drug Resistance; Female; Humans; Interleukin-8; Male; Monocytes; p38 Mitogen-Activated Protein Kinases; Prednisone; Ribosomal Protein S6 Kinases, 90-kDa

Respiratory infections are a major cause of asthma exacerbations where neutrophilic inflammation dominates and is associated with steroid refractory asthma. Structural airway cells in asthma differ from nonasthmatics; however it is unknown if neutrophils differ. We investigated neutrophil immune responses in patients who have good (AGood) and suboptimal (ASubopt) asthma symptom control.. Peripheral blood neutrophils from AGood (ACQ < 0.75, n = 11), ASubopt (ACQ > 0.75, n = 7), and healthy controls (HC) (n = 9) were stimulated with bacterial (LPS (1 μg/mL), fMLF (100 nM)), and viral (imiquimod (3 μg/mL), R848 (1.5 μg/mL), and poly I:C (10 μg/mL)) surrogates or live rhinovirus (RV) 16 (MOI1). Cell-free supernatant was collected after 1 h for neutrophil elastase (NE) and matrix metalloproteinase- (MMP-) 9 measurements or after 24 h for CXCL8 release. Results. Constitutive NE was enhanced in AGood neutrophils compared to HC. fMLF stimulated neutrophils from ASubopt but not AGood produced 50% of HC levels. fMLF induced MMP-9 was impaired in ASubopt and AGood compared to HC. fMLF stimulated CXCL8 but not MMP-9 was positively correlated with FEV1 and FEV1/FVC. ASubopt and AGood responded similarly to other stimuli.. Circulating neutrophils are different in asthma; however, this is likely to be related to airflow limitation rather than asthma control.

Topics: Aged; Asthma; Female; Forced Expiratory Volume; Humans; Immunity, Innate; Interleukin-8; Male; Matrix Metalloproteinase 9; Middle Aged; Neutrophils; Vital Capacity

We hypothesised that increased oxidative stress, as present in the airways of asthma and chronic obstructive pulmonary disease (COPD) patients, induces epithelial damage and reduces epithelial responsiveness to suppressive effects of corticosteroids on proinflammatory cytokine production and barrier function.. We induced oxidative stress by H2O2 and/or cigarette smoke extract (CSE) in human bronchial epithelial 16HBE cells and primary bronchial epithelial cells (PBEC) derived by brushings from asthma patients, COPD patients, and smoking and non-smoking control individuals. We investigated effects of budesonide on barrier function (electrical resistance) and TNF-α-induced proinflammatory cytokine production (IL-8/CXCL8, granulocyte macrophage-colony stimulating factor (GM-CSF)).. We observed that H2O2 and CSE reduce epithelial resistance. Budesonide significantly counteracted this effect, likely by protection against epidermal growth factor receptor-dependent cell-cell contact disruption. Furthermore, budesonide suppressed proinflammatory cytokine production. H2O2 pretreatment reduced this effect of budesonide on cytokine production in both 16HBE cells and PBECs. Importantly, PBECs from asthma and COPD patients were less sensitive to budesonide with respect to cytokine production and barrier function than PBECs from control subjects.. Together, our data indicate that budesonide suppresses epithelial proinflammatory responses and barrier dysfunction and that oxidative stress reduces these effects in airway epithelium from asthma and COPD patients. Therefore, restoration of corticosteroid responsiveness in asthma and COPD may act to improve the airway epithelial barrier.

Topics: Adult; Asthma; Bronchi; Budesonide; Cells, Cultured; Epithelial Cells; Female; Glucocorticoids; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Male; Middle Aged; Oxidative Stress; Phosphorylation; Pulmonary Disease, Chronic Obstructive; Smoking

Parental smoking is known to worsen asthma symptoms in children and to make them refractory to asthma treatment, but the molecular mechanism is unclear. Oxidative stress from tobacco smoke has been reported to impair histone deacetylase-2 (HDAC2) via phosphoinositide-3-kinase (PI3K)/Akt activation and, thus, to reduce corticosteroid sensitivity. The aim of this study was to investigate passive smoking-dependent molecular abnormalities in alveolar macrophages (AMs) by comparing passive smoke-exposed children and non-passive smoke-exposed children with uncontrolled severe asthma.. BAL fluid (BALF) was obtained from 19 children with uncontrolled severe asthma (10 non-passive smoking-exposed subjects and nine passive smoking-exposed subjects), and HDAC2 expression/activity, Akt/HDAC2 phosphorylation levels, and corticosteroid responsiveness in AMs were evaluated.. Parental smoking reduced HDAC2 protein expression by 54% and activity by 47%, with concomitant enhancement of phosphorylation of Akt1 and HDAC2. In addition, phosphorylation levels of Akt1 correlated positively with HDAC2 phosphorylation levels and negatively with HDAC2 activity. Furthermore, passive smoke exposure reduced the inhibitory effects of dexamethasone on tumor necrosis factor-α-induced CXCL8 release in AMs. There were relatively higher neutrophil counts and CXCL8 concentrations in BALF and lower Asthma Control Test scores compared with non-passive smoke-exposed children with uncontrolled severe asthma.. Passive smoking impairs HDAC2 function via PI3K signaling activation, which could contribute to corticosteroid-insensitive inflammation in children with severe asthma. This novel mechanism will be a treatment target in children with severe asthma and stresses the need for a smoke-free environment for asthmatic children.

Topics: Adolescent; Asthma; Case-Control Studies; Child; Female; Glucocorticoids; Histone Deacetylase 2; Humans; Interleukin-8; Male; Oxidative Stress; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Severity of Illness Index; Signal Transduction; Tobacco Smoke Pollution

Rhinoviruses (RV) are the most common acute triggers of asthma, and airway epithelial cells are the primary site of infection. Asthmatic bronchial epithelial cells (BECs) have been found to have impaired innate immune responses to RV. RV entry and replication is recognized by pathogen recognition receptors (PRRs), specifically toll-like receptor (TLR)3 and the RNA helicases; retinoic acid-inducible gene I (RIG-I) and melanoma differentiation-associated gene 5 (MDA5).. Our aim was to assess the relative importance of these PRRs in primary bronchial epithelial cells (pBEC) from healthy controls and asthmatics following RV infection and determine whether deficient innate immune responses in asthmatic pBECs were due to abnormal signalling via these PRRs.. The expression patterns and roles of TLR3 and MDA5 were investigated using siRNA knock-down, with subsequent RV1B infection in pBECs from each patient group. We also used BX795, a specific inhibitor of TBK1 and IKKi.. Asthmatic pBECs had significantly reduced release of IL-6, CXCL-8 and IFN-λ in response to RV1B infection compared with healthy pBECs. In healthy pBECs, siMDA5, siTLR3 and BX795 all reduced release of IL-6, CXCL-10 and IFN-λ to infection. In contrast, in asthmatic pBECs where responses were already reduced, there was no further reduction in IL-6 and IFN-λ, although there was in CXCL-10.. Impaired antiviral responses in asthmatic pBECs are not due to deficient expression of PRRs; MDA5 and TLR3, but an inability to later activate types I and III interferon immune responses to RV infection, potentially increasing susceptibility to the effects of RV infection.

Topics: Adult; Asthma; Case-Control Studies; DEAD Box Protein 58; DEAD-box RNA Helicases; Epithelial Cells; Female; Gene Expression; Gene Knockdown Techniques; Humans; Interferon Regulatory Factor-3; Interferon-Induced Helicase, IFIH1; Interferons; Interleukin-6; Interleukin-8; Male; Middle Aged; Picornaviridae Infections; Receptors, Immunologic; Respiratory Mucosa; Rhinovirus; Signal Transduction; Toll-Like Receptor 3

Intermediary quantitative traits are a possible alternative for the identification of disease genes. This may be particularly relevant when diagnostic criteria are not very well defined as described for asthma. We analyzed serum samples from 944 individuals of 218 asthma families for 17 cytokines (eotaxin, GM-CSF, IFNγ, IL1B, IL1RA, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12(p40), IL-13, IL-17, IL-23, IL-33, TSLP and TNF-α) and determined the heritability. Linked chromosomal regions were identified by a genome-wide analysis using 334 autosomal microsatellite marker and association tested by further 550 SNP marker at genes implicated earlier with immune response. Heritability varied with TNF-α and IL-8 levels having the highest and TSLP having the lowest heritability. Linkage was significantly increased only for IL-12(p40) at D17S949. There were multiple significant single-nucleotide polymorphisms (SNP) associations (P<0.05) as found in the transmission disequilibrium test, whereas only a few replicated in parents or children only. These include SNPs in IL1RN that were associated with IL-33 and TSLP levels, and a SNP in NR3C2 that was associated with eotaxin, IL-13 and IFN-γ levels. Circulating level of serum cytokines exhibits genetic associations with asthma traits that are otherwise not detected using clinical diagnosis or when the clinical details are ambiguous.

Topics: Adult; Asthma; Child; Cytokines; Female; Genetic Predisposition to Disease; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-8; Linkage Disequilibrium; Male; Microsatellite Repeats; Pedigree; Polymorphism, Single Nucleotide; Quantitative Trait, Heritable; Receptors, Mineralocorticoid; Thymic Stromal Lymphopoietin; Tumor Necrosis Factor-alpha

Mesenchymal cells (fibroblasts) of the airway wall respond to cholinergic stimulation by releasing pro-inflammatory and chemotactic cytokines and may thus contribute to chronic inflammation of the lung. Here, we studied the anti-inflammatory potential of olodaterol, a long acting β2-adrenergic receptor agonist, and tiotropium, a long-acting muscarinic receptor antagonist, and whether they interact at the level of the cyclic AMP dependent signaling pathway. Pulmonary fibroblasts of asthmatic (n = 9) and non-asthmatic (n = 8) subjects were stimulated with the muscarinic receptor agonist carbachol and interleukin-1β (IL-1 beta) in presence or absence of tiotropium or olodaterol alone, or their combination. We also measured cAMP levels and phosphorylation of the cAMP response element binding protein (CREB). As single components, carbachol, olodaterol and tiotropium did not affect IL-6 and IL-8 release. Carbachol concentration-dependently enhanced the production of IL-1β-induced IL-6 and IL-8, which was blocked by the simultaneous addition of tiotropium. The combination of olodaterol plus tiotropium further reduced IL-6 and IL-8 release. Olodaterol induced cAMP and the phosphorylation of CREB, an effect counteracted by carbachol, but rescued by tiotropium. We conclude that olodaterol plus tiotropium cooperate to decrease the inflammatory response in pulmonary fibroblasts in vitro.

Topics: Adrenergic beta-2 Receptor Agonists; Adult; Aged; Anti-Inflammatory Agents; Asthma; Benzoxazines; Bronchodilator Agents; Carbachol; Cyclic AMP; Dose-Response Relationship, Drug; Drug Therapy, Combination; Female; Fibroblasts; Humans; In Vitro Techniques; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Middle Aged; Muscarinic Antagonists; Phosphorylation; Scopolamine Derivatives; Signal Transduction; Tiotropium Bromide

Th17-mediated neutrophilic airway inflammation has been implicated in decreased response to glucocorticoids in asthma. We aimed to investigate the effect of glucocorticoids on the airway epithelial release of the neutrophilic and Th17-cell chemoattractant CCL20. We studied CCL20 and CXCL8 sputum levels in asthmatic subjects using inhaled glucocorticoids or not, and the effect of budesonide on CCL20 and CXCL8 production in primary bronchial epithelial cells. The mechanism behind the effect of budesonide-induced CCL20 production was studied in 16HBE14o- cells using inhibitors for the glucocorticoid receptor, intracellular pathways and metalloproteases. We observed higher levels of CCL20, but not CXCL8, in the sputum of asthmatics who used inhaled glucocorticoids. CCL20 levels correlated with inhaled glucocorticoid dose and sputum neutrophils. Budesonide increased tumour necrosis factor (TNF)-α-induced CCL20 by primary bronchial epithelium, while CXCL8 was suppressed. In 16HBE14o- cells, similar effects were observed at the CCL20 protein and mRNA levels, indicating transcriptional regulation. Although TNF-α-induced CCL20 release was dependent on the ERK, p38 and STAT3 pathways, the increase by budesonide was not. Inhibition of glucocorticoid receptor or ADAM17 abrogated the budesonide-induced increase in CCL20 levels. We show that glucocorticoids enhance CCL20 production by bronchial epithelium, which may constitute a novel mechanism in Th17-mediated glucocorticoid-insensitive inflammation in asthma.

Topics: ADAM Proteins; ADAM17 Protein; Adult; Aged; Asthma; Budesonide; Cells, Cultured; Chemokine CCL20; Epithelial Cells; Epithelium; Extracellular Signal-Regulated MAP Kinases; Female; Glucocorticoids; Humans; Inflammation; Interleukin-8; Male; Metalloproteases; Middle Aged; Neutrophils; Receptors, Glucocorticoid; Sputum; STAT3 Transcription Factor; Th17 Cells; Tumor Necrosis Factor-alpha

Asian dust storms (ADS) contain various airborne particles that may augment airway inflammation by increasing the level of interleukin-8. The objective of the study was to investigate the association of exposure to an ADS with worsening of symptoms of adult asthma and the effect of ADS particles on interleukin-8 transcriptional activity.. The subjects were 112 patients with mild to moderate asthma who recorded scores for their daily upper and lower respiratory tract symptoms and measured morning peak expiratory flow (PEF) from March to May 2011. Interleukin-8 transcriptional activity was assessed in THP-G8 cells that were exposed to airborne particles collected during days of ADS exposure.. Of the 112 patients, 31 had comorbid allergic rhinitis (AR) and/or chronic sinusitis (CS), and had worsened scores for upper respiratory tract symptoms on ADS days compared to non-ADS days. Scores for lower respiratory tract symptoms during ADS days were higher than non-ADS days in all patients. Three patients also had unscheduled hospital visits for exacerbation of asthma on ADS days. However, there was no significant difference in daily morning PEF between ADS and non-ADS days. Airborne particles collected on ADS days induced interleukin-8 transcriptional activity in THP-G8 cells compared to the original soil of the ADS.. Exposure to an ADS aggravates upper and lower tract respiratory symptoms in patients with adult asthma. ADS airborne particles may increase airway inflammation through enhancement of interleukin-8 transcriptional activity.

Topics: Aged; Air Pollutants; Asthma; Dust; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Japan; Luciferases; Male; Middle Aged; Particulate Matter; Peak Expiratory Flow Rate; Rhinitis, Allergic; Rhinitis, Allergic, Perennial; Sinusitis; Wind

Allergic asthma can cause airway structural remodeling, involving the accumulation of extracellular matrix and thickening of smooth muscle. Tumor necrosis factor (TNF) family ligand LIGHT (TNFSF14) is a cytokine that binds herpesvirus entry mediator (HVEM)/TNFRSF14 and lymphotoxin β receptor (LTβR). LIGHT induces asthmatic cytokine IL-13 and fibrogenic cytokine transforming growth factor-β release from allergic asthma-related eosinophils expressing HVEM and alveolar macrophages expressing LTβR, respectively, thereby playing crucial roles in asthmatic airway remodeling. In this study, we investigated the effects of LIGHT on the coculture of human basophils/eosinophils and bronchial epithelial BEAS-2B cells. The expression of adhesion molecules, cytokines/chemokines, and matrix metalloproteinases (MMP) was measured by flow cytometry, multiplex, assay or ELISA. Results showed that LIGHT could significantly promote intercellular adhesion, cell surface expression of intercellular adhesion molecule-1, release of airway remodeling-related IL-6, CXCL8, and MMP-9 from BEAS-2B cells upon interaction with basophils/eosinophils, probably via the intercellular interaction, cell surface receptors HVEM and LTβR on BEAS-2B cells, and extracellular signal-regulated kinase, p38 mitogen activated protein kinase, and NF-κB signaling pathways. The above results, therefore, enhance our understanding of the immunopathological roles of LIGHT in allergic asthma and shed light on the potential therapeutic targets for airway remodeling.

Topics: Asthma; Basophils; Bronchi; Cell Adhesion; Cell Line; Cell Membrane; Coculture Techniques; Eosinophils; Epithelial Cells; Humans; Hypersensitivity; Interleukin-6; Interleukin-8; Macrophages; Matrix Metalloproteinase 9; Microscopy, Fluorescence; Recombinant Proteins; Signal Transduction; Tumor Necrosis Factor Ligand Superfamily Member 14

This study aims to observe the influence of 1,25-(OH)2D3 on airway inflammation and chemokine expression in asthmatic rats and to explore its significance in the treatment of asthma.. Wistar rats were randomly divided into a normal control group (N), an asthma group (A), a 1,25-(OH)2D3 group (VD), a budesonide group (P) and a 1,25-(OH)2D3 + budesonide treatment group (L). The acute asthma models were established through ovalbumin sensitisation and challenge. Lung tissues were stained with haematoxylin and eosin to observe pathologic changes, whereas an enzyme-linked immunosorbent assay was used to examine serum IgE, as well as the eosinophil chemoattractant protein (eotaxin) and interleukin-8 (IL-8) expression levels in bronchoalveolar lavage fluid and the serum.. VD treatment partially reversed the characteristic pathological changes of airway inflammation. The IgE, eotaxin, and IL-8 expression levels in the VD group were significantly lower than those in the A group (p < 0.05) but remained higher than those in the control group (p < 0.05).. 1,25-(OH)2D3 reduces airway inflammation, airway hyperresponsiveness and airway remodeling by partially inhibiting chemokine production during airway inflammation, and 1,25-(OH)2D3 synergises with hormone therapy.

Topics: Allergens; Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Calcitriol; Cell Count; Chemokine CCL11; Female; Immunoglobulin E; Interleukin-8; Lung; Ovalbumin; Rats, Wistar; Vitamins

Topics: Adult; Asthma; Bronchi; Cells, Cultured; Epithelial Cells; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; In Vitro Techniques; Interleukin-8; Stress, Mechanical; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

The aim of this study was to estimate relations between sputum neutrophilia and the chemotactic activity of peripheral blood neutrophils after the bronchial allergen challenge in asthma patients.. Fifteen patients with allergic asthma (AA), 13 patients with allergic rhinitis (AR), all sensitized to Dermatophagoides pteronyssinus, and 8 healthy subjects (HS) underwent bronchial challenge with D. pteronyssinus. Sputum and peripheral blood collection were performed 24 h before, 7 and 24 h after the bronchial challenge. Cell counts were determined by the May-Grünwald-Giemsa method. Neutrophil chemotaxis was analyzed by a flow cytometer; IL-8 levels were measured by ELISA.. Sputum neutrophil count and peripheral blood neutrophil chemotaxis of patients with AA were greater 7 and 24 h after the challenge compared with the baseline values and patients with AR and HS (P < 0.05). Moreover, a significant correlation was found between the neutrophil count in sputum and IL-8 levels, and the chemotactic activity of peripheral blood neutrophils 24 h after the bronchial challenge only the patients with AA (P < 0.05).. Increased sputum neutrophil count was found to be associated with an enhanced chemotactic activity of peripheral blood neutrophils during allergen-induced late-phase airway inflammation in patients with allergic asthma.

Topics: Adult; Allergens; Animals; Asthma; Bronchial Provocation Tests; Chemotaxis, Leukocyte; Dermatophagoides pteronyssinus; Female; Humans; Interleukin-8; Leukocyte Count; Male; Middle Aged; Neutrophil Infiltration; Neutrophils; Rhinitis, Allergic; Sputum; Young Adult

Infiltration of fibrocytes (FC) in the airway smooth muscle is a feature of asthma, but the pathological significance is unknown.. We sought to explore whether FC modulate the phenotype of airway smooth muscle cells (ASMC) in asthmatic vs. control subjects.. Fibrocytes were isolated from CD14+ monocytes from asthmatic and normal subjects. Proliferation of ASMC of asthmatic or normal subjects was analysed by (3) H-thymidine incorporation, cell number counting and Ki-67 expression after treatment of ASMC with FC-conditioned medium (FCCM) or co-culture with FC. ASMC-associated cytokines/chemokines implicated in asthma (TGF-β1, eotaxin, IL-6 and IL-8) were measured in co-culture or transwell culture of ASMC + FC by ELISA. Immunofluorescence staining was performed to localize these cytokines in ASMC. Cytokine secretion was measured in the transwell culture of ASMC + FC, where NF-κB-p65 or ERK1/2 in ASMC was silenced by siRNA. Contractile phenotype of ASMC in transwell culture was assessed by immunoblotting of α-smooth muscle actin (α-SMA) and myosin light chain kinase (MLCK).. Fibrocytes did not affect ASMC proliferation and expression of TGF-β1, eotaxin, α-SMA and MLCK; however, ASMC production of IL-8 and IL-6 was increased in the co-culture and transwell culture by FC. ASMC treated with FCCM were immunopositive for IL-8/IL-6 and produced more IL-8/IL-6. Furthermore, siRNA silencing of NF-κB-p65 or ERK1/2 in transwell cultures of asthmatic ASMC with normal subject FC decreased IL-8 and IL-6 production.. Fibrocytes promoted IL-8 and IL-6 production by ASMC, demonstrating a proinflammatory role for FC and a possible mechanism of the inflammatory phenotype in asthma.

Topics: Actins; Adult; Asthma; Case-Control Studies; Cell Communication; Cell Differentiation; Cells, Cultured; Coculture Techniques; Collagen Type I; Female; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Monocytes; Myocytes, Smooth Muscle; Myosin-Light-Chain Kinase; Phenotype; Signal Transduction

IL-17 is a pro-inflammatory mediator that is believed to play a critical role in regulating tissue inflammation during asthma, COPD, as well as other inflammatory disorders. The level of expression of IL-17 has been shown to be upregulated in lung bronchial tissue of asthmatic patients. Several reports have provided further evidence that this cytokine could play a key role in enhancing the migration of inflammatory as well as structural cells of the bronchial lung tissue during asthma and COPD. B cell infiltration to sites of inflammation during inflammatory disorders such as bowel disease, asthma and COPD has been reported. Accordingly, in this study we hypothesized that IL-17 may exert a chemotactic effect on primary B cells during asthma. We observed that B cells from asthmatic patients expressed significantly higher levels of IL-17RA and IL-17RC, compared to those of healthy subjects. Using an in-vitro migration assay, B cells were shown to migrate towards both IL-17A and IL-17F. Interestingly, blocking IL-17A and IL-17F signaling using either anti-IL-17R antibodies or MAP kinase inhibitors prevented in vitro migration of B cell towards IL-17. These observations indicate a direct chemotactic effect of IL-17 cytokines on primary peripheral blood B cells with higher effect being on asthmatic B cells. These findings revealed a key role for IL-17 in enhancing the migration of B cells to the lung tissue during asthma or COPD.

Topics: Adult; Asthma; B-Lymphocytes; Bronchi; Chemokine CXCL13; Chemotaxis; Enzyme Activation; Female; Humans; Imidazoles; Inflammation; Interleukin-17; Interleukin-8; Male; p38 Mitogen-Activated Protein Kinases; Pulmonary Disease, Chronic Obstructive; Pyridines; Receptors, Interleukin; Receptors, Interleukin-17; Th17 Cells

Asthma-related mortality has been decreasing due to inhaled corticosteroid use, but severe asthma remains a major clinical problem. One characteristic of severe asthma is resistance to steroid therapy, which is related to neutrophilic inflammation. Recently, the tumor necrosis factor superfamily member (TNFSF) 14/LIGHT has been recognized as a key mediator in severe asthmatic airway inflammation. However, the profiles and intracellular mechanisms of cytokine/chemokine production induced in cells by LIGHT are poorly understood. We aimed to elucidate the molecular mechanism of LIGHT-induced cytokine/chemokine production by bronchial epithelial cells. Human bronchial epithelial cells express lymphotoxin β receptor (LTβR), but not herpesvirus entry mediator, which are receptors for LIGHT. LIGHT induced various cytokines/chemokines, such as interleukin (IL)-6, oncostatin M, monocyte chemotactic protein-1, growth-regulated protein α and IL-8. Specific siRNA for LTβR attenuated IL-6 and IL-8 production by BEAS-2B and normal human bronchial epithelial cells. LIGHT activated intracellular signaling, such as mitogen-activated protein kinase and nuclear factor-κB (NF-κB) signaling. LIGHT also induced luciferase activity of NF-κB response element, but not of activator protein-1 or serum response element. Specific inhibitors of phosphorylation of extracellular signal-regulated kinase (Erk) and that of inhibitor κB attenuated IL-8 production, suggesting that LIGHT-LTβR signaling induces IL-8 production via the Erk and NF-κB pathways. LIGHT, via LTβR signaling, may contribute to exacerbation of airway neutrophilic inflammation through cytokine and chemokine production by bronchial epithelial cells.

Topics: Asthma; Bronchi; Epithelial Cells; Humans; Inflammation; Interleukin-6; Interleukin-8; Lymphotoxin beta Receptor; MAP Kinase Signaling System; Mitogen-Activated Protein Kinases; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Signal Transduction; Tumor Necrosis Factor Ligand Superfamily Member 14; Tumor Necrosis Factor-alpha

To explore the effects of smoking on airway inflammation through the expressions of hypoxia-inducible factor-1α (HIF-1α) and histone deacetylase-2 (HDAC2) in asthmatic mice.. A total of 30 female SPF BALB/c mice were divided randomly into 3 groups of control (C), asthma (A) and smoking asthma (S). The latter two groups were sensitized and challenged with ovalbumin (OVA) for asthmatic modeling. The mice of group S were placed into a self-made fumigating box for passive smoking. While group S was sensitized and challenged with normal saline instead of OVA. The pathological changes of different groups were observed. The different cell counts of bronchoalveolar fluid (BALF) were analyzed. The expressions of HIF-1α and HDAC2 were detected by immunohistochemistry. The level of interleukin (IL)-8 in BALF was detected by enzyme-linked immunosorbent assay (ELISA). And the levels of HIF-1α and HDAC2 in lung homogenate were measured by Western blot.. The ratios of eosinophil (EOS) to total cell numbers of BALF in groups A and S were significantly higher than that in group C ((8.90 ± 1.60)%, (7.52 ± 0.63)% vs (0.60 ± 0.10)%, both P < 0.01), while the ratio of neutrophile (NEU) in group S was higher than that in group A ((18.24 ± 5.19)% vs (8.46 ± 1.58)%, P < 0.01). Western blot showed that HIF-1α expressions in lung homogenate of groups A and S were significantly elevated than that in groups C (0.144 ± 0.008, 0.238 ± 0.015 vs 0.081 ± 0.005, both P < 0.01). While the HIF-1α level of group S was higher than that of group A (P < 0.01). And the expressions of HDAC2 in groups A and S significantly decreased than that in group C (0.287 ± 0.008, 0.164 ± 0.015 vs 0.452 ± 0.041, both P < 0.01). While the HDAC2 level of group S was lower than that of group A (P < 0.01). The BALF level of IL-8 in group S was higher than those in groups A and C ((42.07 ± 4.54) vs (21.66 ± 2.78), (14.33 ± 3.73) pg/ml, both P < 0.01). There were significantly negative correlations between the expressions of HIF-1α and HDAC2 (r = -0.950, P < 0.01) in lung as well as HDAC2 in lung and IL-8 (r = -0.855, P < 0.01) in BALF.. Cigarette smoking aggravates the airway inflammation through a down-regulated expression of HDAC2 by activating HIF-1α.

Topics: Animals; Asthma; Enzyme-Linked Immunosorbent Assay; Eosinophils; Female; Histone Deacetylase 2; Hypoxia-Inducible Factor 1, alpha Subunit; Interleukin-8; Lung; Mice; Mice, Inbred BALB C; Smoking

The collection of exhaled breath condensate (EBC) is a noninvasive method that can be used to monitor the inflammatory status of patients with chronic airway diseases. We aimed to study differences in cytokine expression between patients with exacerbations of chronic obstructive pulmonary disease (COPD) and patients with asthma attacks.. Using a custom-made device and methods based on American Thoracic Society (ATS)/European Respiratory Society (ERS) recommendations, EBC samples were collected from nine COPD patients, 12 asthma patients and 10 healthy individuals. Cytokine concentrations in serum and EBC were measured via commercial ELISA kits.. Of four cytokines measured in EBC [interleukin-8 (IL-8), IL-17, IL-4 and tumor necrosis factor-α (TNF-α)], only IL-8 was significantly higher in COPD than in asthma patients (5.27 ± 0.18 vs. 4.36 ± 0.34 pg/mL, p = 0.001). Moreover, COPD patients had higher serum IL-8 than asthma patients (10.57 ± 0.55 vs. 5.15 ± 0.24 pg/mL, p < 0.001). No significant correlation between serum and EBC cytokine concentrations was observed in each subgroup of patients.. Compared with patients with asthma attacks, patients with exacerbated COPD had increased IL-8 expression in both serum and EBC. These results suggest that IL-8 may be more important in airway and systemic inflammation in COPD exacerbations than in asthma attacks.

Topics: Adult; Aged; Asthma; Breath Tests; Disease Progression; Female; Humans; Interleukin-8; Male; Middle Aged; Pulmonary Disease, Chronic Obstructive

Although it is recognized that IL-33 plays a key role in the onset of asthma, it is currently unclear whether IL-33 acts on any other target cells besides mast cells and Th2 cells in asthma. We investigated that whether airway smooth muscle cells (ASMCs) could contribute to asthma via stimulation with IL-33.. To create a mouse model of acute asthma, murine ASMCs were isolated and cultured in vitro with IL-33. The ASMCs were divided into two groups, ASMCs from normal mice and ASMCs from ovalbumin-sensitized mice. The release of mouse KC was analyzed by PCR and ELISA. Immunocytochemical Staining of murine ASMCs for ST2 and IL-1RAcP was performed.. IL-33 promoted KC expression, both in terms of mRNA and protien levels, in ASMCs from ovalbumin-sensitized mice. ST2 and IL-1RAcP were expressed in the membrane of ASMCs in ovalbumin-sensitized mice.. IL-33 may contribute to the inflammation in the airways by acting on airway smooth muscle cells. IL-33 and ST2 may play important roles in allergic bronchial asthma.

Topics: Animals; Asthma; Cells, Cultured; Chemokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Immunohistochemistry; Inflammation; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Interleukin-8; Interleukins; Lung; Mice; Mice, Inbred BALB C; Myocytes, Smooth Muscle; Ovalbumin; Real-Time Polymerase Chain Reaction; Receptors, Interleukin

Thymic stromal lymphoproetin (TSLP) is a cytokine secreted by the airway epithelium in response to respiratory viruses and it is known to promote allergic Th2 responses in asthma. This study investigated whether virally-induced secretion of TSLP is directional in nature (apical vs. basolateral) and/or if there are TSLP-mediated effects occurring at both sides of the bronchial epithelial barrier in the asthmatic state.. Primary human bronchial epithelial cells (HBEC) from control (n = 3) and asthmatic (n = 3) donors were differentiated into polarized respiratory tract epithelium under air-liquid interface (ALI) conditions and treated apically with dsRNA (viral surrogate) or TSLP. Sub-epithelial effects of TSLP were examined in human airway smooth muscle cells (HASMC) from normal (n = 3) and asthmatic (n = 3) donors. Clinical experiments examined nasal airway secretions obtained from asthmatic children during naturally occurring rhinovirus-induced exacerbations (n = 20) vs. non-asthmatic uninfected controls (n = 20). Protein levels of TSLP, CCL11/eotaxin-1, CCL17/TARC, CCL22/MDC, TNF-α and CXCL8 were determined with a multiplex magnetic bead assay.. Our data demonstrate that: 1) Asthmatic HBEC exhibit an exaggerated apical, but not basal, secretion of TSLP after dsRNA exposure; 2) TSLP exposure induces unidirectional (apical) secretion of CCL11/eotaxin-1 in asthmatic HBEC and enhanced CCL11/eotaxin-1 secretion in asthmatic HASMC; 3) Rhinovirus-induced asthma exacerbations in children are associated with in vivo airway secretion of TSLP and CCL11/eotaxin-1.. There are virally-induced TSLP-driven secretory immune responses at both sides of the bronchial epithelial barrier characterized by enhanced CCL11/eotaxin-1 secretion in asthmatic airways. These results suggest a new model of TSLP-mediated eosinophilic responses in the asthmatic airway during viral-induced exacerbations.

Topics: Adolescent; Asthma; Case-Control Studies; Cell Line; Chemokine CCL11; Chemokine CCL17; Chemokine CCL22; Child; Child, Preschool; Cytokines; Humans; Interleukin-8; Nasal Mucosa; Rhinovirus; RNA, Double-Stranded; Thymic Stromal Lymphopoietin; Tumor Necrosis Factor-alpha

Neutrophilic airway inflammation associated with multiple allergens has been related to steroid-resistant asthma. However, most animal models use only one allergen, which cannot simulate asthma closely as seen in patients. To determine the mechanism of inflammatory process involved in this severe condition, BALB/c mice were repetitively challenged with the pooled extract of dust mite, ragweed, and Aspergillus species (DRA). We found that DRA increased interleukin (IL)-10 and TGF-β levels and neutrophil recruitment in bronchial alveolar lavage fluid. We also found that although dexamethasone suppressed the release of these two cytokines, mast cells recruitment, and mucus hypersecretion, it actually increased neutrophil infiltration and the level of keratinocyte-derived chemokine (mKC), a functional homolog of human IL-8. Treatment of human lung alveolar A549 cells with Der p1, an extract of house dust mite Dermatophagoides pteronyssinus, increased the expression of IL-8 and activity of NF-κB. The elevated IL-8 level was suppressed by BAY11-7082, a selective NF-κB inhibitor, but not by dexamethasone. These results suggest that increased IL-8 (mKC) levels may be involved in steroid-resistant neutrophilic airway inflammation through an NF-κB-dependent pathway.

Topics: Animals; Antigens, Plant; Aspergillus; Asthma; Bronchoalveolar Lavage Fluid; Cell Line, Tumor; Dermatophagoides pteronyssinus; Dexamethasone; Female; Humans; Inflammation; Interleukin-10; Interleukin-8; Mice; Mice, Inbred BALB C; Neutrophil Infiltration; NF-kappa B; Plant Extracts; Transforming Growth Factor beta

Th17 cells and IL-17A play a role in the development and progression of allergic diseases. We analyzed the IL-17A levels in sputum supernatants (Ss), nasal wash (NW) and plasma (P) from Healthy Controls (HC) and children with Asthma/Rhinitis. We tested the expression of IL-17A, RORγ(t) and FOXP3 in peripheral blood T-lymphocytes from intermittent and mild-moderate asthma. The effect of Budesonide and Formoterol was tested "in vitro" on IL-17A, RORγ(t) and FOXP3 expression in cultured T-lymphocytes from mild-moderate asthma/persistent rhinitis patients, and on nasal and bronchial epithelial cells stimulated with NW and Ss from mild-moderate asthma/persistent rhinitis. Further, the effect of 12 weeks of treatment with Budesonide and Formoterol was tested "in vivo" in T-lymphocytes from mild-moderate asthma/persistent rhinitis patients. IL-17A was increased in Ss, NW and P from children with mild-moderate asthma compared with intermittent and HC. In cultured T-lymphocytes IL-17A and RORγ(t) expression were higher in mild-moderate asthma/persistent rhinitis than in mild-moderate asthma/intermittent rhinitis, while FOXP3 was reduced. Budesonide with Formoterol reduced IL-17A and RORγ(t), while increased FOXP3 in cultured T-lymphocytes from mild-moderate asthma/persistent rhinitis, and reduced the IL-8 release mediated by IL-17A present in NW and Ss from mild-moderate asthma/persistent rhinitis in nasal and bronchial epithelial cells. Finally, Budesonide with Formoterol reduced IL-17A levels in P and Ss, CD4(+)IL-17A(+)T-cells, in naïve children with mild-moderate asthma/persistent rhinitis after 12 weeks of treatment. Th17 mediated immunity may be involved in the airway disease of children with allergic asthma and allergic rhinitis. Budesonide with Formoterol might be a useful tool for its therapeutic control.

Topics: Administration, Inhalation; Adolescent; Adrenergic beta-2 Receptor Agonists; Anti-Asthmatic Agents; Asthma; Budesonide; Case-Control Studies; Cells, Cultured; Child; Ethanolamines; Female; Forkhead Transcription Factors; Formoterol Fumarate; Humans; Interleukin-17; Interleukin-8; Male; Nasal Lavage Fluid; Nuclear Receptor Subfamily 1, Group F, Member 3; Rhinitis, Allergic, Perennial; Sputum; Th17 Cells

Although inhaled glucocorticoids are the mainstays of asthma treatment, they are poorly effective at treating and preventing virus-induced asthma exacerbations. The major viruses precipitating asthma exacerbations are rhinoviruses.. We sought to evaluate whether rhinovirus infection interferes with the mechanisms of action of glucocorticoids.. Cultured primary human bronchial or transformed (A549) respiratory epithelial cells were infected with rhinovirus 16 (RV-16) before dexamethasone exposure. Glucocorticoid receptor (GR) α nuclear translocation, glucocorticoid response element (GRE) binding, and transactivation/transrepression functional readouts were evaluated by using immunocytochemistry, Western blotting, DNA binding assays, real-time quantitative PCR, coimmunoprecipitation, and ELISA techniques. Specific inhibitors of c-Jun N-terminal kinase (JNK) and of IκB kinase (IKK) were used to investigate the involvement of intracellular signaling pathways.. RV-16 infection impaired dexamethasone-dependent (1) inhibition of IL-1β-induced CXCL8 release, (2) induction of mitogen-activated protein kinase phosphatase 1 gene expression, and (3) binding of GR to GREs in airway epithelial cells. This was associated with impaired GRα nuclear translocation, as assessed by means of both immunochemistry (54.0% ± 6.8% vs 24.7% ± 3.8% GR-positive nuclei after 10 nmol/L dexamethasone treatment in sham- or RV-16-infected cells, respectively; P < .01) and Western blotting. RV-16 infection induced nuclear factor κB activation and GRα phosphorylation, which were prevented by inhibitors of IKK2 and JNK, respectively. In rhinovirus-infected cells the combination of JNK and IKK2 inhibitors totally restored dexamethasone suppression of CXCL8 release, induction of mitogen-activated protein kinase phosphatase 1 gene expression, and GRα nuclear translocation.. RV-16 infection of human airway epithelium induces glucocorticoid resistance. Inhibition of RV-16-induced JNK and nuclear factor κB activation fully reversed rhinovirus impairment of both GRα nuclear translocation and the transactivation/transrepression activities of glucocorticoids.

Topics: Asthma; Cell Line; Cell Nucleus; Dexamethasone; Drug Resistance; Enzyme Activation; Glucocorticoids; Humans; I-kappa B Kinase; Interleukin-1beta; Interleukin-8; JNK Mitogen-Activated Protein Kinases; NF-kappa B; Picornaviridae Infections; Protein Transport; Receptors, Glucocorticoid; Respiratory Mucosa; Rhinovirus

There have been concerns about the cardiovascular safety of omalizumab.. In this study, the clinical status of the omalizumab receiving severe asthma patients and the cytokine expressions patterns were investigated.. In a pilot study described below we examined the levels of serum eosinophil cationic peptid (ECP), CD200, d-dimer, 25-hydroxyvitamin D (25(OH)D), CXCL8 and IL-1β in asthma patients treated with anti-IgE therapy, to explore their relationship with disease activity, and the impact of anti-IgE therapy impact on those levels. Exercise stress testing and blood samples were taken at all follow up visits from the time of first diagnosis and after 20 months of treatment during the disease remission.. Fractional exhaled nitric oxide concentrations and serum levels of sTRAIL, sCD200, D-dimer, ECP, total IgE, IL-1β and Hs-CRP were decreased while CXCL8, 25(OH)D were increased after starting the treatment of anti-IgE. Our first case of a patient, who had both protein C and S deficiency and hence a high risk for thromboembolism, documents for the first time the safety of omalizumab for asthmatic patients with concurrent risk factors contributing to arteriothrombotic events.. Omalizumab might be used carefully in patients with cardiovascular diseases.

Topics: Antibodies, Anti-Idiotypic; Antibodies, Monoclonal, Humanized; Antigens, CD; Asthma; Cardiovascular Diseases; Female; Fibrin Fibrinogen Degradation Products; Humans; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Omalizumab; Vitamin D

Chronic exposure to ambient particulate matter pollution during childhood is associated with decreased lung function growth and increased prevalence of reported respiratory symptoms. The role of airway epithelium-derived factors has not been well determined.. To determine if urban particulate matter (UPM) stimulates production of vascular endothelial growth factor (VEGF) and transforming growth factor-β2 (TGF-β2), and gene expression of mucin 5AC (MUC5AC) and interleukin-(IL)-8 by primary airway epithelial cells (AECs) obtained from carefully phenotyped healthy and atopic asthmatic school-aged children.. Primary AECs from 9 healthy and 14 asthmatic children were differentiated in air--liquid interface (ALI) culture. The apical surface was exposed to UPM suspension or phosphate buffered saline (PBS) vehicle control for 96 h. VEGF and TGF-β2 concentrations in cell media at baseline, 48 and 96 h were measured via ELISA. MUC5AC and IL-8 expression by AECs at 96 h was measured via quantitative polymerase chain reaction.. Baseline concentrations of VEGF, but not TGF-β2, were significantly higher in asthmatic versus healthy cultures. UPM stimulated production of VEGF, but not TGF-β2, at 48 and 96 h; the magnitude of change was comparable across groups. At 96 h there was greater MUC5AC and IL-8 expression by UPM exposed compared to PBS exposed AECs.. Induction of the pro-remodeling cytokine VEGF may be a potential mechanism by which UPM influences lung function growth in children irrespective of asthma status. Respiratory morbidity associated with UPM exposure in children may be related to increased expression of MUC5AC and IL-8.

Topics: Adolescent; Air Pollutants; Airway Remodeling; Asthma; Cells, Cultured; Child; Cities; Epithelial Cells; Female; Gene Expression; Humans; Interleukin-8; Male; Mucin 5AC; Particulate Matter; Transforming Growth Factor beta2; Vascular Endothelial Growth Factor A

Airway wall remodelling is a key pathology of asthma. It includes thickening of the airway wall, hypertrophy and hyperplasia of bronchial smooth muscle cells (BSMC), as well as an increased vascularity of the sub-epithelial cell layer. BSMC are known to be the effector cells of bronchoconstriction, but they are increasingly recognized as an important source of inflammatory mediators and angiogenic factors.. To compare the angiogenic potential of BSMC of asthmatic and non-asthmatic patients and to identify asthma-specific angiogenic factors.. Primary BSMC were isolated from human airway tissue of asthmatic and non-asthmatic patients. Conditioned medium (CM) collected from BSMC isolates was tested for angiogenic capacity using the endothelial cell (EC)-spheroid in vitro angiogenesis assay. Angiogenic factors in CM were quantified using a human angiogenesis antibody array and enzyme linked immunosorbent assay.. Induction of sprout outgrowth from EC-spheroids by CM of BSMC obtained from asthma patients was increased compared with CM of control BSMC (twofold, p < 0.001). Levels of ENA-78, GRO-α and IL-8 were significantly elevated in CM of BSMC from asthma patients (p < 0.05 vs. non-asthmatic patients). SB 265610, a competitive antagonist of chemokine (CXC-motif) receptor 2 (CXCR2), attenuated the increased sprout outgrowth induced by CM of asthma patient-derived BSMC.. BSMC isolated from asthma patients exhibit increased angiogenic potential. This effect is mediated through the CXCR2 ligands (ENA78, GRO-α and IL-8) produced by BSMC.. CXCR2 ligands may play a decisive role in directing the neovascularization in the sub-epithelial cell layers of the lungs of asthma patients. Counteracting the CXCR2-mediated neovascularization by pharmaceutical compounds may represent a novel strategy to reduce airway remodelling in asthma.

Topics: Adult; Asthma; Bronchi; Chemokine CXCL1; Chemokine CXCL5; Chemokines, CXC; Female; Humans; Interleukin-8; Ligands; Male; Middle Aged; Myocytes, Smooth Muscle; Neovascularization, Pathologic; Phenylurea Compounds; Receptors, Interleukin-8B; Triazoles; Young Adult

A notable feature of allergic asthma is the infiltration of mast cells into smooth muscle in the human airway. Thus, mast cells and human airway smooth muscle (hASM) cells are likely to exhibit mutual functional modulation via direct cell-cell contact or through released factors. This study examined mast cell modulation of hASM cell cytokine release.. The mast cell line HMCα was used to model mast cell function. hASM cells were either co-cultured directly with resting or IgE/antigen-stimulated HMCα cells or treated with HMCα-conditioned media to examine the impact on cytokine release. The activation pathways triggered in hASM cells by the mast cell-derived factors were examined through the use of selective inhibitors and by Western blotting.. HMCα cells, or their conditioned media, induced the expression of cytokines (IL-8 and IL-6) by hASM cells at both the mRNA and the protein level. Cytokine expression in hASM cells was greatly amplified when HMCα cells were IgE/antigen-activated. The effects of the conditioned media were not mediated by the chemokines MCP-1 and MIP-1α or by exosomes. While the mast cell-derived factor(s) increased p38(MAPK) phosphorylation in hASM cells, cytokine production was not inhibited by the p38(MAPK) inhibitor SB203580. hASM cell production of IL-8 induced by HMCα condition media but not IL-6 was, however, attenuated by the Src tyrosine kinase inhibitor PP2.. Our study shows that the release of soluble mediators by activated mast cells can stimulate hASM cells to elicit production of proinflammatory cytokines that may then exacerbate airway inflammation in asthma.

Topics: Asthma; Cell Line; Culture Media, Conditioned; Exosomes; Humans; Imidazoles; Immunoglobulin E; Inflammation; Interleukin-6; Interleukin-8; Mast Cells; Myocytes, Smooth Muscle; p38 Mitogen-Activated Protein Kinases; Pyridines; Pyrimidines; Receptors, Fc; Respiratory System; RNA, Messenger; src-Family Kinases

Smoking may modify the inflammatory pattern of the asthmatic airways. Osteopontin (OPN) has been associated with inflammation and fibrosis. In asthma, sputum levels of OPN are elevated and have been related to the underlying severity and to mediators expressing remodeling and inflammation. To evaluate the levels of OPN in sputum supernatants of asthmatic patients and to investigate the possible role of smoking as well as associations with mediators and cells involved in the inflammatory and remodeling process. We studied 103 asthma patients (49 smokers) and 40 healthy subjects (20 smokers) who underwent lung function tests, bronchial hyperresponsiveness to methacholine, and sputum induction for cell count identification and measurement of OPN, TGF-β1, IL-8, IL-13 and ECP in sputum supernatants. The concentrations of all mediators were measured using enzyme immunoassays. OPN levels (pg/ml) were significantly higher in smoking asthmatics compared to non-smoking asthmatics, and both non-smoking and smoking controls [median (interquartile ranges) 1120 (651,1817) vs. 197 (118,341) vs. 50 (42,70) vs. 102 (77,110) pg/ml, respectively; p<0.001]. Regression analysis provided significant associations between OPN and sputum neutrophils, IL-8 and TGF-β1, the most significant being the one with TGF-β1. These associations were present only in smoking asthmatics. Smoking habit significantly affects sputum OPN levels in asthma. The associations of OPN with sputum neutrophils, TGF-β1 and IL-8 in smoking asthmatics suggest a possible role for OPN in the neutrophilic inflammation and remodeling process in this phenotype of asthma.

Topics: Asthma; Bronchial Hyperreactivity; Female; Humans; Inflammation Mediators; Interleukin-13; Interleukin-8; Male; Methacholine Chloride; Middle Aged; Neutrophils; Osteopontin; Respiratory Function Tests; Smoking; Sputum; Transforming Growth Factor beta1

The airway epithelium forms a physical, chemical and immunological barrier against inhaled environmental substances. In asthma, these barrier properties are thought to be abnormal. In this study, we analysed the effect of grass pollen on the physical and immunological barrier properties of differentiated human primary bronchial epithelial cells. Following exposure to Timothy grass (Phleum pratense) pollen extract, the integrity of the physical barrier was not impaired as monitored by measuring the transepithelial resistance and immunofluorescence staining of tight junction proteins. In contrast, pollen exposure affected the immunological barrier properties by modulating vectorial mediator release. CXC chemokine ligand (CXCL)8/interleukin (IL)-8 showed the greatest increase in response to pollen exposure with preferential release to the apical compartment. Inhibition of the extracellular signal-regulated kinase 1/2 and p38 mitogen-activated protein kinase pathways selectively blocked apical CXCL8/IL-8 release via a post-transcriptional mechanism. Apical release of CC chemokine ligand (CCL)20/macrophage inflammatory protein-3α, CCL22/monocyte-derived chemokine and tumour necrosis factor-α was significantly increased only in severe asthma cultures, while CCL11/eotaxin-1 and CXCL10/interferon-γ-induced protein-10 were reduced in nonasthmatic cultures. The bronchial epithelial barrier modulates polarised release of mediators in response to pollen without direct effects on its physical barrier properties. The differential response of cells from normal and asthmatic donors suggests the potential for the bronchial epithelium to promote immune dysfunction in asthma.

Topics: Allergens; Asthma; Bronchi; Bronchoscopy; Cells, Cultured; Chemokines; Epithelial Cells; Humans; Inflammation; Interleukin-8; Ligands; Plant Extracts; Poaceae; Pollen

Recently, the serum levels of YKL-40, a chitinase-like glycoprotein, have been shown to be significantly elevated in asthmatics and are associated with asthma severity. Although these studies raise the possibility that YKL-40 may influence asthma, the mechanisms remain unknown. This study firstly investigated the mechanisms involved in YKL-40-mediated inflammation in human bronchial epithelial cells (HBECs) and analyzed the soluble factors secreted by bronchial epithelial cells exposed to YKL-40 that were responsible for increasing proliferation and migration of primary normal human bronchial smooth muscle cells (BSMCs). YKL-40-induced inflammation was assayed in two HBECs (BEAS-2B cell line and primary HBECs). In addition, we treated BEAS-2B cells and HBECs with YKL-40 and added the conditioned culture media to BSMCs. The proliferation and migration of BSMCs were determined by premixed WST-1 cell proliferation reagent (Clontech Laboratories) and QCM chemotaxis migration assay (Millipore), respectively. Bronchial epithelial cells treated with YKL-40 resulted in a significant increase of IL-8 production, which was dependent on MAPK (JNK and ERK) and NF-κB pathways activation. YKL-40-induced IL-8 was found to further stimulate proliferation and migration of BSMCs, and the effects were inhibited after neutralizing IL-8. Through investigating the interaction of airway epithelium and smooth muscle, our findings implicate that YKL-40 may be involved in the inflammation of asthma by induction of IL-8 from epithelium, subsequently contributing to BSMC proliferation and migration. Moreover, inhibition of IL-8 signaling is a potential therapeutic target for YKL-40-induced inflammation and remodeling of asthma.

Topics: Adipokines; Asthma; Bronchi; Cell Line, Transformed; Cell Movement; Cell Proliferation; Chitinase-3-Like Protein 1; Extracellular Signal-Regulated MAP Kinases; Humans; Inflammation Mediators; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Lectins; MAP Kinase Signaling System; Muscle, Smooth; NF-kappa B; Respiratory Mucosa; Signal Transduction; Up-Regulation

Deficient type I interferon-β and type III interferon-λ induction by rhinoviruses has previously been reported in mild/moderate atopic asthmatic adults. No studies have yet investigated if this occurs in severe therapy resistant asthma (STRA). Here, we show that compared with non-allergic healthy control children, bronchial epithelial cells cultured ex vivo from severe therapy resistant atopic asthmatic children have profoundly impaired interferon-β and interferon-λ mRNA and protein in response to rhinovirus (RV) and polyIC stimulation. Severe treatment resistant asthmatics also exhibited increased virus load, which negatively correlated with interferon mRNA levels. Furthermore, uninfected cells from severe therapy resistant asthmatic children showed lower levels of Toll-like receptor-3 mRNA and reduced retinoic acid inducible gene and melanoma differentiation-associated gene 5 mRNA after RV stimulation. These data expand on the original work, suggesting that the innate anti-viral response to RVs is impaired in asthmatic tissues and demonstrate that this is a feature of STRA.

Topics: Adolescent; Asthma; Child; Child, Preschool; DEAD Box Protein 58; DEAD-box RNA Helicases; Female; Gene Expression Regulation; Humans; Hypersensitivity, Immediate; Immunity, Innate; Immunoglobulin E; Interferon-beta; Interferon-gamma; Interferon-Induced Helicase, IFIH1; Interferons; Interleukin-8; Lung; Male; Picornaviridae Infections; Poly I-C; Receptors, Immunologic; Respiratory Mucosa; Rhinovirus; RNA, Messenger; Time Factors; Toll-Like Receptor 3

A subset of asthma patients suffer from glucocorticoid (GC) insensitivity. T-helper cell type 17 cells have an emerging role in GC insensitivity, although the mechanisms are still poorly understood. We investigated whether interleukin (IL)-17A induces GC insensitivity in airway epithelium by studying its effects on responsiveness of tumour necrosis factor (TNF)-α-induced IL-8 production to budesonide in human bronchial epithelial 16HBE cells. We unravelled the underlying mechanism by the use of specific pathway inhibitors, reporter and overexpression constructs and a histone deacetylase (HDAC) activity assay. We demonstrated that IL-17A-induced IL-8 production is normally sensitive to GCs, while IL-17A pre-treatment significantly reduced the sensitivity of TNF-α-induced IL-8 production to budesonide. IL-17A activated the p38, extracellular signal-related kinase (ERK) and phosphoinositide-3-kinase (PI3K) pathways, and the latter appeared to be involved in IL-17A-induced GC insensitivity. Furthermore, IL-17A reduced HDAC activity, and overexpression of HDAC2 reversed IL-17A-induced GC insensitivity. In contrast, IL-17A did not affect budesonide-induced transcriptional activity of the GC receptor, suggesting that IL-17A does not impair the actions of the ligated GC receptor. In conclusion, we have shown for the first time that IL-17A induces GC insensitivity in airway epithelium, which is probably mediated by PI3K activation and subsequent reduction of HDAC2 activity. Thus, blockade of IL-17A or downstream signalling molecule PI3K may offer new strategies for therapeutic intervention in GC-insensitive asthma.

Topics: Asthma; Cells, Cultured; Drug Resistance; Extracellular Signal-Regulated MAP Kinases; Glucocorticoids; Histone Deacetylase 2; Humans; Interleukin-17; Interleukin-8; p38 Mitogen-Activated Protein Kinases; Phosphatidylinositol 3-Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; Receptors, Glucocorticoid; Respiratory Mucosa; Signal Transduction; Th17 Cells; Transcription, Genetic; Tumor Necrosis Factor-alpha

Spleen tyrosine kinase (Syk) is an immunoregulatory tyrosine kinase that was identified originally in leukocytes. It is a key regulator of innate immunity as well as hematopoietic cell differentiation and proliferation. A role for Syk in regulating normal cellular functions in nonhematopoietic cells is increasingly recognized. We have shown previously robust Syk expression in airway epithelium, where it regulates the early inflammatory response to human rhinovirus (HRV) infections, and HRV cell entry by clathrin-mediated endocytosis. To test the hypothesis that Syk plays a role in modulating airway epithelial cell proliferation, migration, and production of vascular endothelial growth factor and interleukin-8, we studied the BEAS-2B human bronchial epithelial cell line and primary human airway epithelia from normal and asthmatic donors using Syk-specific pharmacologic inhibitors and small interfering RNA. Using an in vitro "wounding" model, we demonstrated significant impairment of "wound" closure after treatment with the Syk inhibitors N4-(2,2-dimethyl-3-oxo-4H-pyrid[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (R406) and 2-[7-(3,4-dimethoxyphenyl)-imidazo[1,2-c]pyrimidin-5-ylamino]-nicotinamide dihydrochloride (BAY61-3606), overexpression of the kinase-inactive Syk(K396R) mutant, and Syk knockdown by small interfering RNA. HRV infection also impaired wound healing, an effect that was partly Syk-dependent because wound healing was impaired further when HRV infection occurred in the presence of Syk inhibition. Further investigation of potential regulatory mechanisms revealed that inhibition of Syk suppressed HRV-induced vascular endothelial growth factor expression while promoting the activation of caspase-3, a mediator of epithelial cell apoptosis. Together, these results indicate that Syk plays a role in promoting epithelial cell proliferation and migration, while mitigating the effects of apoptosis.

Topics: Apoptosis; Asthma; Caspase 3; Cell Line; Cell Movement; Cell Proliferation; Cells, Cultured; Epithelial Cells; Gene Expression; Interleukin-8; Intracellular Signaling Peptides and Proteins; Protein Kinase Inhibitors; Protein-Tyrosine Kinases; Respiratory Mucosa; Rhinovirus; RNA, Small Interfering; Syk Kinase; Vascular Endothelial Growth Factor A; Wound Healing

Blood tests are needed to identify steroid-resistant (SR) asthmatic patients early so that they can be managed with alternative anti-inflammatory therapy.. We sought to assess the usefulness of peripheral blood to predict steroid response in asthmatic patients.. Nineteen asthmatic patients with FEV(1) of less than 80% of predicted value were classified as SR or steroid sensitive (SS) based on change in lung FEV(1) percentage after 7 days of oral prednisone. Blood was collected at baseline (visit 1) and 30 days after prednisone administration (visit 3). PBMCs were cultured for 4 hours with or without 10(-7) mol/L dexamethasone, and cellular response to dexamethasone was determined by using real-time PCR based on expression analysis of steroid-regulated genes. Suppression of PHA-induced T-cell proliferation by dexamethasone was assessed.. Prednisone significantly improved FEV(1) percentages in SS asthmatic patients (mean ± SE: 17.5% ± 2.4%) but not SR asthmatic patients (0.8% ± 2.0%, P < .001). Before prednisone treatment, mitogen-induced kinase phosphatase 1 (P = .01) and IL-8 mRNA (P < .05) levels were significantly higher in PBMCs from SR asthmatic patients. TNF-α (P < .05) and IL-8 fold suppression by dexamethasone (P < .05) were significantly reduced in PBMCs from SR asthmatic patients. The expression of glucocorticoid receptor (GCR) β, but not GCR-α, was significantly increased in PBMCs of SR asthmatic patients (P = .01). The dexamethasone inhibitory concentration of 50% for PBMC proliferation was significantly higher for SR asthmatic patients (P < .05). These markers no longer differed between groups in PBMCs 30 days after prednisone administration. The composite score of assays at baseline before prednisone was significantly different between SR and SS asthmatic patients (P < .001).. PBMCs from SR asthmatic patients have higher baseline mitogen-induced kinase phosphatase 1, IL-8, and GCR-β mRNA levels; have a lower GCR-α/GCR-β mRNA ratio; are less responsive to suppression of TNF-α and IL-8 by dexamethasone; and require more dexamethasone to suppress T-cell proliferation compared with SS asthmatic patients.

Topics: Adrenal Cortex Hormones; Adult; Asthma; Cell Proliferation; Cells, Cultured; Dexamethasone; Drug Resistance; Dual Specificity Phosphatase 1; Female; Humans; Interleukin-8; Leukocytes, Mononuclear; Male; Prednisone; Prognosis; Receptors, Glucocorticoid; Respiratory Function Tests; T-Lymphocytes

Patients with chronic obstructive pulmonary disease (COPD) show a poor response to corticosteroids, which has been linked to oxidative stress. Here we show that the long-acting β(2) -agonist formoterol (FM) reversed corticosteroid insensitivity under oxidative stress via inhibition of phosphoinositide-3-kinase (PI3K) signalling.. Responsiveness to corticosteroids dexamethasone (Dex), budesonide (Bud) and fluticasone propionate (FP) was determined, as IC(50) values on TNF-α-induced interleukin 8 release, in U937 monocytic cell line treated with hydrogen peroxide (H(2) O(2) ) or peripheral blood mononuclear cells (PBMCs) from patients with COPD or severe asthma.. PBMCs from severe asthma and COPD were less sensitive to Dex compared with those from healthy subjects. Both FM (10(-9)  M) and salmeterol (SM, 10(-8)  M) reversed Dex insensitivity in severe asthma, but only FM restored Dex sensitivity in COPD. Although H(2) O(2) exposure decreased steroid sensitivity in U937 cells, FM restored responsiveness to Bud and FP while the effects of SM were weaker. Additionally, FM, but not SM, partially inhibited H(2) O(2) -induced PI3Kδ-dependent (PKB) phosphorylation. H(2) O(2) decreased SM-induced cAMP production in U937 cells, but did not significantly affect the response to FM. The reduction of SM effects by H(2) O(2) was reversed by pretreatment with LY294002, a PI3K inhibitor, or IC87114, a PI3Kδ inhibitor.. FM reversed oxidative stress-induced corticosteroid insensitivity and decreased β(2) adrenoceptor-dependent cAMP production via inhibition of PI3Kδ signalling. FM will be more effective than SM, when combined with corticosteroids, for the treatment of respiratory diseases under conditions of high oxidative stress, such as in COPD.

Topics: 1-Phosphatidylinositol 4-Kinase; Adenine; Adrenal Cortex Hormones; Adrenergic beta-2 Receptor Agonists; Adult; Aged; Albuterol; Androstadienes; Asthma; Budesonide; Chromones; Dexamethasone; Drug Resistance; Ethanolamines; Female; Fluticasone; Formoterol Fumarate; Humans; Hydrogen Peroxide; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Morpholines; Oxidative Stress; Pulmonary Disease, Chronic Obstructive; Quinazolines; Receptors, Adrenergic, beta-2; Salmeterol Xinafoate; U937 Cells; Young Adult

One of the suggested health outcomes of PCB exposure is childhood asthma.. This study was conducted to find health relevant biomarkers providing the molecular epidemiological evidence for the positive relationship between exposure to PCBs and childhood asthma.. Blood samples from fifteen asthmatic children as well as an equal number of non-asthmatic children (average 2 year old) were collected, and were analyzed for PCBs and their select marker expression by using qRT-PCR.. Among biomarkers examined IL-8 expression was significantly correlated to serum levels of PCB #163+164 (P=0.022), #170 (P=0.046), #177 (P=0.022), #178 (P=0.022) and #180+193 (P=0.046) in a dose-dependent manner, which was found only among asthmatic children. In contrast, COX-2 correlations to individual congener levels were recognized only among control subjects, not among asthmatic subjects.. Serum concentrations of PCB#163+164, #170, #177, #178 and #180+193 correlate significantly with IL-8 mRNA expressions among asthmatic children.

Topics: Asthma; Biomarkers; Child, Preschool; Cyclooxygenase 2; Environmental Exposure; Environmental Pollutants; Female; Gene Expression Regulation; Humans; Immunoglobulin E; Infant; Infant, Newborn; Interleukin-8; Japan; Male; Polychlorinated Biphenyls; RNA, Messenger

The role of systemic inflammation in asthma is unclear. The aim of this study was to compare systemic inflammation in subjects with stable asthma, categorized by airway inflammatory phenotype, with healthy control subjects.. Adults with stable asthma (n = 152) and healthy control subjects (n = 83) underwent hypertonic saline challenge and sputum induction. Differential leukocyte counts were performed on selected sputum. Plasma high-sensitivity C-reactive protein (CRP), IL-6, and tumor necrosis factor-α levels and sputum IL-8 and neutrophil elastase levels were determined by enzyme-linked immunosorbent assay. Sputum IL-8 receptor α (IL-8RA) and IL-8 receptor β (IL-8RB) messenger RNA expression were determined by real-time polymerase chain reaction.. Subjects with asthma were classified as having nonneutrophilic asthma or neutrophilic asthma. The asthma (neutrophilic) group had increased systemic inflammation compared with the asthma (nonneutrophilic) and healthy control groups, with median (interquartile range) CRP levels of 5.0 (1.6-9.2), 1.8 (0.9-5.3), and 1.8 (0.8-4.1) mg/L (P = .011), respectively, and IL-6 levels of 2.1 (1.5-3.1), 1.4 (1.0-2.1), and 1.1 (0.8-1.5) pg/mL (P < .001), respectively. The proportion of subjects with elevated CRP and IL-6 levels was also higher in the asthma (neutrophilic) group. Sputum IL-8 and neutrophil elastase protein and IL-8RA and IL-8RB gene expression were significantly increased in the asthma (neutrophilic) group. In multiple regression analysis of subjects with asthma, sex, BMI, statin use, and percent sputum neutrophils were significant predictors of log(10)CRP. Sex, BMI, and %FEV(1) were significant predictors of log(10)IL-6.. Systemic inflammation is increased in patients with asthma with neutrophilic airway inflammation and associated with worse clinical outcomes. Systemic inflammation may contribute to the pathophysiology of neutrophilic asthma.

Topics: Adult; Aged; Asthma; Biomarkers; C-Reactive Protein; Case-Control Studies; Female; Humans; Inflammation; Interleukin-6; Interleukin-8; Leukocyte Elastase; Male; Middle Aged; Neutrophils; Phenotype; Sputum; Tumor Necrosis Factor-alpha

Asthma is a chronic inflammatory disease in which airway epithelial cells are the first line of defense against exposure of the airway to infectious agents. Src homology protein (SHP)-1, a protein tyrosine phosphatase, is a negative regulator of signaling pathways that are critical to the development of asthma and host defense. We hypothesize that SHP-1 function is defective in asthma, contributing to the increased inflammatory response induced by Mycoplasma pneumoniae, a pathogen known to exacerbate asthma. M. pneumoniae significantly activated SHP-1 in airway epithelial cells collected from nonasthmatic subjects by bronchoscopy with airway brushing but not in cells from asthmatic subjects. In asthmatic airway epithelial cells, M. pneumoniae induced significant PI3K/Akt phosphorylation, NF-κB activation, and IL-8 production compared with nonasthmatic cells, which were reversed by SHP-1 overexpression. Conversely, SHP-1 knockdown significantly increased IL-8 production and PI3K/Akt and NF-κB activation in the setting of M. pneumoniae infection in nonasthmatic cells, but it did not exacerbate these three parameters already activated in asthmatic cells. Thus, SHP-1 plays a critical role in abrogating M. pneumoniae-induced IL-8 production in nonasthmatic airway epithelial cells through inhibition of PI3K/Akt and NF-κB activity, but it is defective in asthma, resulting in an enhanced inflammatory response to infection.

Topics: Adult; Asthma; Bronchoalveolar Lavage Fluid; Cell Nucleus; Cells, Cultured; Epithelial Cells; Female; Humans; In Vitro Techniques; Inflammation; Interleukin-8; Male; Mycoplasma pneumoniae; NF-kappa B; Phosphatidylinositol 3-Kinases; Phosphorylation; Protein Processing, Post-Translational; Protein Tyrosine Phosphatase, Non-Receptor Type 6; Proto-Oncogene Proteins c-akt; RNA Interference; RNA, Small Interfering; Transcription, Genetic; Young Adult

The biologic mechanisms involved in airway inflammatory response to air pollution are not clearly understood. The authors conducted a longitudinal study to investigate whether exposure to ambient air pollutants affected inflammatory cells and mediators from nasal lavage in schoolchildren. Study participants were 100 elementary and middle-school students in New Taipei City, Taiwan. A structured respiratory health questionnaire was administered in September 2007, followed by monthly measurement of nasal inflammation from October 2007 to November 2009. During the study period, daily concentrations of air pollutants were obtained from the Environmental Protection Administration monitoring station and the Aerosol Supersite. Mixed-effects models were applied to examine the association between air pollution and nasal inflammatory cells and mediators, including percentages of neutrophils, eosinophils, and monocytes in lavaged cells and interleukin-8. A total of 824 measurements were obtained from 100 participants over a period of 10 months. The level of particulate matter with an aerodynamic diameter of 2.5 μm or less (PM(2.5)) was found to be associated with percentage of neutrophils (β = 3.45%, 95% confidence interval: 0.89, 6.01) and interleukin-8 level (β = 29.98 pg/mL, 95% confidence interval: 3.26, 56.69) in the nasal lavage on the day of exposure. In this longitudinal cohort study of schoolchildren, results indicated that exposure to PM(2.5) might induce nasal inflammation.

Topics: Air Pollution; Asthma; Child; Eosinophils; Female; Follow-Up Studies; Health Surveys; Humans; Inflammation; Inhalation Exposure; Interleukin-8; Longitudinal Studies; Male; Models, Statistical; Monocytes; Nasal Lavage Fluid; Neutrophils; Particulate Matter; Rhinitis; Surveys and Questionnaires; Taiwan

Structural changes to the airways are features of severe asthma. The bronchial epithelium facilitates this remodeling process. Learning about the changes that develop in the airway epithelium could improve our understanding of asthma pathogenesis and lead to new therapeutic approaches.. We sought to determine the feasibility and relevance of air-liquid interface cultures of bronchial epithelium derived from endobronchial biopsy specimens of patients with different severities of asthma for studying the airway epithelium.. Human bronchial epithelial cells derived from endobronchial biopsy specimens of patients with mild and severe asthma were maintained in culture for 21 days in an air-liquid interface to reproduce a fully differentiated airway epithelium. Initially, features of remodeling that included epithelial and subepithelial layers, as well as mucus production, were assessed in paraffin-embedded endobronchial biopsy specimens to evaluate morphologic characteristics of asthmatic patients' epithelia. Ex vivo differentiated epithelia were then analyzed for morphology and function based on ultrastructural analysis, IL-8 release, lipoxin A(4) generation, mucin production, and lipoxygenase gene expression.. Morphologic and inflammatory imbalances initially observed in endobronchial biopsy specimens obtained from patients with severe or mild asthma persisted in the air-liquid interface reconstituted epithelium throughout the differentiation process to 21 days. Epithelium from patients with severe asthma produced greater levels of mucin, released more IL-8, and produced lower levels of lipoxin A(4) than that from patients with mild asthma. Expression of 15-lipoxygenase 2 was increased in epithelium from patients with severe asthma, whereas expression levels of MUC5AC, MUC5B, 5-lipoxygenase, and 15-lipoxygeanse 1 were similar to those of patients with mild asthma.. Ex vivo cultures of fully differentiated bronchial epithelium from endobronchial biopsy specimens maintain inherent phenotypic differences specifically related to the severity of asthma.

Topics: Adult; Aged; Airway Remodeling; Asthma; Bronchi; Cell Culture Techniques; Cell Differentiation; Cells, Cultured; Disease Progression; Feasibility Studies; Female; Humans; Interleukin-8; Lipoxins; Lipoxygenase; Male; Middle Aged; Mucins; Respiratory Mucosa; Young Adult

In Taiwan, 57.5% of asthmatic patients are allergic to cockroaches, which are a major indoor allergen for immunoglobulin E (IgE)-mediated respiratory diseases.. To determine whether sensitization to different cockroach allergenic components correlates with different clinical manifestations and severities.. The complementary DNAs (cDNAs) encoding for Per a 1 through 7 and Per a 9 were generated by reverse transcription polymerase chain reaction and cloned into the Escherichia coli expression system. Sixty-four subjects were divided into 3 groups based on the clinical severity of their allergic reaction: those with persistent asthma and rhinitis (AS), those with allergic rhinitis only (AR), and the nonallergic controls (NA). Serum levels of interleukin-8 (IL-8), monocyte chemotactic protein-1 (MCP-1), chemokine (C-C motif) ligand 20 (CCL-20), and granulocyte macrophage colony-stimulating factor (GM-CSF) were measured, and the binding frequencies to each recombinant allergen were examined.. Serum levels of IL-8, MCP-1, and CCL-20 were significantly higher in the AS group than in the AR and NA groups. The numbers of IgE-binding allergens did not correlate with the clinical severity of airway allergy to cockroaches. However, 81% in the AS group had IgE-binding activity to Per a 2, which was significantly higher than that of the AR group (45%, P < .05). In contrast, 80% of AR patients had IgE-binding activity to Per a 9 compared with only 28.5% of AS patients (P < .01).. Allergens from American cockroaches do not have equal importance in terms of pathogenicity. Sensitization to Per a 2 correlates with more severe airway allergy and elevated proinflammatory chemokines. This may help in selecting target allergens for component resolved diagnosis and immunotherapeutic agents.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Allergens; Animals; Asthma; Chemokine CCL2; Chemokine CCL20; Child; Cloning, Molecular; Disease Progression; Escherichia coli; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunization; Immunoglobulin E; Insect Proteins; Interleukin-8; Male; Middle Aged; Periplaneta; Recombinant Proteins; Respiratory Hypersensitivity; Taiwan

To study the regulatory role of bacterial lipopolysaccharide (LPS ) in the development of bronchial asthma by examining the effects of LPS on serum IL-4, serum IL-8 and pulmonary vascular endothelial growth factor (VEGF) expression in mice with asthma.. Twenty-seven BALB/c mice were randomly assigned into control, asthma and LPS-treated asthma groups (n=9 each). Serum IL-4 and IL-8 concentrations were measured using ELISA. VEGF expression in lung tissues was examined using the immunohistochemical method.. Serum IL-4 and IL-8 concentrations in the asthma group were significantly higher than in the control group (P<0.05). LPS treatment significantly decreased serum IL-4 and IL-8 concentrations compared with the asthma group (P<0.05), although levels were significantly higher than in the control group (P<0.05). Airway VEGF expression in the asthma group was significantly higher than in the control group (P<0.05). LPS treatment significantly decreased airway VEGF expression compared with the asthma group (P<0.05), although concentrations remained higher than in the control group (P<0.05).. LPS can decrease serum IL-4, serum IL-8 and pulmonary VEGF expression in mice with asthma, and thus can possibly reduce both airway inflammation and airway vascular remodeling.

Topics: Animals; Asthma; Female; Interleukin-4; Interleukin-8; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Vascular Endothelial Growth Factor A

Recent investigations suggest that neutrophils may play an important role in the late-phase allergen-induced inflammation in allergic airway diseases. The aim of this study was to evaluate neutrophil chemotaxis, phagocytic activity, and reactive oxygen species (ROS) production in patients with allergic rhinitis and asthma challenged with inhaled Dermatophagoides pteronyssinus. Eighteen patients with allergic rhinitis and 14 with allergic asthma, all sensitized to D. pteronyssinus, as well as 15 healthy individuals underwent bronchial challenge with D. pteronyssinus. Peripheral blood collection and neutrophil isolation were performed 24 h before as well as 7 and 24 h after bronchial challenge. Neutrophils chemotaxis, phagocytic activity, and ROS production were analyzed by flow cytometer. Neutrophil chemotaxis and ROS production were increased, while phagocytic activity was decreased 24 h before challenge in patient groups compared with healthy individuals. After challenge, neutrophil chemotaxis and phagocytic activity increased after 7 and 24 h, when ROS production, only after 24 h. Bronchial allergen challenge had no influence for neutrophils activity in healthy subjects. Activated chemotaxis, phagocytic activity, and ROS production of peripheral blood neutrophils after challenge with D. pteronyssinus reflect an enhanced systemic inflammation in allergic rhinitis and asthma patients with induced late-phase airway inflammation.

Topics: Adult; Animals; Asthma; Chemotaxis, Leukocyte; Dermatophagoides pteronyssinus; Female; Humans; Interleukin-8; Male; Neutrophils; Phagocytosis; Reactive Oxygen Species; Respiratory Hypersensitivity; Rhinitis, Allergic, Perennial

Moxifloxacin (MXF) has been shown to possess immunomodulatory properties in addition to its antimicrobial effects. We investigated the effects of MXF on cytokine secretion and signal transduction mechanisms in naive control and allergen-exposed airway smooth muscle cell (ASMC) stimulated with tumour necrosis factor (TNF)-α.. An animal model was established. ASMC was derived from rat airway tissue and cultured in vitro, then incubated with 10 ng/mL of TNF-α. Interleukin (IL)-8 and eotaxin secretion were measured by enzyme-linked immunosorbent assay, and activation of extracellular-signal-regulated kinase (ERK)1/2 and nuclear factor (NF)-κB p65 was measured by western blotting, with or without the addition of MXF (20 µg/mL) and/or dexamethasone (DXM) (10(-6)  M).. Baseline IL-8 and eotaxin secretion did not differ between control and allergen-exposed cells. Stimulation with TNF-α increased IL-8 and eotaxin secretion, with increased IL-8 secretion by allergen-exposed compared with naive control ASMC, post-TNF-α stimulation (P = 0.001). Baseline phosphorylation of ERK1/2 (p-ERK1/2) and NF-κB p65 was higher in allergen-exposed than in control ASMC. TNF-α increased p-ERK1/2 and NF-κB p65 levels, with higher levels in allergen-exposed ASMC, post-TNF-α stimulation (P < 0.001). MXF and the combination of MXF with DXM suppressed the secretion of IL-8 and eotaxin, but DXM alone did not affect IL-8, post-TNF-α stimulation (P > 0.05). MXF, DXM and the combination of MXF with DXM inhibited TNF-α-stimulated p-ERK1/2 and NF-κB p65 levels by 34, 40 and 62%, and 33, 38 and 64%, respectively.. MXF suppressed the secretion of pro-inflammatory cytokines by allergen-exposed rat ASMC, partly by inhibiting NF-κB and ERK activation. DXM may have additional or synergistic effects with MXF.

Topics: Allergens; Animals; Anti-Infective Agents; Anti-Inflammatory Agents; Asthma; Aza Compounds; Cells, Cultured; Chemokine CCL11; Dexamethasone; Disease Models, Animal; Extracellular Signal-Regulated MAP Kinases; Fluoroquinolones; Interleukin-8; Moxifloxacin; Myocytes, Smooth Muscle; NF-kappa B; Quinolines; Rats; Tumor Necrosis Factor-alpha

Heaves, an obstructive neutrophilic airway inflammation of horses, is triggered by dust components such as endotoxin and has similarities to human asthma. Pulmonary intravascular macrophages (PIMs) increase horses' sensitivity to endotoxin-induced lung inflammation; however, their role in an airborne pathology remains unknown. Therefore, we investigated the role of PIMs in the development of heaves in horses. Clinical and inflammatory responses were evaluated following induction of heaves by moldy hay exposure and PIM depletion with gadolinium chloride (GC). Mares (N = 9) were exposed to four treatments: alfalfa cubes (Cb), alfalfa cubes + GC (Cb-GC), moldy hay (MH), and moldy hay + GC (MH-GC). Clinical scores and neutrophil concentrations in bronchoalveolar lavage (BAL) fluid were higher when mares received MH compared with MH-GC. BAL cells from MH-GC-treated mares had significantly lower IL-8 and TLR4 mRNA expression compared with MH-treated mares. In vitro LPS challenge significantly increased IL-8 but not TLR4 mRNA expression in BAL cells recovered from horses fed with MH, but not from the MH-GC treatment. In summary, PIM depletion attenuated clinical scores, reduced the alveolar migration of neutrophils, and decreased the expression of proinflammatory molecules in BAL cells of heaves horses, suggesting a proinflammatory role of PIMs in the development of airborne pathology.

Topics: Airway Obstruction; Animals; Asthma; Blotting, Western; Bronchoalveolar Lavage; Dust; Endotoxins; Female; Horse Diseases; Horses; Humans; Interleukin-8; Lipopolysaccharides; Macrophages, Alveolar; Neutrophils; Pneumonia; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Exposure to particulate matter (PM) is associated with adverse pulmonary effects, including induction and exacerbation of asthma. Recently arginase was shown to play an important role in the pathogenesis of asthma. In this study, it was postulated that PM exposure might induce arginase. Human bronchial epithelial cells (HBEC) obtained from normal individuals by endobronchial brushings cultured on an air-liquid interface were incubated with fine Chapel Hill particles (PM₂.₅, 100 μg/ml) for up to 72 h. Arginase activity, protein expression, and mRNA of arginase I and arginase II were measured. PM₂.₅ increased arginase activity in a time-dependent manner. The rise was primarily due to upregulation of arginase II. PD153035 (10 μM), an epidermal growth factor (EGF) receptor antagonist, attenuated the PM₂.₅-induced elevation in arginase activity and arginase II expression. Treatment of HBEC with human EGF increased arginase activity and arginase II expression. Pretreatment with catalase (200 U/ml), superoxide dismutase (100 U/ml), or apocynin (5 μg/ml), an NAD(P)H oxidase inhibitor, did not markedly affect arginase II expression. Treatment of HBEC with arginase II siRNA inhibited the expression of arginase II by 60% and increased IL-8 release induced by PM₂.₅. These results indicate that PM exposure upregulates arginase II activity and expression in human bronchial epithelial cells, in part via EGF-dependent mechanisms independent of oxidative stress. The elevated arginase II activity and expression may be a mechanism underlying adverse effects induced by PM exposure in asthma patients.

Topics: Air Pollutants; Arginase; Asthma; Bronchi; Cells, Cultured; Enzyme Induction; Epidermal Growth Factor; ErbB Receptors; Humans; Interleukin-8; Isoenzymes; North Carolina; Particulate Matter; Recombinant Proteins; Respiratory Mucosa; RNA Interference; RNA, Messenger; RNA, Small Interfering; Time Factors

To explore the effect of Toll-like receptor 4 (TLR4) activation on the migration of asthmatic airway smooth muscle cell (ASMCs) induced by airway epithelial cells.. Primary ASMCs were cultured by the method of cell digestion. Cell culture supernatant of RTE cells were collected by TNF-alpha stimulation of epithelial cells. Detected the IL-8 and RANTES levels in the supernatant. The transmembrane migration of asthmatic ASMCs were detected by Modified Boyden chemotaxis chamber. The effect of TLR4 on the migration of asthmatic ASMCs induced by epithelial cells with TLR4 antibody drugs as a tool.. The levels of IL-8 and RANTES in the supernatant of TNF-alpha groups were significantly increased, and that in the 20 ng/ml group was significantly higher than other groups (P < 0.01). The transmembrane migration of asthmatic ASMCs groups was increased than that of control group. The transmembrane migration of asthmatic ASMCs from asthma group and TNF-alpha + TLR4 antibody group was significantly decreased compared with that in TNF-alpha group (P < 0.01). The migration of asthma ASMCs from TNF-alpha + TLR4 antibody group was increased than that of asthma group (P < 0.05).. TLR4 in the surface of asthmatic ASMCs may be activated by cytokines secreted by the airway epithelial cells and enhance the transmembrane migration of asthmatic ASMCs induced by airway epithelial cells so that it plays a role in airway remodeling of asthma.

Topics: Animals; Asthma; Cell Movement; Cells, Cultured; Chemokine CCL5; Epithelial Cells; Interleukin-8; Myocytes, Smooth Muscle; Rats; Rats, Sprague-Dawley; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Severe asthma accounts for a small number of asthmatics but represents a disproportionate cost to health care systems. The underlying mechanism in severe asthma remains unknown but several mechanisms are likely to be involved because of a very heterogeneous profile. We investigated the effects of a p38MAPK inhibitor in corticosteroid sensitivity in peripheral blood mononuclear cells (PBMCs) from severe asthmatics and the profile of its responders.. Corticosteroid sensitivity was determined by measuring dexamethasone inhibition of CD3/28 and TNF-α induced IL-8 production in PBMCs by using ELISA. PBMCs from severe asthmatics were relatively less sensitive to dexamethasone (Dex) as compared to those of non-severe asthmatics and healthy volunteers. The IC(50) values of Dex negatively correlated with decreased glucocorticoid receptor (GR) nuclear translocation assessed using immunocytochemistry (r = -0.65; p<0.0005) and with decreased FEV(1) (% predicted) (r = 0.6; p<0.0005). A p38α/β inhibitor (SB203580) restored Dex-sensitivity in a subpopulation of severe asthma that was characterized by a defective GR nuclear translocation, clinically by lower FEV(1) and higher use of oral prednisolone. We also found that SB203580 partially inhibited GR phosphorylation at serine 226, resulting in increased GR nuclear translocation in IL-2/IL-4 treated corticosteroid insensitive U937s.. p38MAPKα/β is involved in defective GR nuclear translocation due to phosphorylation at Ser226 and this will be a useful biomarker to identify responders to p38MAPKα/β inhibitor in the future.

Topics: Adrenal Cortex Hormones; Adult; Asthma; Cell Nucleus; Female; Humans; Imidazoles; Interleukin-8; Leukocytes, Mononuclear; Male; Models, Biological; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Phosphoserine; Protein Kinase Inhibitors; Protein Transport; Pyridines; Receptors, Glucocorticoid; Severity of Illness Index; U937 Cells

Allergic asthma is showed an increase in Th2-cytokine and IgE levels and an accumulation activation of Th2 cells, eosinophils and mast cells. However, recent studies focused on cell-based mechanisms for the pathogenesis of allergic asthma. Objectives. In this study, we compare the anti-IgE treatment modality in the dynamics of immune system cytokine levels in severe persistent asthma (SPA) patients who had no other any allergic disease, newly diagnosed allergic asthma patients and healthy volunteers.. The study population consisted of 14 SPA patients, 14 newly diagnosed allergic asthma patients and 14 healthy volunteers included as controls. Cytokine levels were measured. Total and specific IgE levels of anti-IgE monoclonal antibody treated patients, serum high-sensitivity C-reactive protein (hsCRP) levels, FEV1/FVC rates and asthma control test (ACT) were measured for the clinical follow-up.. We observed that SPA patients presented increasing levels of IL-8, IL-10, TGF-β and GCSF during the anti-IgE treatment in period of sampling times at 4 months and 18 months. However this increase was not correlated neither with serum hsCRP levels nor FEV1/FVC rates.. Our study gives a different perspective for the SPA and anti-IgE immunotherapy efficacy at the cell cytokine-linked step.

Topics: Adolescent; Adult; Antibodies, Anti-Idiotypic; Asthma; C-Reactive Protein; Cytokines; Female; Forced Expiratory Volume; Granulocyte Colony-Stimulating Factor; Humans; Interleukin-10; Interleukin-8; Male; Middle Aged; Transforming Growth Factor beta

Airway cellular dysfunction is a differentiating feature of severe asthma in children that may be related to an imbalance of the antioxidant, glutathione (GSH). We hypothesized that oxidation of GSH to glutathione disulfide (GSSG) in the epithelial lining fluid (ELF) of children with severe asthma would contribute to altered airway macrophage (AM) GSH homeostasis and AM cellular dysfunction. Bronchoalveolar lavage (BAL) was performed in 64 asthmatic children (severe asthma, n = 43). GSH, GSSG, markers of lipid peroxidation and DNA oxidation, and IL-8 were quantified in the BAL supernatant. GSH, GSSG, activities of histone deacetylase (HDAC) and histone acetyltransferase, apoptosis, and phagocytosis were assessed in isolated AMs. Children with severe asthma had increased GSSG, lipid peroxidation, byproducts of DNA oxidation, and inflammation in the ELF. This imbalance of GSH homeostasis was also noted intracellularly within the AMs and was associated with decreased HDAC activities, increased apoptosis, and impaired phagocytosis. In vitro GSH supplementation inhibited apoptosis and rescued phagocytosis in children with severe asthma. Severe asthma in children is characterized by altered airway and intracellular AM GSH homeostasis that translates to impaired AM function. Interventions to restore airway GSH homeostasis may be warranted in children with severe asthma.

Topics: Adolescent; Apoptosis; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Bronchoscopy; Child; DNA Damage; Female; Glutathione; Glutathione Disulfide; Histone Acetyltransferases; Histone Deacetylases; Humans; Interleukin-8; Lipid Peroxidation; Macrophages, Alveolar; Male; Oxidation-Reduction; Oxidative Stress; Phagocytosis; Severity of Illness Index; Spirometry

The increase in airway smooth muscle (ASM) mass is a major structural change in asthma. This increase has been attributed to ASM cell (ASMC) hyperplasia and hypertrophy. The distance between ASMC and the epithelium is reduced, suggesting migration of smooth muscle cells toward the epithelium. Recent studies have suggested a role of chemokines in ASMC migration toward the epithelium; however, chemokines have other biological effects. The objective of the current study is to test the hypothesis that chemokines (eotaxin, RANTES, IL-8, and MIP-1α) can directly influence ASMC mass by increasing the rate of proliferation or enhancing the survival of these cells. Human ASMCs were exposed to different concentrations of eotaxin, RANTES, IL-8, or MIP-1α. To test for proliferation, matched control and stimulated ASMC were pulsed with [(3)H]thymidine, or ASMCs were stained with BrdU and then analyzed with flow cytometry. Apoptosis was measured using Annexin V staining and flow cytometry. Expression of phosphorylated p42/p44 and MAPKs was assessed by Western blot. In a concentration-dependent manner, chemokines including eotaxin, RANTES, IL-8, and MIP-1α increased ASMC's [(3)H]thymidine incorporation and DNA synthesis. IL-8, eotaxin, and MIP-1α decreased the rate of apoptosis of ASMCs compared with the matched controls. A significant increase in phosphorylated p42/p44 MAPKs was seen after treating ASMCs with RANTES and eotaxin. Moreover, inhibition of p42/p44 MAPK phosphorylation reduced the level of chemokine-induced ASM proliferation. We conclude that chemokines might contribute to airway remodeling seen in asthma by enhancing the number and survival of ASMCs.

Topics: Airway Remodeling; Apoptosis; Asthma; Bronchi; Cell Proliferation; Cell Survival; Cells, Cultured; Chemokine CCL11; Chemokine CCL3; Chemokine CCL5; DNA Replication; Down-Regulation; Humans; Interleukin-8; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Myocytes, Smooth Muscle; Phosphorylation; Thymidine; Up-Regulation

Severe or refractory asthma affects 5 to 15% of all patients with asthma, but is responsible for more than half of the health burden associated with the disease. Severe asthma is characterized by a dramatic increase in smooth muscle and airway inflammation. Although glucocorticoids are the mainstay of treatment in asthma, they are unable to fully control the disease in individuals with severe asthma. We found that airway smooth muscle cells (ASMCs) from individuals with severe asthma showed elevated activities of the ERK1/ERK2 and p38 MAPK pathways despite treatment with oral and inhaled glucocorticoids, which increased the expression of DUSP1, a phosphatase shown to limit p38 MAPK activity. In ex vivo ASMCs, TNF-α but not IL-17A induced expression of the neutrophil chemoattractant CXCL8. Moreover, TNF-α led to up-regulation of the ERK1/ERK2 and p38 MAPKs pathways, with only the latter being sensitive to pretreatment with the glucocorticoid dexamethasone. In contrast to epithelial and endothelial cells, TNF-α-stimulated CXCL8 synthesis was dependent on ERK1/ERK2 but not on p38 MAPK. Moreover, suppressing ERK1/ERK2 activation prevented neutrophil recruitment by ASMCs, whereas suppressing p38 MAPK activity had no impact. Taken together, these results highlight the ERK1/ERK2 MAPK cascade as a novel and attractive target in severe asthma because the activation of this pathway is insensitive to the action of glucocorticoids and is involved in neutrophil recruitment, contributing the to inflammation seen in the disease.

Topics: Adult; Aged; Aged, 80 and over; Anti-Asthmatic Agents; Asthma; Cells, Cultured; Dual Specificity Phosphatase 1; Female; Glucocorticoids; Humans; Interleukin-8; Male; MAP Kinase Signaling System; Middle Aged; Muscle, Smooth; Myocytes, Smooth Muscle; Neutrophils; Severity of Illness Index; Tumor Necrosis Factor-alpha; Up-Regulation; Young Adult

Nuclear factor-κB (NF-κB) is a transcriptional factor of different inflammatory patterns involved in asthma and chronic obstructive pulmonary disease (COPD) that is tightly controlled by IκB kinase (IKK) complex.. We investigated the dysregulation of IKK-driven NF-κB activation in patients with asthma and COPD.. We assessed IKKα and IKKβ expression and activation, their regulation by glucocorticosteroids, and their involvement in IL-8 synthesis in PBMCs isolated from asthmatic patients, healthy smokers (HSs), patients with COPD, and control subjects. PBMCs from control subjects were stimulated with TNF-α and cigarette smoke extract in the presence or absence of fluticasone propionate (FP), L-glutathione reduced, or both, and IKK activation and IL-8 release were evaluated.. IKKα activity was higher in patients with COPD and HSs than in asthmatic patients and control subjects. IKKβ activity was higher in asthmatic patients, HSs, and patients with COPD than in control subjects. In vitro FP treatment induced inhibition of both IKKα and IKKβ activity in PBMCs from asthmatic patients, patients with COPD, and HSs, although IKKβ activity was more sensitive to FP than that of IKKα. FP reduced the IL-8 released from PBMCs of asthmatic patients, patients with COPD, and HSs, although IL-8 inhibition was higher in asthmatic patients than in patients with COPD and HSs. FP reduced IKKα and IKKβ activities in TNF-α and cigarette smoke extract-treated PBMCs, with higher levels of inhibition for IKKβ than IKKα activity. L-glutathione reduced improved the downregulatory effects of FP on IKKα and IL-8 levels.. Based on differential activation of IKKα and IKKβ, our findings suggest a different profile in the upstream regulation of the IKK-driven NF-κB system in asthmatic patients and patients with COPD. These differences in the regulation of the inflammatory process may explain, at least in part, the different pharmacologic responses in these patients.

Topics: Adult; Asthma; Bronchodilator Agents; Enzyme Activation; Female; Gene Expression Regulation; Glucocorticoids; Humans; I-kappa B Kinase; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; NF-kappa B; Pulmonary Disease, Chronic Obstructive; Smoking

Surfactant protein A (SP-A) regulates a variety of immune cell functions. We determined the ability of SP-A derived from normal and asthmatic subjects to modulate the inflammatory response elicited by Mycoplasma pneumoniae, a pathogen known to exacerbate asthma. Fourteen asthmatic and 10 normal control subjects underwent bronchoscopy with airway brushing and bronchoalveolar lavage (BAL). Total SP-A was extracted from BAL. The ratio of SP-A1 to total SP-A (SP-A1/SP-A) and the binding of total SP-A to M. pneumoniae membranes were determined. Airway epithelial cells from subjects were exposed to either normal or asthmatic SP-A before exposure to M. pneumoniae. IL-8 protein and MUC5AC mRNA were measured. Total BAL SP-A concentration did not differ between groups, but the percentage SP-A1 was significantly increased in BAL of asthmatic compared with normal subjects. SP-A1/SP-A significantly correlated with maximum binding of total SP-A to M. pneumoniae, but only in asthma. SP-A derived from asthmatic subjects did not significantly attenuate IL-8 and MUC5AC in the setting of M. pneumoniae infection compared with SP-A derived from normal subjects. We conclude that SP-A derived from asthmatic subjects does not abrogate inflammation effectively, and this dysfunction may be modulated by SP-A1/SP-A.

Topics: Adult; Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Bronchoscopy; Case-Control Studies; Cell Membrane; Cells, Cultured; Epithelial Cells; Female; HEK293 Cells; Humans; Inflammation; Interleukin-8; Male; Mucin 5AC; Mycoplasma pneumoniae; Plasmids; Polymerase Chain Reaction; Protein Binding; Pulmonary Surfactant-Associated Protein A; Recombinant Proteins; RNA, Messenger; Transfection

There is a need for reproducible and effective models of pediatric bronchial epithelium to study disease states such as asthma. We aimed to develop, characterize, and differentiate an effective, an efficient, and a reliable three-dimensional model of pediatric bronchial epithelium to test the hypothesis that children with asthma differ in their epithelial morphologic phenotype when compared with nonasthmatic children. Primary cell cultures from both asthmatic and nonasthmatic children were grown and differentiated at the air-liquid interface for 28 d. Tight junction formation, MUC5AC secretion, IL-8, IL-6, prostaglandin E2 production, and the percentage of goblet and ciliated cells in culture were assessed. Well-differentiated, multilayered, columnar epithelium containing both ciliated and goblet cells from asthmatic and nonasthmatic subjects were generated. All cultures demonstrated tight junction formation at the apical surface and exhibited mucus production and secretion. Asthmatic and nonasthmatic cultures secreted similar quantities of IL-8, IL-6, and prostaglandin E2. Cultures developed from asthmatic children contained considerably more goblet cells and fewer ciliated cells compared with those from nonasthmatic children. A well-differentiated model of pediatric epithelium has been developed that will be useful for more in vivo like study of the mechanisms at play during asthma.

Topics: Asthma; Bronchi; Child; Dinoprostone; Epithelium; Humans; Interleukin-6; Interleukin-8; Models, Biological; Mucin 5AC; Tight Junctions

Asthma is characterised into eosinophilic and non-eosinophilic phenotypes based on inflammatory cell patterns in airway secretions. Neutrophils are important in innate immunity, and are increased in the airways in non-eosinophilic asthma. The present study investigated the activity of neutrophils in asthma phenotypes. Participants with eosinophilic (n = 8) and non-eosinophilic asthma (n = 9) and healthy controls (n = 11) underwent sputum induction and blood collection. Neutrophils were isolated and cultured with or without lipopolysaccharide. Cytokines were measured by ELISA, and gene expression was analysed using a gene expression microarray and quantitative PCR. In non-eosinophilic asthma, blood neutrophils released significantly higher levels of interleukin-8 at rest. Cytokine gene expression and sputum neutrophil protein production did not differ between asthma subtypes. Microarrays demonstrated closely related expression profiles from participants with non-eosinophilic asthma that were significantly distinct from those in eosinophilic asthma. A total of 317 genes were significantly altered in resting neutrophils from participants with non-eosinophilic asthma versus eosinophilic asthma, including genes related to cell motility and regulation of apoptosis. Non-eosinophilic and eosinophilic asthma are associated with specific gene expression profiles, providing further evidence that these phenotypes of asthma involve different molecular mechanisms of disease pathogenesis at the systemic level. The mechanisms of non-eosinophilic asthma may involve enhancement of blood neutrophil chemotaxis and survival.

Topics: Adult; Aged; Asthma; Case-Control Studies; Eosinophilia; Female; Gene Expression Profiling; Humans; Interleukin-8; Male; Middle Aged; Neutrophils; Oligonucleotide Array Sequence Analysis; Sputum

Pregnancy can influence the course of maternal asthma, but the mechanisms are presently unknown. The aim of the present study was to access maternal immune cell profiles in the presence and absence of asthma and to determine the effect of pregnancy-derived factors on epithelial cell function.. Cells from the human bronchial epithelial cell line BEAS-2B were treated with plasma from pregnant or nonpregnant asthmatic and nonasthmatic subjects. Cell culture supernatants were collected after 24 h and assayed for IL-6, IL-8, eotaxin, RANTES and sICAM-1 protein using ELISA. Maternal immune cell count and peripheral blood chemotactic response to plasma from pregnant and non-pregnant asthmatic subjects were also assessed.. The presence of maternal asthma during pregnancy was associated with increased monocyte and neutrophil numbers, increased BEAS-2B cell production of IL-8 and sICAM-1 (P < 0.05) and increased chemotactic capacity relative to pregnant women without asthma.. The results of this study suggest that circulating pregnancy-related factors enhance chemotactic mediators in epithelial cells in the presence of asthma. This may be one mechanism that contributes to pregnancy-induced changes in asthma.

Topics: Adult; Asthma; Cell Line; Chemokine CCL11; Chemokine CCL5; Chemokines; Chemotaxis; Epithelial Cells; Female; Glucocorticoids; Humans; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; Pregnancy; Pregnancy Complications; Respiratory Mucosa

Potential bacterial pathogens are found in the airways in several diseases that are associated with neutrophilic inflammation. The aim of this study was to characterize subjects with stable asthma, with no symptoms of respiratory infection, to assess whether key potentially pathogenic bacteria were present in significant quantities in the airways and to correlate this with the pattern of airway inflammation and oxidative stress. Subjects with stable asthma (n = 115) and healthy controls (n = 8) underwent clinical assessment, including hypertonic saline challenge combined with sputum induction. A significant load of potentially pathogenic bacteria (> 10(6) cfu/mL) was cultured from the sputum of 17 (15%) subjects with stable asthma and was associated with higher total cell counts, proportion and number of neutrophils, sputum IL-8 and 8-isoprostane concentrations. The role of bacteria in potentiating neutrophilic asthma warrants further investigation. Therapies such as antibiotic and antioxidant treatment may be most effective in this sub-group of patients.

Topics: Adult; Aged; Asthma; Bacteria; Dinoprost; Female; Humans; Inflammation; Interleukin-8; Male; Middle Aged; Neutrophils; Oxidative Stress; Sputum

The recurrent hypoxic stress that characterizes obstructive sleep apnea (OSA) seems to play a role in the increased adherence of neutrophils to endothelial cells as well as in the resulting migration of the former to the inflamed area. Intercellular adhesion molecule 1 (ICAM-1) and interleukin (IL)-8 are markers widely used in OSA studies to investigate inflammation. The aim of this study was to measure ICAM-1 and IL-8 levels in the breath condensate and in the plasma and inflammatory cells in the induced sputum of 12 obese OSA (OO) patients, 10 nonobese OSA (NOO) patients, 10 obese non-OSA (ONO) subjects, and 8 healthy subjects (HS) using a specific enzyme immunoassay (EIA) kit. A significant increase in both plasma and exhaled IL-8 and ICAM concentrations and percentage neutrophils was observed in the induced sputum of obese OSA patients, non-obese OSA patients, and obese non-OSA subjects compared with healthy subjects. However, although these inflammatory markers were found to follow an upward trend in obese OSA patients no difference was observed in both either non-obese OSA patients and obese non-OSA subjects. Finally, a significant positive correlation was found to occur among IL-8, ICAM-1, and sputum neutrophils, as well as across the apnea-hypopnoea index (AHI), TST 90%, body mass index (BMI), and neck circumference. The data obtained confirm the occurrence of an ICAM- and IL-8-mediated neutrophilic airway inflammation in both OSA and obese patients. The degree of inflammation, which seems to worsen in cases of comorbidity (OSA and obesity), is likely to be responsible for the increased risk of developing cardiovascular events observed in these subjects, and therefore, it deserves to be elucidated even more.

Topics: Adult; Aged; Asthma; Breath Tests; Cell Count; Female; Humans; Inflammation; Intercellular Adhesion Molecule-1; Interleukin-8; Male; Middle Aged; Neutrophils; Obesity; Sleep Apnea, Obstructive; Sputum

Asthma is a chronic inflammatory condition. Inhibition of the ubiquitin-proteasome system offers promise as a anti-inflammatory strategy, being responsible for the degradation of key proteins involved in crucial cellular functions, including gene expression in inflammation (e.g. inhibitory IkappaB-alpha and the endogenous MAPK deactivator - MKP-1). As MKP-1 inhibits MAPK-mediated pro-remodeling functions in human airway smooth muscle (ASM; a pivotal immunomodulatory cell in asthma) in this study we investigate the effect of the proteasome inhibitor MG-132 on MKP-1 and evaluate the anti-inflammatory effect of MG-132 on cytokine secretion from ASM cells. Examining the time-course of induction of MKP-1 mRNA and protein by MG-132 (10microM) we show that MKP-1 mRNA was first detected at 30min, increased to significant levels by 4h, resulting in a 12.6+/-1.5-fold increase in MKP-1 mRNA expression by 24h (P<0.05). MKP-1 protein levels corroborate the mRNA results. Investigating the effect of MG-132 on secretion of the cytokine IL-6 we show that while short-term pretreatment with MG-132 (30min) partially reduced TNFalpha-induced IL-6 via inhibition of IkappaB-alpha degradation and the NF-kappaB pathway, longer-term proteasome inhibition (up to 24h) robustly upregulated MKP-1 and was temporally correlated with repression of p38-mediated IL-6 secretion from ASM cells. Moreover, utilizing a cytokine array we show that MG-132 represses the secretion of multiple cytokines implicated in asthma. Taken together, our results demonstrate that MG-132 upregulates MKP-1 and represses cytokine secretion from ASM and highlight the potential of the proteasome as a therapeutic target in asthma.

Topics: Asthma; Cysteine Proteinase Inhibitors; Cytokines; Dual Specificity Phosphatase 1; Humans; Interleukin-6; Interleukin-8; Leupeptins; Mitogen-Activated Protein Kinases; Myocytes, Smooth Muscle; Proteasome Endopeptidase Complex; Proteasome Inhibitors; Respiratory Mucosa; Signal Transduction; Tumor Necrosis Factor-alpha; Up-Regulation

Stress-elicited disruption of immunity begins in utero.. Associations among prenatal maternal stress and cord blood mononuclear cell (CBMC) cytokine responses were prospectively examined in the Urban Environment and Childhood Asthma Study (n = 557 families).. Prenatal maternal stress included financial hardship, difficult life circumstances, community violence, and neighborhood/block and housing conditions. Factor analysis produced latent variables representing three contexts: individual stressors and ecological-level strains (housing problems and neighborhood problems), which were combined to create a composite cumulative stress indicator. CBMCs were incubated with innate (lipopolysaccharide, polyinosinic-polycytidylic acid, cytosine-phosphate-guanine dinucleotides, peptidoglycan) and adaptive (tetanus, dust mite, cockroach) stimuli, respiratory syncytial virus, phytohemagglutinin, or medium alone. Cytokines were measured using multiplex ELISAs. Using linear regression, associations among increasing cumulative stress and cytokine responses were examined, adjusting for sociodemographic factors, parity, season of birth, maternal asthma and steroid use, and potential pathway variables (prenatal smoking, birth weight for gestational age).. Mothers were primarily minorities (Black [71%], Latino [19%]) with an income less than $15,000 (69%). Mothers with the highest cumulative stress were older and more likely to have asthma and deliver lower birth weight infants. Higher prenatal stress was related to increased IL-8 production after microbial (CpG, PIC, peptidoglycan) stimuli and increased tumor necrosis factor-alpha to microbial stimuli (CpG, PIC). In the adaptive panel, higher stress was associated with increased IL-13 after dust mite stimulation and reduced phytohemagglutinin-induced IFN-gamma.. Prenatal stress was associated with altered innate and adaptive immune responses in CBMCs. Stress-induced perinatal immunomodulation may impact the expression of allergic disease in these children.

Topics: Adolescent; Adult; Asthma; Black or African American; Female; Fetal Blood; Hispanic or Latino; Humans; Infant, Low Birth Weight; Infant, Newborn; Interferon-gamma; Interleukin-13; Interleukin-8; Leukocytes, Mononuclear; Male; Poverty; Pregnancy; Pregnancy Complications; Stress, Physiological; Tumor Necrosis Factor-alpha; Urban Population; Young Adult

SB203580 is the prototypical p38 MAPK inhibitor; however it cannot be used clinically due to liver toxicity. We developed a structural analogue of SB203580 - ML3403 - with equal in vitro and ex vivo p38alpha MAPK inhibition as SB203580, but with reduced activity towards liver cytochrome P450 enzymes. In addition, we developed a selective p38alpha MAPK inhibitor - CP41. The aim of this study is to compare the anti-inflammatory activity of ML3403 and CP41, with SB203580. We compare and contrast the ability of the p38 MAPK inhibitors to repress tumour necrosis factor alpha (TNFalpha)-induced interleukin 6 (IL-6) and interleukin 8 (IL-8) mRNA expression and protein secretion from airway smooth muscle cells. We also examined and compared the binding affinities of ML3403 and SB203580 to the active and inactive p38alpha MAPK. We demonstrate that ML3403 binds to both active and inactive p38 MAPK with high affinity and that it inhibits p38 MAPK-mediated airway smooth muscle synthetic function to an equivalent degree with SB203580. CP41 was not able to reduce IL-6 and IL-8 secretion in airway smooth muscle cells; a function of its higher IC(50) against p38alpha MAPK when compared to SB203580 and ML3403. We show that p38 MAPK-mediated pro-inflammatory pathways in airway smooth muscle cells can be inhibited by ML3403. The anti-inflammatory activity is equivalent to the prototypical p38 MAPK inhibitor SB203580. Our results implicate a future pharmacotherapeutic strategy towards reducing inflammation in asthma and airway remodelling.

Topics: Anti-Inflammatory Agents; Asthma; Cell Line; Gene Expression Regulation; Humans; Imidazoles; Interleukin-6; Interleukin-8; Isoenzymes; Muscle, Smooth; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Protein Kinase Inhibitors; Pyridines; Respiratory System; RNA, Messenger; Tumor Necrosis Factor-alpha

Neutrophilic inflammation plays an important role in lung tissue destruction occurring in many chronic pulmonary diseases. Neutrophils can be recruited to sites of inflammation via the action of the cytokine IL-17. In this study, we report that IL-17RA and IL-17RC mRNA expression is significantly increased in asthmatic bronchoscopic biopsies and that these receptors are not only expressed on epithelial and inflammatory cells but also on endothelial cells. IL-17 potently stimulates lung microvascular endothelial cells to produce chemoattractants (CXCL8 and derivatives of the 5-lipoxygenase pathway) that selectively drive neutrophil but not lymphocyte chemotaxis. Moreover, IL-17 promotes endothelial activation by inducing the expression of endothelial adhesion markers (E-selectin, VCAM-1, and ICAM-1) in a p38 MAPK-dependent manner. This increased expression of adhesion molecules stimulates the trans-endothelial migration of neutrophils, as well as the transmigration of HT-29 colon carcinoma cells, suggesting a further role in promoting lung metastasis. Finally, IL-17 increased neutrophil adhesion to the endothelium in vivo as determined by intravital microscopy of mice cremaster muscle. Overall, our results demonstrate that IL-17 is a potent activator of the endothelium in vivo leading to neutrophil infiltration. Therefore, preventing neutrophil recruitment by blocking the action of IL-17 on endothelial cells may prove to be highly beneficial in diseases in which neutrophilic inflammation plays a key role.

Topics: Adult; Animals; Asthma; Cells, Cultured; Endothelium, Vascular; HT29 Cells; Humans; Inflammation; Inflammation Mediators; Interleukin-17; Interleukin-8; Jurkat Cells; Male; MAP Kinase Signaling System; Mice; Neutrophil Infiltration; Neutrophils; p38 Mitogen-Activated Protein Kinases

To investigate the expressions of 15-lipoxygenase (15-LO) and 5-lipoxygenase (5-LO) in leukocytes and the changes of blood lipoxin A(4)(LXA(4)), leukotriene (LT)B(4) and LTC(4) in children with asthma, and to explore the relationship between the blood eicosanoids and one of serum high-sensitivity C-reactive protein (hsCRP), interleukin (IL)-5, IL-8 and IL-13 and IgE in children with asthma.. One hundred six asthmatic children were divided into three groups, that is, mild persistent asthmatic group, moderate persistent asthmatic group and severe persistent asthmatic group. Forty healthy children were served as controls.. The expressions of 15-LO and 5-LO mRNA in leukocytes were assessed by reverse transcription-polymerase chain reaction, and the blood LXA(4), LTB(4), LTC(4), IL-5, IL-8, and IL-13 were determined with enzyme-linked immunosorbent assay. Serum hsCRP was determined with latex-enhanced immuno-turbidimetry kits.. The leukocytic 15-LO expression and blood LXA(4) were gradually decreased, and the leukocytic 5-LO expression, blood LTB(4), LTC(4), IL-5, IL-8, IL-13, and hsCRP were gradually increased in children with asthma from mild degree to moderate and severe degree. There were positive correlations between blood LXA(4) and one of the peak expiratory flow (PEF) and forced expiratory volume in 1 sec (FEV(1)) percent-predicted values, and negative correlations between blood LTC(4) and one of the PEF and FEV(1) percent-predicted values in children with asthma. There were negative correlations between blood LXA(4) and one of the IL-5, IL-8, IL-13, and hsCRP levels, and positive correlations between one of blood LTB(4), LTC(4) and one of the IL-5, IL-8, IL-13 and hsCRP levels in children with asthma.. The reversed changes between 15-LO, its product LXA(4) and 5-LO, its products LTB(4) and LTC(4) in children with asthma from mild, moderate to severe degree were found, suggesting that insufficient generation of LXA(4) and overproduction of LTs may be the reason for the asthmatic children whose illness become more serious.

Topics: Asthma; C-Reactive Protein; Child; Child, Preschool; Female; Humans; Immunoglobulin E; Interleukin-13; Interleukin-5; Interleukin-8; Leukotrienes; Lipoxins; Male; Respiratory Function Tests

Due to their potent bronchodilator properties, beta(2)-adrenoceptor agonists are a mainstay of therapy in asthma. However, the effects of beta(2)-adrenoceptor agonists on inflammation are less clear. Accordingly, we have investigated the effects of beta(2)-adrenoceptor agonists on inflammatory mediator release.. Transcription factor activation, and both release and mRNA expression of IL-6 and IL-8 were examined by luciferase reporter assay, elisa and real-time RT-PCR in bronchial human epithelial BEAS-2B cells or primary human bronchial epithelial cells grown at an air-liquid interface.. Pre-incubation with beta(2)-adrenoceptor agonists (salbutamol, salmeterol, formoterol) augmented the release and mRNA expression of IL-6 and IL-8 induced by IL-1beta and IL-1beta plus histamine, whereas NF-kappaB-dependent transcription was significantly repressed, and AP-1-dependent transcription was unaffected. These effects were mimicked by other cAMP-elevating agents (PGE(2), forskolin). Enhancement of cytokine release by beta(2)-adrenoceptor agonists also occurred in primary bronchial epithelial cells. Addition of dexamethasone with salmeterol repressed IL-6 and IL-8 release to levels that were similar to the repression achieved in the absence of salmeterol. IL-6 release was enhanced when salmeterol was added before, concurrently or after IL-1beta plus histamine stimulation, whereas IL-8 release was only enhanced by salmeterol addition prior to stimulation.. Enhancement of IL-6 and IL-8 release may contribute to the deleterious effects of beta(2)-adrenoceptor agonists in asthma. As increased inflammatory mediator expression is prevented by the addition of glucocorticoid to the beta(2)-adrenoceptor, our data provide further mechanistic support for the use of combination therapies in asthma management.

Topics: Adrenergic beta-2 Receptor Agonists; Adrenergic beta-Agonists; Albuterol; Asthma; Bronchi; Bronchodilator Agents; Cell Line; Dexamethasone; Epithelial Cells; Ethanolamines; Formoterol Fumarate; Gene Expression Regulation; Glucocorticoids; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Salmeterol Xinafoate; Transcription, Genetic

Topics: Anti-Asthmatic Agents; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Asthma; Blotting, Western; Clinical Trials as Topic; Cyclic AMP Response Element-Binding Protein; Forced Expiratory Volume; Forkhead Transcription Factors; Humans; Immunohistochemistry; Inflammation; Interleukin-8; Omalizumab; Time

Little information is available on eosinophil activation and cytokine/chemokine responses in childhood asthma, thus we examined serum eosinophil cationic protein (ECP) and 27 types of cytokines/chemokines in acute exacerbation of asthma (acute asthma) and stable asthma.. We determined peripheral eosinophil count, and the serum levels of ECP and 27 types of cytokines/chemokines (IL-1 beta, IL-1 ra, IL-2, -4, -5, -6, -7, -8, -9, -10, -12, -13, -15 and -17, IFN-gamma, IP-10, TNF-alpha, GM-CSF, G-CSF, MCP-1, MIP-1 alpha and -1 beta, eotaxin, RANTES, PDGF-bb, FGF basic and VEGF) using a multiplex bead-based assay in 85 acute and 79 stable asthma patients, and 14 controls. We also examined the effects of systemic corticosteroids on these responses in acute asthma.. The serum levels of ECP, IL-5, -6, -8 and -10, G-CSF, MCP-1, IL-1 ra and IP-10 were significantly elevated in acute compared with stable asthma. Similarly, serum levels of ECP, IL-5 and IP-10 were significantly higher in acute asthma than in controls. Furthermore, in the acute phase, elevated serum levels of ECP, IL-5, IL-6, IL-1 ra and IP-10, but not IL-8, IL-10, G-CSF and MCP-1 were significantly reduced after treatments that included systemic corticosteroids.. Eosinophil activation could be induced by acute exacerbation of childhood asthma.

Topics: Adrenal Cortex Hormones; Asthma; Blood; Chemokine CCL2; Chemokine CXCL10; Chemokines; Child; Child, Preschool; Cytokines; Eosinophil Cationic Protein; Eosinophils; Female; Granulocyte Colony-Stimulating Factor; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-10; Interleukin-5; Interleukin-6; Interleukin-8; Leukocyte Count; Male

To elucidate the mechanisms of intractable pediatric bronchial asthma and the indication of low-dose erythromycin (EM) therapy, the serum chemokine levels of and the angiogenic factor were evaluated in 55 pediatric patients with bronchial asthma; 7.4 +/- 3.5 yr old, who had been treated with inhaled steroid, leukotriene receptor antagonist, theophylline and others for more than a year. Both the levels of interleukin (IL) 8 (p = 0.036) and vascular endothelial growth factor (VEGF) (p = 0.005) were higher in patients with severe type than those of patients with the milder type, while other chemokine levels such as serum eotaxin and MCP1 did not show the correlation with the severity of bronchial asthma. Induction of therapy with low-dose EM induced improvement of the clinical symptoms in patients with severe type and decrease of their serum chemokine levels: IL8; from 736 +/- 88 to 75 +/- 85 pg/ml (p < 0.0005), and VEGF; from 352.0 +/- 160.5 to 132.2 +/- 59.9 pg/ml (p = 0.021) within the next 6 months. Moreover, low-dose EM resulted in a decreased daily peak-trough fluctuation rate of the serum theophylline concentration; (C(max )- C(min))/C(min), from 1.3 +/- 0.5 to 0.5 +/- 0.3, which led to the maintenance of effective serum levels. These results indicated that IL8 and VEGF affect the severity of standard therapies resistance intractable bronchial asthma. Through the suppression of these chemokines and maintenance of effective theophylline levels, low-dose EM therapy improves the symptoms of bronchial asthma.

Topics: Anti-Asthmatic Agents; Anti-Bacterial Agents; Asthma; Chemokine CCL11; Child; Child, Preschool; Dose-Response Relationship, Drug; Erythromycin; Female; Humans; Interleukin-8; Leukotriene Antagonists; Male; Pediatrics; Theophylline; Treatment Outcome; Vascular Endothelial Growth Factor A

Human bronchial epithelial (HBE) cells contribute to asthmatic airway inflammation by secreting cytokines, chemokines, and growth factors, including interleukin (IL)-6, IL-8 and transforming growth factor (TGF) β1, all of which are elevated in asthmatic airways. This study examines the signaling pathways leading to TGFβ1 induced IL-6 and IL-8 in primary HBE cells from asthmatic and non-asthmatic volunteers. HBE cells were stimulated with TGFβ1 in the presence or absence of signaling inhibitors. IL-6 and IL-8 protein and mRNA were measured by ELISA and real-time PCR respectively, and cell signaling kinases by Western blot. TGFβ1 increased IL-6, but inhibited IL-8 production in both asthmatic and non-asthmatic cells; however, TGF induced significantly more IL-6 in asthmatic cells. Inhibition of JNK MAP kinase partially reduced TGFβ1 induced IL-6 in both cell groups. TGFβ1 induced Smad2 phosphorylation, and blockade of Smad2/3 prevented both the TGFβ1 modulated IL-6 increase and the decrease in IL-8 production in asthmatic and non-asthmatic cells. Inhibition of Smad2/3 also increased basal IL-8 release in asthmatic cells but not in non-asthmatic cells. Using CHIP assays we demonstrated that activated Smad2 bound to the IL-6, but not the IL-8 promoter region. We conclude that the Smad2/3 pathway is the predominant TGFβ1 signaling pathway in HBE cells, and this is altered in asthmatic bronchial epithelial cells. Understanding the mechanism of aberrant pro-inflammatory cytokine production in asthmatic airways will allow the development of alternative ways to control airway inflammation.

Topics: Adolescent; Adult; Aged; Asthma; Binding Sites; Blotting, Western; Bronchi; Case-Control Studies; Cells, Cultured; Chromatin Immunoprecipitation; Down-Regulation; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Female; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Male; Middle Aged; Phosphorylation; Polymerase Chain Reaction; Promoter Regions, Genetic; Protein Kinase Inhibitors; Respiratory Mucosa; RNA, Messenger; Signal Transduction; Smad2 Protein; Smad3 Protein; Transforming Growth Factor beta1; Up-Regulation; Young Adult

Thymic stromal lymphopoietin (TSLP) is an epithelial cell-derived cytokine that strongly activates dendritic cells and can initiate allergic inflammation. Since exposure to rhinovirus or double-stranded (ds) RNA (a surrogate of viral infection) induces TSLP expression in bronchial epithelial cells (BECs), this cytokine may link innate antiviral responses and the type 2 adaptive immune response.. As BECs from donors with asthma have a deficient interferon (IFN) response to rhinovirus infection, a study was undertaken to test the hypothesis that their antiviral response shows a bias towards TSLP production.. Primary BECs were grown from subjects with asthma and healthy volunteers. After exposure to dsRNA, interleukin (IL)-8, IFNbeta and TSLP mRNA and protein expression were measured by RT-qPCR and ELISA, respectively.. dsRNA dose-dependently increased IL-8 expression in BECs with no significant difference between the groups. However, BECs from subjects with asthma expressed less IFNbeta and more TSLP mRNA and protein in response to dsRNA than BECs from those without asthma (median (IQR) 57 (38-82) pg/ml vs 106 (57-214) pg/ml for IFNbeta (p<0.05) and 114 (86-143) pg/ml vs 65 (32-119) pg/ml for TSLP (p<0.05) in response to 10 microg/ml dsRNA for 24 h). Induction of TSLP mRNA by dsRNA was blocked by Toll-like receptor 3 or protein kinase inhibitors or by preventing de novo protein synthesis, but not by neutralisation of type I IFN receptors.. BECs from subjects with asthma are biased towards higher TSLP and lower IFNbeta production in response to dsRNA, suggesting that viral infection in asthma may lead to an altered mediator profile that biases towards a Th2 immune response.

Topics: Adult; Asthma; Bronchi; Bronchoscopy; Cells, Cultured; Cytokines; Dose-Response Relationship, Immunologic; Epithelial Cells; Female; Gene Expression Regulation; Humans; Immunity, Innate; Interferon-beta; Interleukin-8; Male; Middle Aged; Protein Kinase Inhibitors; Respiratory Mucosa; RNA, Double-Stranded; RNA, Messenger; Thymic Stromal Lymphopoietin; Toll-Like Receptor 3; Young Adult

Neutrophils are potent contributors to the lung pathophysiological changes occurring in allergic airway inflammation, which typically involve T helper type 2 (Th2) cytokine overexpression. We have previously reported that equine pulmonary endothelial cells are activated by the Th2 cytokine IL-4 and express chemotactic factors for neutrophils after stimulation. We have further explored the possible mechanisms linking Th2-driven inflammation and neutrophilia by studying the effects of recombinant equine IL-4, a prototypical Th2 cytokine, on peripheral blood neutrophils (PBN) isolated from normal animals and from horses with asthmatic airway inflammation (equine heaves). We found that IL-4 induced morphological changes in PBN, dose- and time-dependent expression of IL-8 mRNA, as well as the release of chemotactic factors for neutrophils in culture supernatants. Also, IL-4 induced a mixed inflammatory response in PBN from control and asthmatic-animals with increased expression of proinflammatory IL-8 and TNF-α but a marked inhibition of IL-1β. IL-4 type I receptor (IL-4Rα) and CD23 (FcεRII) expression were also upregulated by IL-4. Importantly, disease as well as chronic antigenic exposure modified gene expression by PBN. Finally, we found that activation of equine neutrophils with IL-4 involved STAT6 phosphorylation and p38 MAPK and phosphatidylinositol 3-kinase (PI3K); the pharmacological inhibitors, SB-203580 and LY-294002, respectively, significantly reversed IL-4-induced gene modulation in PBN. Overall, results from this study add to the link between Th2-driven inflammation and neutrophilia in the equine model and further extend the characterization of IL-4 effects on neutrophils.

Topics: Animals; Asthma; Blotting, Western; Chemotaxis, Leukocyte; Gene Expression Profiling; Horses; Immunoenzyme Techniques; Inflammation; Interleukin-4; Interleukin-8; Neutrophil Activation; p38 Mitogen-Activated Protein Kinases; Phosphatidylinositol 3-Kinases; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; STAT6 Transcription Factor; Tumor Necrosis Factor-alpha

The association between vitamin D deficiency and asthma epidemic has been recognized. Tumor necrosis factor (TNF)-alpha and chemokines play important roles in pathogenesis of asthma. However, whether vitamin D has immunoregulatory function on TNF-alpha and chemokines expression in human monocytes is still unknown. The human monocytic cell line, THP-1 cells and human primary monocytes were pretreated with various concentration of 1alpha,25-(OH)(2)D(3) for 2 h before stimulation with lipopolysaccharide (LPS). Supernatants were collected 24 or 48 h after LPS stimulation. The levels of TNF-alpha, interferon-inducible protein 10 (IP-10)/CXCL10 (the Th1-related chemokine), macrophage-derived chemokine (MDC)/ CCL22 (the Th2-related chemokine), and interleukin 8 (IL-8)/CXCL8 (the neutrophil chemoattractant) were measured by ELISA. 1alpha,25-(OH)(2)D(3) could significantly suppress TNF-alpha and IP-10 expression in LPS-stimulated THP-1 and human primary monocytes. However, 1alpha,25-(OH)(2)D(3), especially in higher concentration, could significantly enhance MDC expression. 1alpha,25-(OH)(2)D(3) had no significant effects on IL-8 expression. We found 1alpha,25-(OH)(2)D(3) could significantly suppress TNF-alpha and Th1-related chemokine IP-10, which both play important roles in pathogenesis of severe refractory asthma and autoimmune diseases. However, 1alpha,25-(OH)(2)D(3) enhanced Th2-related chemokine MDC, which may result in Th2 inflammatory cell recruitment and thus adversely affect asthmatic patients. Although vitamin D has potential utility in the treatment of asthma and autoimmune diseases, excessive use of vitamin D may not be suitable in patients with Th2 allergic diseases.

Topics: Asthma; Autoimmune Diseases; Calcitriol; Cell Line; Cells, Cultured; Chemokine CCL22; Chemokine CXCL10; Chemokines; Cholecalciferol; Down-Regulation; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-8; Lipopolysaccharides; Monocytes; Osmolar Concentration; Time Factors; Tumor Necrosis Factor-alpha; Up-Regulation; Vitamin D Deficiency

The mechanism of action involved in how Dermatophagoides pteronyssinus (Der p) 1 initiates the nasal allergic cascade is poorly understood.. We detected proinflammatory cytokine production (GM-CSF, TNF-α, IL-1β, IL-6, and IL-8) and associated signal molecules in primarily cultured nasal epithelial cells (NECs) from patients with allergic rhinitis (AR) after Der p1 stimulation, using ELISA, RT-PCR, and Western blot. We also evaluated the importance of protease-activated receptors (PAR)/phosphatidylinositol 3 kinase (PI3K)/NFκB signaling pathways in IL-6 and IL-8 production using glucocorticoids and specific inhibitors, LY294002 and PDTC.. We observed significantly elevated IL-6 and IL-8 production (both gene and protein) in NECs after Der p1 stimulation, and demonstrated that the expressions of PAR2, pAkt, and pp65 were upregulated after Der p1 stimulation, which were associated with IL-6 and IL-8 production in NECs.. These findings demonstrate that the PAR/PI3K/NFκB signaling pathway is involved in the induction of IL-6 and IL-8 in Der-p1-stimulated NECs from AR patients, and may have potential implications for the prevention and treatment of AR and asthma.

Topics: Adult; Antigens, Dermatophagoides; Arthropod Proteins; Asthma; Cells, Cultured; Cysteine Endopeptidases; Female; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Nasal Mucosa; NF-kappa B; Phosphatidylinositol 3-Kinases; Receptor, PAR-1; Receptor, PAR-2; Rhinitis, Allergic, Perennial; RNA, Messenger; Signal Transduction; Young Adult

Cockroaches are potent aeroallergens associated with asthma. Several reports suggest that a novel group of G protein-linked receptors, protease-activated receptors (PARs), may be involved in the intracellular signaling pathway induced by aeroallergens of the epithelial cells.. To investigate the mechanisms of American cockroach allergens (CraA) on interleukin 8 (IL-8) in human pulmonary epithelial cells.. Protease activities of CraA were quantified by the Azocoll method. The gene and protein expressions of IL-8 from CraA-stimulated A549 cells were quantified by real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The activity of different mitogen-activated protein kinases (MAPKs) was assessed by Western blot.. CraA-induced A549 cell IL-8 secretion in a dose-dependent manner at both the messenger RNA and protein levels. CraA-induced IL-8 secretion can be blocked by serine protease inhibitors, phenylmethane sulfonyl fluoride, and aprotinin but not by other protease inhibitors. Blocking antibodies against the cleavage sites of PAR-2 and PAR-3, but not of PAR-1, inhibited CraA-induced IL-8 production. CraA induced significant PAR-2 and PAR-3 messenger RNA upregulation and extracellular-regulated kinase (ERK/1/2) and Jun N-terminal kinase (JNK) phosphorylation but not p38 MAPK. Furthermore, ERK1/2 (U0126) and JNK (SP600125) inhibitors inhibited CraA-induced IL-8 secretion by 100% and 45%, respectively.. Both PAR-2 and PAR-3 might play a role in CraA-induced IL-8 secretion from human airway epithelial cells. It signals mainly through the ERK1/2 and partly from the JNK pathways. The key receptors and signaling molecules mediate cytokine release from the respiratory epithelium and can be potential therapeutic targets in treating cockroach allergy.

Topics: Adaptor Proteins, Signal Transducing; Allergens; Animals; Antigens, Plant; Asthma; Cell Cycle Proteins; Enzyme Inhibitors; Extracellular Signal-Regulated MAP Kinases; Humans; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Membrane Proteins; p38 Mitogen-Activated Protein Kinases; Periplaneta; Receptor, PAR-1; Respiratory Mucosa; Signal Transduction; Up-Regulation

despite a number of important differences in the pathogenesis, course, and prognosis, asthma and chronic obstructive pulmonary disease (COPD) have many features in common. Furthermore, smoking induces considerable overlap in pathogenesis and clinical features between these conditions. This study aimed to reveal what inflammatory patterns prevail in clinically established diagnosis groups, including overlap phenotypes of asthma and COPD, and to evaluate the correlation with airway reversibility and hyperreactivity in these overlapping conditions.. a total of 110 patients (17 healthy subjects; 16 "healthy" smokers; 46 asthma patients: 24 smokers and 22 non-smokers; and 31 COPD patients: 10 COPD patients with reversibility and 21 without) participated in the study. Induced sputum, reversibility testing, methacholine and adenosine 5'monophosphate (AMP) provocation challenges, and skin prick testing were performed. Airways inflammation was assessed by differential cell counts, and cytokines (interleukin-8 [IL-8] and tumor necrosis factor-alpha [TNF-α]) were measured in induced sputum by enzyme-linked immunosorbent assay (ELISA).. COPD patients with reversibility had increased sputum neutrophils, IL-8, and TNF-α levels compared to smoking asthmatics. No difference was found in inflammatory cells and cytokines between COPD subgroups. Sputum neutrophilia was inversely correlated with the change in forced expiratory volume in one second (ΔFEV(1)) in smoking asthmatic patients (r = -0.563, P = 0.036). No correlation was found between airway hyperresponsiveness (AHR), either with methacholine or AMP, and inflammation in asthmatic patients, regardless of smoking. Reversibility was not correlated with inflammation in COPD patients. However, the response to AMP challenge was correlated with sputum neutrophils (r = 0.844, P = 0.001).. although overlaps exist in the disease characteristics of asthma and COPD, the combination of lung function testing, sputum induction, and AHR reveals information that facilitates the distinction between these diseases, allowing clinicians to better tailor their therapy.

Topics: Adult; Asthma; Bronchial Hyperreactivity; Cytokines; Diagnosis, Differential; Diagnostic Tests, Routine; Humans; Interleukin-8; Middle Aged; Phenotype; Pulmonary Disease, Chronic Obstructive; Respiratory Function Tests; Smoking; Spirometry; Sputum; Tumor Necrosis Factor-alpha

IL-33 is a member of the IL-1 family and mediates its biological effects via the ST2 receptor, which is selectively expressed on Th2 cells and mast cells. Although polymorphic variation in ST2 is strongly associated with asthma, it is currently unclear whether IL-33 acts directly on lung tissue cells at sites of airway remodeling. Therefore, we aimed to identify the IL-33-responsive cells among primary human lung tissue cells. ST2 mRNA was expressed in both endothelial and epithelial cells but not in fibroblasts or smooth muscle cells. Correspondingly, IL-33 promoted IL-8 production by both endothelial and epithelial cells but not by fibroblasts or smooth muscle cells. Transfection of ST2 small interference RNA into both endothelial and epithelial cells significantly reduced the IL-33-dependent upregulation of IL-8, suggesting that IL-33-mediated responses in these cells occur via the ST2 receptor. Importantly, Th2 cytokines, such as IL-4, further enhanced ST2 expression and function in both endothelial and epithelial cells. The IL-33-mediated production of IL-8 by epithelial cells was almost completely suppressed by corticosteroid treatment. In contrast, the effect of corticosteroid treatment on the IL-33-mediated responses of endothelial cells was only partial. IL-33 induced activation of both ERK and p38 MAPK in endothelial cells but only ERK in epithelial cells. p38 MAPK was required for the IL-33-mediated responses of endothelial cells, whereas ERK was required for IL-33-mediated IL-8 production by epithelial cells. Taken together, these findings suggest that IL-33-mediated inflammatory responses of lung tissue cells may be involved in the chronic allergic inflammation of the asthmatic airway.

Topics: Asthma; Blotting, Western; Cells, Cultured; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Fibroblasts; Humans; Inflammation; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Interleukin-8; Interleukins; Lung; Myocytes, Smooth Muscle; Receptors, Cell Surface; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering; Signal Transduction; Transfection

Neutrophilic asthma is thought to be less responsive than eosinophilic asthma to anti-inflammatory therapies including corticosteroids. Chlamydia pneumoniae has been implicated in asthma, possibly by induction of interleukin (IL-8). We hypothesized that IL-8 is increased in the bronchoalveolar lavage (BAL) fluid from children with asthma and C. pneumoniae.. BAL fluid was analyzed for C. pneumoniae and IL-8 using polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay from 2 asthma patient populations in the Bronx, NY and Massachusetts with an average age of 8 and 8.7 years old, respectively. For comparison, samples were also analyzed for C. trachomatis and Mycoplasma 16s DNA.. Of 18 Bronx samples analyzed, 6 (33%) were PCR-positive for C. pneumoniae, 10 (56%) for C. trachomatis, and 8 (44%) for Mycoplasma 16s DNA. IL-8 from C. pneumoniae-positive samples was 3.3-fold higher compared with negative samples (P = 0.003). There was no difference between patients tested for C. trachomatis or Mycoplasma. Of 84 Massachusetts samples analyzed, 42 (50%) were PCR-positive for C. pneumoniae, 42 (50%) for C. trachomatis, and 13 (16%) for Mycoplasma. IL-8 concentration from C. pneumoniae-positive samples was 10.49-fold higher compared with negative samples (P = 0.0001). As in the Bronx cohort, there were no differences between patients tested for C. trachomatis or Mycoplasma. Lastly, BAL neutrophilia predicted the presence of C. pneumoniae but not Mycoplasma or C. trachomatis.. Children with asthma who were PCR-positive for C. pneumoniae demonstrated elevated concentrations of IL-8 and neutrophils in BAL fluid compared with similar patients who were positive for C. trachomatis or Mycoplasma organisms, but PCR-negative for C. pneumoniae. Undiagnosed C. pneumoniae infection in children may therefore contribute to poorly controlled asthma via induction of IL-8.

Topics: Asthma; Bronchoalveolar Lavage Fluid; Child; Chlamydia trachomatis; Chlamydophila Infections; Chlamydophila pneumoniae; DNA, Bacterial; DNA, Ribosomal; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Male; Massachusetts; Mycoplasma; Neutrophils; New York City; Polymerase Chain Reaction; Respiratory System; RNA, Ribosomal, 16S

Chronic obstructive pulmonary disease (COPD) is a heterogeneous disease characterized by a combination of 3 different disorders, namely chronic asthma, chronic bronchitis and pulmonary emphysema, sometimes simultaneously present in the same subject.. The aim of our study was to compare sputum inflammatory markers in patients with different phenotypes of chronic airway obstruction.. Forty-five subjects (forced expiratory volume in 1 s/vital capacity, FEV(1)/VC: 58.8 +/- 12.2%; FEV(1): 49.8 +/- 11.5% of predicted) were classified as chronic asthma (n = 10) or COPD patients (n = 35); the latter were further divided into patients with prevalent chronic bronchitis (n = 24) or prevalent pulmonary emphysema (n = 11) according to clinical history and functional evaluation, and underwent sputum induction and analysis of inflammatory cell and soluble mediators.. Patients with chronic asthma showed higher sputum eosinophil percentages and eosinophilic cationic protein levels, and lower neutrophil percentages and neutrophil elastase levels than COPD patients. Neutrophil chemotactic activity in sputum supernatant was higher than the pool of normal subjects both in chronic asthma and COPD patients. No difference in sputum cell composition and levels of soluble mediators was observed between patients with chronic bronchitis and patients with pulmonary emphysema.. The pattern of airway inflammation in induced sputum of patients with chronic asthma is different from that of COPD patients with a similar FEV(1). Among COPD patients, however, the pattern of airway inflammation shows no difference between chronic bronchitis and patients with pulmonary emphysema, suggesting that these two clinically and functionally distinct phenotypes share a common inflammatory pattern as detected by induced sputum.

Topics: Aged; Asthma; Biomarkers; Bronchitis, Chronic; Eosinophil Cationic Protein; Female; Granulocytes; Humans; Interleukin-8; Leukocyte Elastase; Male; Middle Aged; Phenotype; Pulmonary Emphysema; Sputum

Individuals with asthma are prone to viral and bacterial infections, and most asthma exacerbations have been linked to viruses, particularly rhinovirus. Excess transforming growth factor (TGF)-beta present in asthmatic airways may cause immune suppression, as well as transdifferentiate fibroblasts to myofibroblasts, thereby augmenting proinflammatory responses after rhinovirus infection. After rhinovirus infection we examined virus replication and host cell immune responses in airway fibroblasts in the presence of TGF-beta1 and in myofibroblasts. Primary culture fibroblasts were pretreated with TGF-beta1 or transdifferentiated into myofibroblasts, and then infected with rhinovirus. Viral replication, virus release, chemokine production, and interferon (IFN) responses were measured over 72 hours. Rhinovirus replication and virus release into supernatants were enhanced in fibroblasts incubated with TGF-beta1 and in fibroblasts obtained from patients with asthma. Myofibroblasts also showed more rhinovirus replication, and infected myofibroblasts produced excess neutrophil chemokines. Examination of innate responses revealed blunting of type I IFN reactions with dissociated viral RNA and IFN mRNA responses. Addition of type I IFN restituted antiviral responses, and the effect of TGF-beta1 appeared to be mediated via actions on IFN regulatory factor-3 pathways. These data demonstrate that TGF-beta1 mediates enhanced virus replication and proinflammatory responses in airway cells. TGF-beta may act as an endogenous immunosuppressant promoting virus replication and inflammation during the evolution of acute severe asthma associated with rhinovirus infection.

Topics: Adult; Asthma; Cell Death; Cell Differentiation; Cells, Cultured; Chemokines; Child; Fibroblasts; Humans; Immunity, Innate; Interferon Regulatory Factor-3; Interferon Type I; Interleukin-8; Neutrophils; Picornaviridae Infections; Respiratory Mucosa; Rhinovirus; Signal Transduction; Transforming Growth Factor beta1; Virus Replication

Airway angiogenesis may be an important part of structural remodelling in the pathogenesis of asthma. The development of asthma is frequently preceded by rhinitis.. We sought to determine whether the levels of angiogenesis-related factors are elevated in airways of patients with rhinitis or controlled asthma.. We analysed the induced sputum of 18 rhinitis patients, 16 asthmatic patients, and 15 healthy controls. The concentrations of angiogenin, vascular endothelial growth factor (VEGF), IL-8, fibroblast growth factor (bFGF), and TNF-alpha were measured by cytometric bead arrays.. We found significantly increased angiogenin and VEGF concentrations in the induced sputum supernatant of both rhinitis and asthma patients compared with that of the healthy control group (P< or =0.0005). With the exception of TNF-alpha, there was no difference in the other angiogenic factors; TNF-alpha levels were higher in the rhinitis group than in the control group (P=0.02).. These in vivo results suggest increased airway angiogenesis in patients with rhinitis without asthma as well as in corticosteroid-treated and well-controlled asthma patients.

Topics: Adolescent; Adult; Aged; Angiogenesis Inducing Agents; Asthma; Cell Count; Eosinophils; Female; Fibroblast Growth Factor 2; Humans; Interleukin-8; Lymphocytes; Macrophages; Male; Middle Aged; Neutrophils; Rhinitis; Ribonuclease, Pancreatic; Sputum; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Cigarette smoking in asthma patients causes insensitivity to inhaled glucocorticoids (GCs). We tested the hypothesis that smoking causes GC insensitivity in alveolar macrophages (AMs) obtained from patients with asthma.. Nineteen asthmatic nonsmokers (ANSs) and 13 asthmatic smokers (ASMs) underwent BAL. AMs were cultured with or without dexamethasone, 0.1 to 1,000 nmol/L, for 2 h before lipopolysaccharide (LPS) [1 microg/mL] stimulation. After 6 h, supernatants were harvested for enzyme-linked immunosorbent assay, and messenger RNA was collected for real-time (RT)-polymerase chain reaction (PCR).. ASMs had higher numbers of AMs per milliliter of BAL fluid than ANSs (1.98 vs 0.75 x 10(6) cells/mL, respectively; p = 0.007). Cigarette smoking significantly attenuated the LPS response for all three cytokines tested among ANSs vs ASMs (tumor necrosis factor [TNF]-alpha, 31.6 vs 10.6 ng/mL, respectively (p = 0.01); interleukin [IL]-6, 25.8 vs 10.8 ng/mL, respectively (p = 0.002); IL-8, 62.5 vs 36.1 ng/mL, respectively (p = 0.001)). There was no difference in dexamethasone dose-response curves between ANSs and ASMs (p > 0.05 for all comparisons). The inhibitory concentration of 50% (IC(50)) for IL-6 was 120.6 vs 83.3, respectively, and for TNF-alpha it was 4.9 vs 8.6, respectively; an IC(50) was not achieved for IL-8. RT-PCR also showed no difference in the suppression of cytokine messenger RNA levels between groups, with IL-8 being the most GC-insensitive cytokine.. Cigarette smoking in patients with asthma increases the number of airway AMs and attenuates their response to LPS, which may have implications in host immune function. Cigarette smoking does not alter the GC sensitivity of AMs in patients with asthma. There was differential cytokine sensitivity, with IL-8 being the least GC-sensitive cytokine. GC-insensitive IL-8 production from AMs may be a mechanism by which neutrophils are attracted into the airways.

Topics: Adult; Asthma; Case-Control Studies; Cell Culture Techniques; Dexamethasone; Female; Glucocorticoids; Humans; Interleukin-6; Interleukin-8; Lipopolysaccharides; Macrophages, Alveolar; Male; Middle Aged; Smoking; Tumor Necrosis Factor-alpha; Young Adult

Human rhinoviruses (HRV) are a major cause of asthma exacerbations and hospitalization. Studies using primary cultures suggest that this may be due to impaired production of type I and type III IFNs by asthmatic bronchial epithelial cells. Although epithelial cells are the main target for HRV infection, HRV can be detected in the subepithelial layer of bronchial mucosa from infected subjects by in situ hybridization. Therefore, we postulated that submucosal fibroblasts are also involved in the innate antiviral response to HRV infection in asthma. We found that regardless of subject group, bronchial fibroblasts were highly susceptible to RV1b infection. IL-8 and IL-6 were rapidly induced by either HRV or UV-irradiated virus, suggesting that these responses did not require viral replication. In contrast, RANTES expression was dependent on viral replication. Regardless of disease status, fibroblasts did not respond to HRV infection with significant induction of IFN-beta, even though both groups responded to synthetic dsRNA with similar levels of IFN-beta expression. Exogenous IFN-beta was highly protective against viral replication. Our data suggest that fibroblasts respond to HRV with a vigorous proinflammatory response but minimal IFN-beta expression. Their susceptibility to infection may cause them to be a reservoir for HRV replication in the lower airways, especially in asthmatic subjects where there is reduced protection offered by epithelial-derived IFNs. Their ability to support viral replication coupled with their vigorous proinflammatory response following infection may contribute to asthma exacerbations.

Topics: Adolescent; Adult; Asthma; Bronchi; Cells, Cultured; Chemokine CCL5; Cytopathogenic Effect, Viral; Female; Fibroblasts; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Male; Middle Aged; Picornaviridae Infections; Rhinovirus; Tumor Necrosis Factor-alpha; Young Adult

Several chronic respiratory diseases exhibit hyperactive immune responses in the lung: abundant inflammatory mediators; infiltrating neutrophils, macrophages, lymphocytes and other immune cells; and increased level of proteases. Such diseases include cystic fibrosis (CF), chronic obstructive pulmonary disease (COPD) and severe/neutrophilic asthma. Paradoxically, patients with these diseases are also susceptible to detrimental bacterial infection and colonization. In this paper, we seek to explain how a positive feedback mechanism via IL-8 could lead to desensitization of epithelial cells to pathogen recognition thus perpetuating bacterial colonization and chronic disease states in the lung. Such insight was obtained from mathematical modeling of the IRAK/TRAF6 signaling module, and is consistent with existing clinical evidence. The potential implications for targeted treatment regimes for these persistent respiratory diseases are explored.

Topics: Asthma; Epithelial Cells; Humans; Interleukin-1 Receptor-Associated Kinases; Interleukin-8; Lung; Models, Biological; Pulmonary Disease, Chronic Obstructive; Respiration Disorders; Signal Transduction; TNF Receptor-Associated Factor 6

House dust mites (HDM) are well-known as a source of indoor aeroallergens and for causing allergic airway diseases. Some proteolytic HDM allergens are known to activate respiratory epithelial cells to produce pro-inflammatory mediators, while there is limited knowledge regarding such activity among non-proteolytic HDM allergens.. To investigate whether Der p 2, a major non-proteolytic allergen of Dermatophagoides pteronyssinus, activates respiratory epithelial cells to produce mediators involved in asthma pathogenesis and to elucidate the mechanism of such activation.. The human bronchial epithelial cell line BEAS-2B, normal human bronchial epithelial (NHBE) cells and the alveolar epithelial cell line A549 were exposed to recombinant Der p 2. Following exposure, we analysed a panel of soluble mediators and cell adhesion receptors involved in asthma pathogenesis by promoting recruitment, survival and binding of inflammatory cells. The involvement of nuclear factor (NF)-kappaB and mitogen-activated protein kinases (MAPKs) was studied using specific inhibitors.. Der p 2 activated bronchial BEAS-2B and NHBE cells, but not alveolar A549 cells. In BEAS-2B cells Der p 2 induced dose-dependent up-regulation in both mRNA level and protein secretion of granulocyte-macrophage colony-stimulating factor, IL-6, IL-8, monocyte-chemotactic protein-1 and macrophage inflammatory protein-3alpha. Secretion as well as surface expression of intercellular adhesion molecule (ICAM)-1 was also up-regulated, which was associated with increased adhesion of monocytes to the epithelial cells. The release of cytokines and chemokines was regulated by NF-kappaB and MAPK activation in different ways, while expression of ICAM-1 was solely dependent on NF-kappaB activation.. These results show that Der p 2 activates respiratory epithelial cells, indicating that this non-proteolytic allergen, in addition to its immunogenic properties, can aggravate respiratory airway disease by adjuvant-like activation of the lung epithelium.

Topics: Animals; Antigens, Dermatophagoides; Arthropod Proteins; Asthma; Bronchi; Cells, Cultured; Chemokine CCL2; Chemokine CCL20; Dermatophagoides pteronyssinus; Epithelial Cells; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; Mitogen-Activated Protein Kinases; NF-kappa B; RNA, Messenger; Signal Transduction; Up-Regulation

There is evidence that eosinophils and neutrophils are simultaneously increased in the airways of some patients with chronic refractory asthma. The mechanisms by which neutrophils accumulate in the airways of asthmatics remain to be elucidated, however, chemoattractants for neutrophils such as CXC chemokines may affect either the accumulation or functional status of neutrophils in such patients. The objective of the present study was to identify the CXC chemokine responsible for the neutrophilic and possibly eosinophilic inflammation observed in the airways of patients with refractory asthma.. Following the inhalation of hypertonic saline, induced sputum was obtained from 14 healthy controls, 16 patients with mild well-controlled nonrefractory asthma, and 14 patients with refractory asthma. Concentrations of CXC chemokines and differential inflammatory cell counts were determined.. The percentages of induced sputum eosinophils were significantly higher both in patients with nonrefractory asthma and in patients with refractory asthma. On the other hand, the percentages of neutrophils were increased only in sputum from patients with refractory asthma. The concentration of IL-8, but not ENA-78 or GRO-alpha, was also significantly increased in induced sputum from patients with refractory asthma. The concentration of IL-8 correlated significantly with the percentages of neutrophils.. The results of the present study suggest that IL-8, but not ENA-78 or GRO-alpha, may contribute to the observation of neutrophilic inflammation in patients with refractory asthma.

Topics: Asthma; Eosinophils; Female; Humans; Inflammation; Interleukin-8; Leukocyte Count; Male; Middle Aged; Neutrophils; Sputum

CXCL8 is a neutrophil and mast cell chemoattractant that is involved in regulating inflammatory cell influx in asthma. Here, we investigated the transcriptional mechanism involved in CXCL8 induction by TNF-alpha in cultured human airway smooth muscle (HASM) cells and compared these in cells from nonasthmatic and asthmatic individuals. Transfection studies with mutated CXCL8 promoter constructs identified NF-kappaB, activating protein-1, and CAAT/enhancer binding protein (C/EBP)beta as key transcription factors, and binding of these three transcription factors to the CXCL8 promoter after TNF-alpha stimulation was confirmed by chromatin immunoprecipitation analysis. Cells derived from asthmatic individuals produced significantly higher levels of CXCL8 than nonasthmatic cells both basally and following 24 h of stimulation with TNF-alpha (p < 0.001). Furthermore, chromatin immunoprecipitation studies detected increased binding of NF-kappaB p65 and RNA polymerase II to the CXCL8 promoter of asthmatic HASM cells both in the presence and absence of TNF-alpha stimulation. This was not due to either an increased activation or phosphorylation of NF-kappaB per se or to an increase in its translocation to the nucleus. Increased binding of C/EBPbeta to the CXCL8 promoter of unstimulated cells was also detected in the asthmatic HASM cells. Collectively these studies show that HASM cells from asthmatic individuals have increased CXCL8 production due to the presence of a transcription complex on the CXCL8 promoter, which contains NF-kappaB, C/EBPbeta, and RNA polymerase II. This is the first description of an abnormality in transcription factor binding altering chemokine expression in airway structural cells in asthma.

Topics: Asthma; Bronchi; CCAAT-Enhancer-Binding Protein-beta; Cells, Cultured; Female; Humans; Interleukin-8; Male; Middle Aged; Myocytes, Smooth Muscle; Protein Binding; RNA Polymerase II; Transcription Factor AP-1; Transcription Factor RelA; Transcription, Genetic; Tumor Necrosis Factor-alpha; Up-Regulation

Topics: Adult; Asthma; Biomarkers; Female; Humans; Inflammation; Inflammation Mediators; Interleukin-13; Interleukin-8; Leukocytes; Male; Respiratory Mucosa; Severity of Illness Index; Sputum

To evaluate the relationship between pro-inflammatory and pro-remodeling mediators and severity and control of asthma in children, the levels of IL-8, MMP-9, TIMP-1 in induced sputum supernatants, the number of sputum eosinophils, as well as FeNO, were investigated in 35 asthmatic children, 12 with intermittent (IA) and 23 with moderate asthma (MA), and 9 controls (C). The patients with asthma were followed for 1 yr and sputum was obtained twice during the follow-up. Biomarker levels were correlated with the number of exacerbations. We found that IL-8, MMP-9, TIMP-1 and the numbers of eosinophils in induced sputum, as well as FeNO, were increased in children with IA and MA in comparison to C. The ongoing inflammation was confirmed by increased nuclear p65 NF-kappaB subunit localization in sputum cells. In MA, FeNO measurements, sputum eosinophils and IL-8 levels, positively correlated with the occurrence of disease exacerbations during a 1-yr follow-up. According to FeNO, sputum eosinophils and IL-8 sputum concentrations, and the number of exacerbations, two distinct phenotypes of MA were identified. This study shows that the presence of bronchial inflammation is detectable in the airways of some IA, as well as in the airways of MA, despite the regular ICS treatment. This study also proposes the need to perform large prospective studies to confirm the importance of measuring specific biomarkers in induced sputum, concomitantly to FeNO analyses, to assess sub-clinical airway inflammation and disease control in children with asthma.

Topics: Adolescent; Asthma; Biomarkers; Bronchitis; Child; Disease Progression; Eosinophils; Female; Follow-Up Studies; Humans; Interleukin-8; Leukocyte Count; Male; Matrix Metalloproteinase 9; Sputum; Tissue Inhibitor of Metalloproteinase-1

To observe the airway inflammatory change in asthmatic rats complicated with chronic obstructive pulmonary disease (COPD) and to assess the intervention effects of Chinese herbs for reinforcing Shen and supplementing qi (CH) on it.. Eighty-four Norway rats were randomized into 7 groups, the normal control group (A), the COPD model group (B), the asthma model group (C), the combined COPD and the asthma model group (D), and the three CH treated groups (E, F and G, combined model rats administered by low-, moderate- and high- dose CH, respectively), 12 rats in each group. Changes of symptoms, pathologic changes of the lung tissue, airway reactivity, and serum levels of interleukin-4, interleukin-6, interleukin-8 (IL-4, IL-6, and IL-8) and interferon-gamma (IFN-gamma) in rats were observed.. Symptoms were alleviated in the three CH treated groups. Similar pathological features were shown in group B and D, showing inflammatory cell, mainly lymphocyte, infiltration in bronchial and lung tissues, with cilia denudation, partial alveolar wall rupture, alveolar cavity expansion, and accompanied with evident eosinophilic infiltration. These inflammatory exudation in group E-G was alleviated, while in group C, it developed showing a trend similar to that in group D. Airway resistance raised along with the concentration of Mch used. In group D, the serum level of IL-4 was higher than that in group B, and level of INF-gamma was lower than that in group A, B and C (all P <0.05). CH showed a lowering effect on serum levels of IL-4 and -8, and a dose-dependent rising effect on IFN-gamma.. IL- 4 significantly increased and INF-gamma decreased in rat model of combined COPD and asthma, its mechanism is similar to that of Th1/Th2 imbalance in asthma. Chinese herbs for reinforcing Shen and supplementing qi could improve the symptoms and inhibit the airway inflammation in the combined COPD and asthma model rats, its mechanism might be related with the alleviation of TH1/TH2 imbalance.

Topics: Animals; Asthma; Drugs, Chinese Herbal; Inflammation; Interferon-gamma; Interleukin-4; Interleukin-6; Interleukin-8; Male; Phytotherapy; Pulmonary Disease, Chronic Obstructive; Qi; Rats; Rats, Inbred BN

Acute irritant-induced asthma (IrIa) or reactive airways dysfunction syndrome is caused by exposure to a high concentration of an agent. The long-term pathologic consequences of IrIa remain thus far unknown.. The aim of our study was to investigate the chronic airway inflammation and remodeling that occur in association with IrIa.. Ten subjects with a history of IrIa (mean interval of 10.9 years, minimum of 4 years, since the inhalational accident) underwent bronchoscopy followed by bronchoalveolar lavage and bronchial biopsies. Immunologic and morphologic data from patients with IrIa were compared with those of patients with mild to moderate asthma as well as healthy controls.. Bronchoalveolar lavage fluid analysis showed increased eosinophil and neutrophil counts in 30% and 60% of subjects with IrIa, respectively. In the supernatant of bronchoalveolar lavage, we found a significant increase in the majority of mediators compared with healthy subjects and a significant increase in eosinophilic cationic protein, IL-8, basic fibroblast growth factor, and matrix metalloproteinase 1 compared with control patients with asthma. Evaluation of basement membrane thickness (subepithelial fibrosis) demonstrated a significant increase in patients with IrIa compared with healthy subjects and subjects with asthma. Basement membrane thickness also significantly correlated with the PC(20) value. The epithelial cell detachment showed an elevated although not significant trend compared with subjects with asthma and control subjects. Immunocytochemical analysis demonstrated increases in the number of eosinophil cationic protein and TGF-beta1-positive cells compared with healthy controls.. This study provides evidence of a significant eosinophilic and neutrophilic inflammation as well as remodeling in IrIa many years after an inhalational accident.

Topics: Acute Disease; Adult; Aged; Asthma; Bronchoalveolar Lavage Fluid; Chronic Disease; Eosinophil Cationic Protein; Eosinophils; Female; Fibroblast Growth Factors; Humans; Inflammation; Interleukin-8; Irritants; Lung; Male; Matrix Metalloproteinase 1; Middle Aged; Neutrophils; Surveys and Questionnaires; Transforming Growth Factor beta1

To investigate the changes of neutrophils in airway inflammation in children with severe asthma.. Children with mild to moderate asthma (n=23), severe asthma (n=16) and healthy control subjects (n=16) underwent lung function tests and sputum induction. The sputum specimens were assayed for cellular differential count, the supernatant and peripheral blood were assayed for the concentrations of IL-8 by "sandwich" enzyme linked immunosorbent assay (ELISA). Sputum supernatant, IL-8 and mifepristone were assessed for their abilities to prolong the in vitro survival of blood-derived neutrophils.. The percentage of sputum neutrophils was significantly higher in severe asthmatics [59.54 (41.99-74.65)%] than mild-moderate asthmatics [30.03 (15.94-47.71)%] and healthy control subjects [29.72(16.53-45.74)%] (P < 0. 01); the level of IL-8 in sputum was significantly higher in severe asthmatics [2907.78 (331.67 - 3457.93) ng/L] than mild-moderate asthmatics [287.58 (130.75-656.84) ng/L] and healthy control subjects [179.2 (58.55-346.59) ng/L] (P < 0.01); the percentages of neutrophilic apoptosis respectively cultured with LPS [(10.57 +/- 1.97)%], severe asthmatics supernatant [(11.82 +/- 2.96)%], IL-8 [(10.47 +/- 1.93)%], dexamethasone [(9.93 +/- 1.95)%], severe asthma supernatant + mifepristone [(12.15 +/- 2.86)%] in vitro were lower than that cultured with PBS [(17.98 +/- 2.27)%], healthy control supernatant [(17.37 +/- 2.50)%], mild-moderate asthmatics supernatant [(16.35 +/- 3.26)%], mifepristone [(17.89 +/- 2.38)%], and dexamethasone + mifepristone [(17.06 +/- 2.59)%] (P < 0.01).. Suppression of neutrophilic apoptosis seems to play a potential role in airway neutrophilic inflammation in severe asthmatics, and the level of IL-8 in sputum was significantly higher in patients with severe asthmatics.

Topics: Adolescent; Apoptosis; Asthma; Case-Control Studies; Child; Female; Humans; Inflammation; Interleukin-8; Leukocyte Count; Male; Neutrophils; Respiratory System; Sputum

Gastroesophageal reflux disease with extra-esophageal symptoms, especially those with respiratory distress was attracting more and more attention. The related mechanisms were still in controversy. The purpose of the work was to explore airway inflammation triggered by gastroesophageal reflux.. Sixteen Sprague-Dawley rats were used as study group and 9 as control. In the study group, a plastic extender with a trumpet-shaped distal end was inserted into the lower esophagus to dilate the cardia, the pylorus was ligated. One ml of 0.1 mol/L hydrochloric acid was injected into the stomach. While a simple laparotomy was performed for control animals. All animals from two groups were sacrificed 24 hours after operation. Then tracheotomy was carried and the bronchoalveolar lavage fluid was collected in all animals. Cells in the fluid were counted and levels of interleukin (IL)-5, -6, -8 in it were measured.. Compared with control group, the study group presented a neutrophil pattern of airway inflammation and an elevated concentration of IL-5, -6, -8 with no significant difference regarding eosinophil count.. The gastroesophageal reflux-triggered airway inflammation is characterized by a neutrophilic airway inflammation which differed from that caused by asthma, and enhanced levels of IL-5, -6 and -8, which are similar to that caused by asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Gastroesophageal Reflux; Inflammation; Interleukin-5; Interleukin-6; Interleukin-8; Male; Rats; Rats, Sprague-Dawley

Although the clinical pictures of asthma and chronic obstructive pulmonary disease (COPD) may be similar, the pathogenesis differs in many aspects. The aim of the present study was to compare the cellular and biochemical features of airway inflammation in patients with asthma and COPD. The study was conducted in 22 patients with asthma (M/F 12/10, mean age 36 +/-14 years) and 17 patients with COPD (M/F 10/7, mean age 57 +/-11 years). Each patient underwent sputum induction followed by bronchoscopy, and bronchoalveolar lavage. Total and differential cell counts and the concentration of interleukin-8 (IL-8) and myeloperoxidase (MPO) were measured in induced sputum (IS) and BALF. We found no significant differences in the total and differential cell counts in IS between asthma and COPD patients. However, COPD patients showed an increased total macrophage count in BALF compared with asthma patients. The relative eosinophil count in BALF was significantly higher in patients with asthma vs. COPD. The concentration of IL-8 in IS and BALF was significantly higher in patients with COPD vs. asthma patients. The BALF concentration of MPO was significantly higher in patients with COPD compared with asthma patients. We conclude that the comparison of cellular composition and the concentration of inflammatory mediators in IS does not differentiate between asthma and COPD. The evaluation of BALF reveals more differences in the cellular and biochemical features of airways inflammation in patients with asthma and COPD than that of IS.

Topics: Adult; Aged; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Bronchoscopy; Female; Humans; Inflammation; Interleukin-8; Male; Middle Aged; Peroxidase; Pulmonary Disease, Chronic Obstructive; Respiratory Function Tests; Sputum

Some studies of severe asthma suggest that persistence or alteration in the pattern of inflammation may be associated with the severity of the disease. Whether there are differences in the expression of the principal cytokines and chemokines relevant to eosinophilic and neutrophilic inflammation in the airway tissues of severe compared to moderate asthmatics has not been determined. The aim of this study was to compare the patterns of expression of representative T-helper (Th) type 1 (interferon [IFN]-gamma) and Th-2 cytokines (interleukin [IL]-4, IL-5) and the neutrophil- and eosinophil-associated chemokines (IL-8 and eotaxin) in the airway tissues of patients with severe and moderate asthma.. Subjects with severe asthma (n = 24) and a comparison moderate asthma group (n = 26) were assessed using spirometry, induced sputum, exhaled nitric oxide, and bronchial biopsy. The expression of proteins of interest in the epithelium and subepithelium of the airway wall was examined by immunocytochemistry.. Subjects with severe asthma were more symptomatic, had a lower FEV(1), and had more sputum neutrophilia (p = 0.007) and eosinophilia (p = 0.001). Exhaled nitric oxide was similar between groups. IL-8 and IFN-gamma expression were increased and IL-4 expression was decreased in severe asthma compared to moderate disease (p < 0.001 for each comparison). Eotaxin and IL-5 expression did not differ between the groups.. Patients with severe asthma have increases in neutrophils and eosinophils in the sputum, and differ in airway cytokine/chemokine expression from moderate asthmatics. Excess neutrophilia may be explained by increased expression of IL-8, but differences in eosinophilia do not appear to be associated with IL-5 and eotaxin expression.

Topics: Adolescent; Adult; Aged; Asthma; Cytokines; Female; Humans; Immunohistochemistry; Interferon-gamma; Interleukin-8; Leukocyte Count; Male; Middle Aged; Neutrophils; Severity of Illness Index; T-Lymphocytes, Helper-Inducer

The CXC chemokines, IP-10/CXCL10 and IL-8/CXCL8, play a role in obstructive lung disease by attracting Th1/Tc1 lymphocytes and neutrophils, respectively. Inhaled corticosteroids (ICS) and long acting beta 2-agonists (LABA) are widely used. However, their effect(s) on the release of IP-10 and IL-8 by airway epithelial cells are poorly understood. This study examined the effects of fluticasone, salmeterol, and agents which raise intracellular cAMP (cilomilast and db-cAMP) on the expression of IP-10 and IL-8 protein and mRNA. Studies were performed in cultured human airway epithelial cells during cytokine-stimulated IP-10 and IL-8 release. Cytokine treatment (TNF-alpha, IL-1beta and IFN-gamma) increased IP-10 and IL-8 protein and mRNA levels. Fluticasone (0.1 nM to 1 microM) increased IP-10 but reduced IL-8 protein release without changing IP-10 mRNA levels assessed by real time RT-PCR. The combination of salmeterol (1 micro M) and cilomilast (1-10 mu M) reduced IP-10 but had no effect on IL-8 protein. Salmeterol alone (1 micro M) and db-cAMP alone (1 mM) antagonised the effects of fluticasone on IP-10 but not IL-8 protein. In human airway epithelial cells, inhibition by salmeterol of fluticasone-enhanced IP-10 release may be an important therapeutic effect of the LABA/ICS combination not present when the two drugs are used separately.

Topics: Adrenergic beta-Agonists; Albuterol; Asthma; Carboxylic Acids; Cells, Cultured; Chemokine CXCL10; Cyclohexanecarboxylic Acids; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Gene Expression; Humans; Interleukin-8; Nitriles; Phosphodiesterase Inhibitors; Respiratory Mucosa; Reverse Transcriptase Polymerase Chain Reaction; RNA; Salmeterol Xinafoate

Rhinovirus infection is the most common cause of acute exacerbations of inflammatory lung diseases, such as asthma and chronic obstructive pulmonary disease, where it provokes steroid refractory and abnormally intense neutrophilic inflammation that can be life threatening. Epidermal growth factor receptor (EGFR) expression correlates with disease severity and neutrophil infiltration in these conditions. However, the role of EGFR signaling in rhinovirus infection is unknown. We measured the key determinants of neutrophilic inflammation interleukin (IL)-8 and ICAM-1 in rhinovirus (RV16 serotype)-infected bronchial epithelial cells, BEAS-2B. RV16 infection stimulated IL-8 and ICAM-1 expression, which was further elevated (2-fold) by transient up-regulation of EGFR levels. Detection of viral RNA by quantitative real time PCR confirmed that enhanced expression was not associated with increased viral replication. EGFR ligands (epiregulin, amphiregulin, and heparin-binding epidermal growth factor) were induced by RV16 infection, and inhibition of metalloproteases responsible for ligand shedding partially suppressed this response. The EGFR inhibitor AG1478, completely blocked IL-8 and ICAM-1 expression to basal levels, as did the specific Erk1/2 inhibitor U0126. The p38 mitogen-activated protein kinase inhibitor SB203580 blocked IL-8 secretion but not ICAM-1 expression, whereas the PI3K inhibitor wortmannin was ineffective in both responses. Kinase inactive K721R EGFR, which is selectively deficient in STAT signaling, reversed RV16 responses associated with EGFR overexpression. In conclusion, RV16 infection rapidly promotes induction of EGFR ligands and utilizes EGFR signaling to increase IL-8 and ICAM-1 levels. These results suggest that targeting EGFR may provide a selective therapy that dampens neutrophil-driven inflammation without compromising essential antiviral pathways mediated by pathogen recognition receptors such as TLR3.

Topics: Amino Acid Substitution; Asthma; Common Cold; Enzyme Inhibitors; ErbB Receptors; Gene Expression Regulation; HeLa Cells; Humans; Inflammation; Intercellular Adhesion Molecule-1; Interleukin-8; Ligands; Matrix Metalloproteinases; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Neutrophils; Pulmonary Disease, Chronic Obstructive; Respiratory Mucosa; Rhinovirus; RNA, Viral; Signal Transduction; STAT Transcription Factors; Toll-Like Receptor 3

Mycoplasma pneumoniae is an extracellular pathogen, residing on mucosal surfaces of the respiratory and genital tracts. The lack of cell walls in mycoplasmas facilitates the direct contact of the bacterial membrane with the host cell. The cell membrane of mycoplasma is the major inducer of the host pathogenic response. Airway diseases caused by M. pneumoniae include bronchiolitis, bronchitis, and rarely bronchiectasis. In such disorders, neutrophil infiltration of the airways predominates. More recently, M. pneumoniae has been implicated in the pathogenesis of asthma. Epithelial cells play an important role in recruiting inflammatory cells into the airways. Since M. pneumoniae infection of human epithelial cells induces expression of IL-8-a potent activator of neutrophils-we investigated the signaling and transcriptional mechanisms by which mycoplasma membrane induces expression of this chemokine. In BEAS-2B human bronchial epithelial cells, mycoplasma membrane fraction (MMF) increased IL-8 mRNA and protein production. Activation of the transcriptional elements activating protein-1, nuclear factor-interleukin-6, and particularly NF-kappaB are essential for optimal IL-8 production by MMF. The mitogen-activated protein kinases individually played a modest role in MMF-induced IL-8 production. Toll-like receptor-2 did not play a significant role in MMF-induction of IL-8. Antibiotics with microbicidal activity against M. pneumoniae are also known to have anti-inflammatory effects. Whereas clarithromycin, azithromycin, and moxifloxacin individually were able to inhibit TNF-alpha-induction of IL-8, each failed to inhibit MMF-induction of IL-8.

Topics: Anti-Bacterial Agents; Asthma; Bronchi; Cell Membrane; Epithelial Cells; Extracellular Signal-Regulated MAP Kinases; Gene Expression Regulation; Humans; Interleukin-8; MAP Kinase Signaling System; Mycoplasma pneumoniae; Pneumonia, Mycoplasma; Toll-Like Receptor 2; Transcription Factors; Transcription, Genetic; Tumor Necrosis Factor-alpha

The role of the innate immune system in the pathogenesis of asthma is unclear. Activation of innate immune receptors in response to bacterial lipopolysaccharide, viral infection and particulate matter triggers a pre-programmed inflammatory response, which involves interleukin (IL)8 and neutrophil influx. The inflammatory response in asthma is heterogeneous.. To test the hypothesis that innate immune activation may be a relevant inflammatory mechanism in neutrophilic asthma where IL8 levels are increased.. Induced sputum was obtained from non-smoking adults with asthma (n = 49), healthy controls (n = 13) and a positive reference group with bronchiectasis (n = 9). Subjects with asthma were classified into inflammatory subtypes using induced sputum cell counts. Sputum was examined for mRNA expression of the innate immune receptors toll-like receptor (TLR)2, TLR4 and CD14, and inflammatory cytokines. A separate sputum portion was dispersed and the supernatant assayed for surfactant protein A, IL8, soluble CD14 and endotoxin.. Expression of innate immune receptors was increased in subjects with bronchiectasis and neutrophilic asthma compared with other asthma subtypes and controls. Increased expression of the receptors TLR2, TLR4 and CD14, as well as the pro-inflammatory cytokines IL8 and IL1beta, was observed. Subjects with neutrophilic asthma had higher airway levels of endotoxin than the other groups studied.. There is evidence of activation of the innate immune system in asthma which results in the production of pro-inflammatory cytokines and may contribute to the pathogenesis of neutrophilic asthma.

Topics: Adult; Aged; Asthma; Bronchiectasis; Cross-Sectional Studies; Endotoxins; Eosinophils; Female; Humans; Immunity, Cellular; Interleukin-8; Male; Middle Aged; RNA, Messenger; Toll-Like Receptors

The aim of the present study was to elucidate whether the culture of cells recovered from induced sputum may represent a suitable model to evaluate cytokine and chemokine production by airway inflammatory cells. Sputum induction was performed in 21 normal subjects and 30 asthmatic patients. A total of 21 out of the 30 asthmatic patients were taking inhaled corticosteroids, while the remaining nine were steroid-naive asthmatics. The steroid-naive group was evaluated before and after a 14-day treatment with oral prednisone (40 mg.day(-1)). The supernatant of lysed and centrifuged sputum and the supernatant of sputum cell culture were analysed. Tumour necrosis factor-alpha, interleukin (IL)-8 (CXCL8), IL-1beta, IL-13 and eotaxin-2 (CCL24) concentrations were determined by specific ELISA. Eotaxin-2 production by cell culture was higher in the asthma group (131+/-108 pg.mL(-1)) than in the control group (36+/-41 pg.mL(-1)) and treatment with oral corticosteroids eliminated this difference. In addition, reduction of eotaxin-2 levels by corticosteroid treatment was greater in cell culture (81.3% reduction) than in sputum (26.4%). There was correlation between the decrease in eotaxin-2 production and the decrease in blood eosinophil number and between eotaxin-2 and eosinophils in sputum. Eotaxin-2 may play an important role in asthma and the response to corticosteroid treatment suggests that analysis of sputum cell culture is relevant as an inflammatory parameter.

Topics: Administration, Inhalation; Administration, Oral; Adult; Anti-Inflammatory Agents; Asthma; Budesonide; Cells, Cultured; Chemokine CCL24; Chemokines; Chemokines, CC; Cytokines; Enzyme-Linked Immunosorbent Assay; Eosinophils; Female; Forced Expiratory Volume; Humans; Interleukin-13; Interleukin-1beta; Interleukin-8; Leukocyte Count; Lipopolysaccharides; Male; Middle Aged; Prednisone; Reference Values; Saliva; Statistics as Topic; Tumor Necrosis Factor-alpha

There is increasing evidence that both neutrophilic and eosinophilic inflammation persist in the airways of patients with severe asthma. We have reported a positive relationship between the concentrations of eosinophils and neutrophils in sputum from severe asthmatics, suggesting a possible role of eosinophils in regulating neutrophilic inflammation. The aim of this study was to investigate whether activated eosinophils modify the trans-basement membrane migration (TBM) of neutrophils.. Eosinophils and neutrophils were isolated from peripheral blood drawn from healthy donors. The TBM of neutrophils in response to a variety of chemoattractants was evaluated in the presence or absence of eosinophils by using the chambers with a Matrigel-coated Transwell insert.. As expected, eotaxin (10 nM) and RANTES (10 nM), but not IL-8 (10 nM), induced the TBM of eosinophils. On the contrary, only IL-8 induced the TBM of neutrophils. When eosinophils were coincubated with neutrophils and stimulated with IL-8, the TBM of eosinophils was significantly augmented. On the other hand, when neutrophils were coincubated with eosinophils and stimulated with eotaxin or RANTES, the TBM of neutrophils was not modified.. Neutrophils migrated by IL-8 may lead eosinophils to accumulate in the airways of patients with severe asthma. On the other hand, it is unlikely that eosinophils migrated by chemoattractants such as CC chemokines regulate neutrophilic inflammation.

Topics: Adult; Asthma; Basement Membrane; Cells, Cultured; Chemokine CCL11; Chemokine CCL5; Chemokines, CC; Chemotaxis, Leukocyte; Eosinophils; Female; Humans; In Vitro Techniques; Interleukin-8; Leukotriene B4; Male; Neutrophils; Pulmonary Eosinophilia

Airway inflammation is associated with an increased expression and release of inflammatory reactants that regulate processes of cell migration, activation and degranulation. The purpose of this study was to quantify bronchial lavage (BAL) fluid and serum levels of chemokine (IL-8), secretory leukocyte protease inhibitor (SLPI), soluble intracellular adhesion molecules-1 (sICAM-1) and sCD14, as surrogate markers of inflammatory and immune response in asthma and chronic obstructive pulmonary disease (COPD) patients with similar disease duration time.. Biomarkers in serum and BAL fluid from asthma (n=13) and COPD (n=25) patients were measured using commercially available ELISA kits.. We found that in asthma and COPD groups the concentrations of IL-8 and SLPI are significantly higher in BAL fluid than in serum, while levels of sICAM-1 and sCD14 in BAL fluid are significantly lower than in serum. Of these 4 measured biomarkers, only the BAL IL-8 was higher in COPD patients when compared to asthma (P<0.05). In both groups, BAL IL-8 correlated with SLPI (r=0.577, P<0.01 and r=0.589, P<0.05, respectively). In patients with COPD the BAL sICAM-1 correlated with sCD14 (r=0.576, P<0.01), while in asthma patients BAL sICAM-1 correlated with FEV(1)/FVC (r=0.418, P<0.01). Moreover, in asthma patients the serum SLPI correlated with sCD14 (r=0.688, P<0.01) and serum sICAM-1 negatively correlated with FEV(1)/FVC (r=-0.582, P<0.05).. Our findings point to the importance of selecting a correct biological fluid when analyzing specific biomarkers, and also show that of 4 measured biomarkers, only the BAL IL-8 was higher in COPD patients when compared to asthma.

Topics: Adult; Aged; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Bronchoscopy; Female; Forced Expiratory Volume; Humans; Inflammation Mediators; Intercellular Adhesion Molecule-1; Interleukin-8; Lipopolysaccharide Receptors; Male; Middle Aged; Pulmonary Disease, Chronic Obstructive; Secretory Leukocyte Peptidase Inhibitor; Vital Capacity

Rhinovirus-induced acute asthma is the most frequent trigger for asthma exacerbations.. We assessed which inflammatory mediators were released from bronchial epithelial cells (BECs) after infection with rhinovirus and then determined whether they were also present in subjects with acute virus-induced asthma, with the aim to identify a biomarker or biomarkers for acute virus-induced asthma.. BECs were obtained from bronchial brushings of steroid-naive asthmatic subjects and healthy nonatopic control subjects. Cells were infected with rhinovirus 16. Inflammatory mediators were measured by means of flow cytometry with a cytometric bead array. Subjects with acute asthma and virus infection were recruited; they were characterized clinically by using lung function tests and had blood taken to measure the inflammatory mediators identified as important by the BEC experiments.. IFN-gamma-induced protein 10 (IP-10) and RANTES were released in the greatest quantities, followed by IL-6, IL-8, and TNF-alpha. Dexamethasone treatment of BECs only partially suppressed IP-10 and TNF-alpha but was more effective at suppressing RANTES, IL-6, and IL-8. In acute clinical asthma serum IP-10 levels were increased to a greater extent in those with acute virus-induced asthma (median of 604 pg/mL compared with 167 pg/mL in those with non-virus-induced acute asthma, P < .01). Increased serum IP-10 levels were predictive of virus-induced asthma (odds ratio, 44.3 [95% CI, 3.9-100.3]). Increased serum IP-10 levels were strongly associated with more severe airflow obstruction (r = -0.8; P < .01).. IP-10 release is specific to acute virus-induced asthma.. Measurement of serum IP-10 could be used to predict a viral trigger to acute asthma.

Topics: Adolescent; Adult; Aged; Anti-Inflammatory Agents; Asthma; Biomarkers; Cells, Cultured; Chemokine CCL5; Chemokine CXCL10; Chemokines, CXC; Dexamethasone; Epithelial Cells; Flow Cytometry; Humans; Interleukin-6; Interleukin-8; Lung; Middle Aged; Picornaviridae Infections; Rhinovirus; Tumor Necrosis Factor-alpha

Cockroaches have been known as a cause of respiratory allergies such as asthma. IL-8 plays an integral role in the coordination and persistence of the inflammatory process in the chronic inflammation of the airways in asthma.. We investigated the mechanism by which German cockroach extract (GCE) triggers IL-8 release from human airway epithelial cells.. Chemical inhibitors were pretreated before addition of GCE for promoter activity and protein synthesis of IL-8. The Transcriptional activity of IL-8 promoter was analysed by mutational, deletional anaylsis and electrophoretic mobility shift assay (EMSA).. Stimulation of H292 cells with GCE resulted in a time- and concentration-dependent induction of IL-8 transcription and protein synthesis. IL-8 promoter deletion analysis indicated that position -132 to +41 was essential for GCE-induced IL-8 transcription, and mutants with substitutions in activator protein (AP)-1, nuclear factor (NF)-IL6 and NF-kappaB-binding sites revealed a requirement for NF-kappaB and NF-IL6, but not AP-1, in GCE-induced activation of the IL-8 promoter. The DNA-binding activities of NF-kappaB and NF-IL6 were induced by GCE, as determined by EMSA. The chemical inhibition of extracellular signal-regulated kinase (ERK) attenuated GCE-induced transcriptional activity and protein synthesis. In addition, through aprotinin treatment and PAR2 small interfering RNA transfection, it was proven that protease of GCE is consistent with the regulation of GCE-induced IL-8.. We conclude that GCE with protease activity-induced IL-8 expression is regulated by transcriptional activation of NF-kappaB and NF-IL6 coordinating with the ERK pathway in human airway epithelial cells.

Topics: Animals; Asthma; CCAAT-Enhancer-Binding Protein-beta; Cell Line, Tumor; Cockroaches; Extracellular Signal-Regulated MAP Kinases; Gene Expression Regulation; Humans; Interleukin-8; NF-kappa B; Respiratory Mucosa; Tissue Extracts

Interleukin 9 (IL-9) is a cytokine produced by activated T lymphocytes and that activates in vitro mast cells as well as T and B lymphocytes. In vivo, transgenic mice overexpressing the gene encoding IL-9 show several of the hallmarks of human allergic asthma: increased IgE concentration, bronchial mastocytosis, eosinophilia, increased mucus production, as well as bronchial hyperresponsiveness. Whereas some of these features reflect direct IL-9 activities on target cells such as mast cells and B lymphocytes, increased mucus production and eosinophilia rather result from IL-13 and IL-5 production induced by IL-9 in T lymphocytes and mast cells. Preclinical studies in mice have shown that anti-IL-9 blocking antibodies interfere with the development of asthma-like reactions. In the human species, asthmatic patients produce large amounts of this cytokine and IL-9 production correlates nicely with species biological parameters of the disease. Phase 2 clinical trials are in progress to test the efficacy of anti-IL-9 antibodies in humans.

Topics: Animals; Asthma; Bronchial Diseases; Bronchial Hyperreactivity; Cells, Cultured; Clinical Trials, Phase II as Topic; Disease Models, Animal; Eosinophilia; Humans; Immunoglobulin E; Interleukin-8; Mast Cells; Mastocytosis; Mice; Mice, Transgenic; T-Lymphocytes

Gastroesophageal reflux (GER) may induce respiratory symptoms (RS) through inhalation of acid gastric contents. To characterize the airway inflammation associated with this condition, 20 children [7.4 (0.9) yr old] with "difficult to treat" RS and a positive 24-h oesophageal pH monitoring (pHm) were studied and bronchoalveolar lavage (BAL) performed. The control group included 10 children [7.3 (1.3) yr], non-atopics, with a respiratory clinical history similar to the cases but no reflux, as demonstrated by a negative 24-h oesophageal pHm. On BAL samples, in addition to inflammatory indexes, the lipid-laden macrophage (LLM) index was determined as index of gastric content inhalation. As compared to controls, GER children had higher neutrophil proportion (P=0.002), higher LLM index (P=0.004) and higher concentrations of interleukin (IL)-8 (P=0.005), myeloperoxidase (MPO) (P=0.001) and elastase (P=0.045) in BAL fluid. In GER children, but not in controls, neutrophil proportion significantly correlated with LLM index (r=0.65, P=0.002), with IL-8 (r=0.62, P=0.003) and MPO levels (r=0.54, P=0.014) but not with elastase concentrations. These results suggest an active pathogenetic role of IL-8 in the recruitment and activation of neutrophils in the airways of children with GER, respiratory symptoms and BAL findings suggestive of gastric content aspiration.

Topics: Asthma; Bronchitis; Bronchoalveolar Lavage Fluid; Child; Enzyme-Linked Immunosorbent Assay; Female; Gastroesophageal Reflux; Humans; Hydrogen-Ion Concentration; Interleukin-8; Leukocytosis; Male; Neutrophil Activation; Neutrophils; Peroxidase; Respiratory Function Tests

The aim of this study was to investigate differences in airway inflammation between childhood and adult-onset asthma. A total of 47 asthmatic subjects were recruited from patients attending outpatient clinic. A group of 32 adults, mean age 42.8 years (yrs) and a group of 15 children, mean age 11.7 yrs were included. The two groups did not differ in respect to gender, dose of inhaled corticosteroids, atopy status or duration of asthma (mean duration 7.75 yr). Lung function tests, and sputum induction were performed. Flowcytometry was used to study cell population and interleukin-8, eosinophilic cationic protein (ECP) and granulocyte-macrophage colony stimulating factor were measured by enzyme-linked immunosorbent assay (ELISA). Three out of 15 (20%) of the children and 6 out of 32 (19%) of the adult patients were unable to produce a sufficient sputum sample. However, all individuals tolerated the procedure well. The viability of induced sputum cells did not differ among adult-onset asthmatics and children with asthma. Children had greater number of total cells in induced sputum compared with adult subjects (P=0.02). No statistical difference in T-lymphocytes subsets was found between the two groups, except for CD25 (P=0.04). A negative correlation was found between forced expiratory volume (FEV1) values and ECP levels (r=0.338, P=0.04) in the whole population (children and adults). Our study showed that the immunopathology of pediatric and adult asthma is similar and sputum induction provides opportunities for comparison of airway inflammation in childhood and adult asthma safely.

Topics: Adult; Age of Onset; Asthma; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Child; Enzyme-Linked Immunosorbent Assay; Eosinophil Cationic Protein; Female; Flow Cytometry; Forced Expiratory Volume; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Male; Sputum; T-Lymphocyte Subsets

Neutrophilic inflammation observed with severe asthma is often associated with interleukin-8 (IL-8). Neutrophils can secrete a variety of mediators that may augment the migration of eosinophils. We have reported a positive correlation between the concentrations of neutrophils and eosinophils in sputum from subjects with severe asthma, suggesting a possible role of neutrophils in regulating eosinophilic inflammation. The aim of this study was to investigate whether neutrophils stimulated with IL-8 modify the trans-basement membrane migration (TBM) of eosinophils. Eosinophils and neutrophils were isolated from peripheral blood drawn from healthy donors or subjects with mild asthma. The TBM of eosinophils in response to IL-8 was evaluated in the presence or absence of neutrophils using the chambers with a Matrigel-coated transwell insert. Neither IL-8 alone nor the presence of neutrophils alone induced the TBM of eosinophils. However, when eosinophils were coincubated with neutrophils and stimulated with IL-8, the TBM of eosinophils was significantly augmented. This augmented TBM of eosinophils was inhibited by a matrix metalloproteinase-9 inhibitor, a leukotriene B4 receptor antagonist, platelet-activating factor antagonists, or an anti-TNF-alpha monoclonal antibodies. These results suggest that neutrophils migrated in response to IL-8 may lead eosinophils to accumulate in the airways of asthma and possibly aggravate this disease.

Topics: Adult; Antibodies, Monoclonal; Asthma; Azepines; Basement Membrane; Chemotaxis, Leukocyte; Coculture Techniques; Collagen; Culture Media, Conditioned; Drug Combinations; Eosinophils; Humans; Interleukin-8; Laminin; Leukotriene B4; Matrix Metalloproteinase 9; Neutrophil Activation; Neutrophils; Paracrine Communication; Platelet Activating Factor; Protease Inhibitors; Proteoglycans; Pulmonary Eosinophilia; Triazoles; Tumor Necrosis Factor-alpha

We examined whether epithelial damage is associated with mobilization of neutrophils or eosinophils in the airway lumen during acute exacerbations of paediatric asthma. Aspirated sputum samples were harvested from 65 paediatric patients (mean age 3.4 +/- 0.4 years) during acute exacerbations of asthma. Patients with signs of infection were excluded. The presence of conglomerates of epithelial cells (i.e. "Creola bodies") in the aspirated sputum was utilized as a marker of epithelial damage. Among the paediatric asthma patients, 60% displayed Creola bodies (CrB+: n = 39) in their sputum samples whereas the remaining patients did not (CrB-: n = 26). CrB+ patients displayed more than a 20-fold increase in the concentration of the neutrophil-mobilizing cytokine interleukin (IL)-8 (pg/ml) and of the neutrophil product neutrophil elastase (NE, g/l), respectively, compared with CrB- patients (IL-8: 7468.2 +/- 1953.6 versus 347.9 +/- 72.6, P < 0.01; NE: 2072.4 +/- 419.0 versus 438.5 +/- 125.7, P < 0.01). Even though not statistically significant, a corresponding trend was observed for the relative number of sputum neutrophils. In contrast, the concentration of the eosinophil-mobilizing cytokine IL-5 and the esoinophil product ECP tended to be lower in CrB+ than in CrB- patients (P > 0.05). In conclusion, as indicated by the analysis of aspirated sputum, epithelial damage is associated with a locally enhanced chemotactic signal for and activity of neutrophils, but not eosinophils, during acute exacerbations of paediatric asthma. It remains to be determined whether these indirect signs of neutrophil mobilization in the airway lumen mirror an increased number of neutrophils in the surrounding airway tissue.

Topics: Acute Disease; Asthma; Bradykinin; Child, Preschool; Eosinophil Cationic Protein; Eosinophils; Epithelial Cells; Family Health; Female; Humans; Immunoglobulin E; Infant; Interleukin-5; Interleukin-8; Leukocyte Count; Leukocyte Elastase; Male; Neutrophils; Sputum; Tumor Necrosis Factor-alpha

Exacerbations of asthma are associated with viral respiratory tract infections, of which rhinoviruses (RV) are the predominant virus type. Airway smooth muscle is important in asthma pathogenesis, however little is known about the potential interaction of RV and human airway smooth muscle cells (HASM). We hypothesised that rhinovirus induction of inflammatory cytokine release from airway smooth muscle is augmented and differentially regulated in asthmatic compared to normal HASM cells.. HASM cells, isolated from either asthmatic or non-asthmatic subjects, were infected with rhinovirus. Cytokine production was assayed by ELISA, ICAM-1 cell surface expression was assessed by FACS, and the transcription regulation of IL-6 was measured by luciferase activity.. RV-induced IL-6 release was significantly greater in HASM cells derived from asthmatic subjects compared to non-asthmatic subjects. This response was RV specific, as 5% serum- induced IL-6 release was not different in the two cell types. Whilst serum stimulated IL-8 production in cells from both subject groups, RV induced IL-8 production in only asthmatic derived HASM cells. The transcriptional induction of IL-6 was differentially regulated via C/EBP in the asthmatic and NF-kappaB + AP-1 in the non-asthmatic HASM cells.. This study demonstrates augmentation and differential transcriptional regulation of RV specific innate immune response in HASM cells derived from asthmatic and non-asthmatics, and may give valuable insight into the mechanisms of RV-induced asthma exacerbations.

Topics: Adolescent; Adult; Asthma; CCAAT-Enhancer-Binding Protein-alpha; Cell Death; Cells, Cultured; Female; Humans; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; Male; Myocytes, Smooth Muscle; Respiratory System; Rhinovirus; Transcription, Genetic; Transfection

It has been suggested that patients with noneosinophilic asthma (NEA) show increased numbers of sputum neutrophils and a lack of response to therapy with corticosteroids, which are features that are commonly related to COPD. The aim of our study was to test the hypothesis that airway inflammation in NEA patients is different from that seen in patients with eosinophilic asthma (EA) and is similar to COPD.. Sputum cellular stress markers and neutrophilic and eosinophilic fluid-phase mediators were analyzed in asthma and COPD patients. NEA patients were identified based on a sputum eosinophil count of < or = 2.2% of the total nonsquamous cell count, and were compared to EA and COPD patients.. University Hospital of Heraklion, Department of Thoracic Medicine.. A total of 37 atopic asthmatic patients and 25 patients with COPD.. Sputum cell counts, cellular expression of heme oxygenase-1, inducible nitric oxide synthase, and nitrotyrosine, and sputum levels of eosinophilic cationic protein (ECP), myeloperoxidase (MPO), interleukin-8, and granulocyte macrophage colony-stimulating factor.. A total of 17 asthmatic patients (46%) belonged to the NEA group and 20 patients (54%) to the EA group. Patients with NEA showed no difference in neutrophil counts, fluid-phase mediators, or cellular stress markers compared to patients with EA. Compared to COPD patients, NEA patients showed the following significant differences: lower total cell counts (p < 0.03); lower neutrophil counts (p < 0.01); lower nitrotyrosine positive cell counts (p < 0.003); lower ECP levels (p < 0.005); lower MPO levels (p < 0.000); higher lymphocyte counts (p < 0.01); and higher macrophage counts (p < 0.03).. Despite low eosinophil counts, airway inflammation in NEA patients may share common features with that in EA patients but is distinct from COPD. Larger studies are needed to investigate further the clinical and inflammatory characteristics of NEA before we are able to categorize asthma patients into those with or without eosinophilic inflammation.

Topics: Aged; Asthma; Biomarkers; Cell Count; Eosinophil Cationic Protein; Eosinophils; Female; Forced Expiratory Volume; Humans; Immunohistochemistry; Inflammation; Interleukin-8; Male; Middle Aged; Oxidative Stress; Peroxidase; Pulmonary Disease, Chronic Obstructive; Severity of Illness Index; Sputum

Because cysteinyl-leukotrienes (cysLTs) are major protagonists in the pathophysiology of human asthma, and because neutrophils are involved in the more severe form of asthma, we studied the potential for leukotriene (LT) D(4) to induce synthesis of the chemokine IL-8 through activation of the CysLT1 receptor. We found LTD(4) to induce IL-8 gene expression in monocytic THP-1 cells and human dendritic cells with complete abrogation by selective CysLT1 antagonists. Human embryonic kidney-293 cells stably transfected with CysLT1 were used to better study the transcriptional regulation of the IL-8 promoter. Stimulation of the cells with graded concentrations of LTD(4) resulted in a time- and concentration-dependent induction of IL-8 transcription and protein synthesis. Use of IL-8 promoter mutants with substitutions in their NF-kappaB, activator protein (AP)-1, and NF-IL-6 binding elements revealed a requirement for NF-kappaB and AP-1, but not NF-IL-6, in LTD(4)-induced activation of the IL-8 promoter. Overexpression of dominant-negative IkappaBalpha inhibited the IL-8 transactivation induced by LTD(4). NF-kappaB DNA binding activity was induced by LTD(4), as determined by electrophoretic mobility shift assays, and could be supershifted by antibodies against p50 and p65. Supershift assays after LTD(4) stimulation also indicated the formation of a c-Jun/c-Fos complex. Moreover, our results demonstrate that LTD(4) upregulates the expression of c-fos and c-jun at the mRNA level. Our data show for the first time that LTD(4), via the CysLT1 receptor, can transcriptionally activate IL-8 production, with involvement of the transcription factors p50, p65, Fos, and Jun. These findings provide mechanistic and potentially therapeutic elements for modulation of the inflammatory component of asthma.

Topics: Asthma; Cell Line; Dendritic Cells; DNA; Dose-Response Relationship, Drug; Humans; Interleukin-8; Leukotriene D4; Membrane Proteins; Monocytes; Mutation; NF-kappa B p50 Subunit; Promoter Regions, Genetic; Protein Binding; Proto-Oncogene Proteins c-fos; Proto-Oncogene Proteins c-jun; Receptors, Leukotriene; RNA, Messenger; Time Factors; Transcription Factor AP-1; Transcription, Genetic; Transfection

Asthma rates have been increasing worldwide, and exposure to diesel exhaust particles (DEP) may be implicated in this increase. DEP may also play a role in the increased morbidity and mortality associated with ambient airborne particulate matter (PM) exposure. Two types of nasal responses have been reported for human subjects nasally instilled with one type of DEP: alterations in cytokines responses, and an increase in immunoglobulin E (IgE) production. Since DEP composition can vary depending on several factors, including fuel composition and engine load, the ability of another DEP particle and ozone-treated DEP to alter nasal IgE and cytokine production was examined. Nonasthmatic and asthmatic subjects were intranasally instilled with 300 microg NIST 1650 DEP per nostril, NIST 1650 DEP previously exposed to ozone (ozDEP; 300 microg/nostril), or vehicle. Subjects underwent nasal lavage before DEP exposure, and 4 and 96 h after exposure. Nasal cell populations and soluble mediators in the nasal lavage fluid were characterized. Total cell number, cell types, cell viability, concentrations of soluble mediators (including interleukin [IL]-8, IL-6, IgE, and granulocyte-macrophage colony-stimulating factor [GM-CSF]) were not altered by either DEP or ozDEP exposure. NO levels were not altered by either particle exposure. These findings suggest that DEP can be relatively noninflammatory and nontoxic, and that the physicochemical characteristics of DEP need to be considered when assessing the health effects of exposure to diesel exhaust.

Topics: Administration, Intranasal; Adolescent; Adult; Air Pollutants; Asthma; Cell Count; Cell Survival; Cytokines; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunoglobulin E; Interleukin-6; Interleukin-8; Male; Middle Aged; Nasal Lavage Fluid; Nasal Mucosa; Neutrophils; Ozone; Vehicle Emissions

Airway inflammation is a major factor in the pathogenesis of asthma. Interleukin 8 (IL8) is a potent proinflammatory cytokine that interacts with its receptors, IL8RA and IL8RB. We investigated the genetic polymorphisms in IL8, IL8RA, and IL8RB for any association with risk of asthma and peripheral blood eosinophil counts in a Korean population. By carrying out direct sequencing in 24 individuals, we identified 20 sequence variants within exons and their flanking regions, including the 1.5 kb promoter regions of IL8, IL8RA, and IL8RB. Among them, seven common single-nucleotide polymorphisms (SNPs) were selected for genotyping in our asthma cohort (n = 1,439). Two common haplotypes in IL8 and three in IL8RA and IL8RB (defined as one block) were identified. Although none of the polymorphisms showed a significant association with risk of asthma, IL8RA-B ht2 showed a significant association with the peripheral blood eosinophil counts (%) among asthma patients, e.g., lower eosinophil levels among individuals with the homozygous IL8RA-B ht2 (3.55 +/- 3.39%) than among other asthmatic patients (5.52 +/- 5.55%; P (corr) = 0.018). Our findings suggest that polymorphisms and haplotypes in IL8RA and IL8RB might be among the genetic factors underlying production of peripheral blood eosinophil.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Asthma; Case-Control Studies; Child; Child, Preschool; Eosinophils; Female; Genetic Variation; Humans; Interleukin-8; Korea; Leukocyte Count; Male; Middle Aged; Polymorphism, Single Nucleotide; Receptors, Interleukin-8A; Receptors, Interleukin-8B; Risk Factors

Staphylococcus aureus produces a set of proteins which act both as superantigens and toxins. Although their mode of action as superantigens is well understood, little is known about their effects on airway epithelial cells.. To investigate this problem, primary nasal epithelial cells derived from normal and asthmatic subjects were stimulated with staphylococcal enterotoxin A and B (SEA and SEB) and secreted (supernatants) and cell-associated (cell lysates) IL-8, TNF-alpha, RANTES and eotaxin were determined by specific ELISAs.. Non-toxic concentrations of SEA and SEB (0.01 microg/ml and 1.0 microg/ml) induced IL-8 secretion after 24 h of culture. Pre-treatment of the cells with IFN-gamma (50 IU/ml) resulted in a further increase of IL-8 secretion. In cells from healthy donors pretreated with IFN-gamma, SEA at 1.0 mug/ml induced release of 1009 pg/ml IL-8 (733.0-1216 pg/ml, median (range)) while in cells from asthmatic donors the same treatment induced significantly higher IL-8 secretion - 1550 pg/ml (1168.0-2000.0 pg/ml p = 0.04). Normal cells pre-treated with IFN-gamma and then cultured with SEB at 1.0 mug/ml released 904.6 pg/ml IL-8 (666.5-1169.0 pg/ml). Cells from asthmatics treated in the same way produced significantly higher amounts of IL-8--1665.0 pg/ml (1168.0-2000.0 pg/ml, p = 0.01). Blocking antibodies to MHC class II molecules added to cultures stimulated with SEA and SEB, reduced IL-8 secretion by about 40% in IFN-gamma unstimulated cultures and 75% in IFN-gamma stimulated cultures. No secretion of TNF-alpha, RANTES and eotaxin was noted.. Staphylococcal enterotoxins may have a role in the pathogenesis of asthma.

Topics: Adult; Asthma; Cells, Cultured; Enterotoxins; Female; Histocompatibility Antigens Class II; Humans; Interferon-gamma; Interleukin-8; Male; Nasal Mucosa; Staphylococcus aureus; Superantigens; Time Factors

Topics: Adult; Analysis of Variance; Asthma; Cell Differentiation; Cells, Cultured; Chemokine CCL5; Cytokines; Dexamethasone; Female; Humans; Interleukin-10; Interleukin-8; Male; Middle Aged; Sputum; Transforming Growth Factor beta; Transforming Growth Factor beta1

Severe asthma represents a heterogeneous group of patients whose characteristics of airway inflammation are poorly known.. To evaluate the sputum cytokine profiles of different phenotypes of severe asthma.. Severe asthmatic patients (n = 45) were divided into 3 groups: frequent exacerbations, persistent bronchoconstriction, and both features. Two other groups (9 patients with untreated mild asthma and 10 control subjects) were also studied. Selected sputum portions were assayed for differential cell count, supernatant interleukin 5 (IL-5), granulocyte-macrophage colony-stimulating factor, IL-8, and eosinophil cationic protein.. There were no statistically significant differences among the 3 severe asthma groups in terms of sputum inflammatory cell percentages, IL-8 levels, and eosinophil cationic protein levels, although IL-8 levels tended to be higher in patients with persistent bronchoconstriction. Sputum concentrations of granulocyte-macrophage colony-stimulating factor and IL-5 were significantly higher in patients with frequent exacerbations compared with the other 2 groups. Levels of IL-5 and IL-8 were higher in severe asthmatic patients compared with mild asthmatic patients and controls, whereas sputum eosinophil percentages were intermediate between those of mild asthmatic patients and controls.. Proeosinophilic cytokine levels are increased in severe asthmatic patients with frequent exacerbations but not in severe asthmatic patients with persistent bronchoconstriction, suggesting that different cytokine profiles could be associated with different phenotypes of severe asthma.

Topics: Adult; Aged; Asthma; Biomarkers; Cytokines; Eosinophils; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Inflammation; Interleukin-5; Interleukin-8; Male; Middle Aged; Neutrophils; Sputum

Severe asthma is characterized by elevated levels of pro-inflammatory cytokines and neutrophilic inflammation in the airways. Blood cytokines, markers of 'systemic' inflammation, may be a feature of amplified inflammation in severe asthma.. To detect differences in IL-8, TNF-alpha, IL-16 and IL-13 levels in the serum(s) of stable severe and mild-moderate asthmatics related to blood leucocytes proportion, airway calibre and exhaled nitric oxide (NO) levels.. We assessed cytokine serum levels by ELISA and blood leucocyte counts by an alkaline peroxidase method in 20 healthy controls, 22 mild-moderate [forced expiratory volume in 1 s (FEV1)(%pred): 89+/-3] and 14 severe asthmatics [FEV1(%pred): 49+/-2].. IL-8 and TNF-alpha levels were higher in severe asthmatics than in mild-moderate asthmatics or in controls (P<0.05). No differences in IL-16 and IL-13 levels were detected. Severe asthmatics showed higher circulating neutrophil and eosinophil number than controls (P<0.05). In severe asthmatics, exhaled NO levels were superior than in controls (P<0.05), but inferior than in mild-moderate asthmatics (P<0.05). We found positive correlation between TNF-alpha levels and exhaled NO (r=0.67; P=0.01) or circulating neutrophil counts (r=0.57; P=0.03) in severe asthmatics.. sTNF-alpha and sIL-8 are markers of 'systemic' inflammation in severe asthmatics, in conjunction with augmented circulating neutrophils, suggesting the involvement of neutrophil-derived cytokine pattern in severe asthma.

Topics: Adult; Analysis of Variance; Asthma; Biomarkers; Breath Tests; Case-Control Studies; Eosinophils; Female; Humans; Interleukin-13; Interleukin-16; Interleukin-8; Leukocyte Count; Male; Middle Aged; Neutrophils; Nitric Oxide; Respiratory Function Tests; Statistics, Nonparametric; Tumor Necrosis Factor-alpha

Lipoxins and 15-epi-lipoxins are lipid mediators that modulate leukocyte trafficking and promote the inflammation resolution. They are produced by different enzymatic pathways. Patients with severe asthma present ongoing airway inflammation despite chronic long-term treatment including oral glucocorticoids.. The aim of this study was to assess the presence of proinflammatory and anti-inflammatory mediators in the supernatants of induced sputum.. Induced sputum supernatants were collected from 10 normal subjects; 12 subjects with mild, 15 with moderate, and 24 with severe asthma; and 13 patients with chronic obstructive pulmonary disease. First, we validated the measurements of IL-8, leukotriene B 4 , lipoxin A 4 , and 15-epi-lipoxin A 4 in these samples. Then we measured these mediators by using immunoenzymatic methods.. IL-8 levels were highly increased in patients with severe asthma ( P < .0001), and leukotriene B 4 levels were significantly increased in patients with severe asthma and patients with chronic obstructive pulmonary disease. Lipoxin A 4 was significantly increased in the supernatant obtained from patients with mild asthma ( P < .0001), whereas 15-epi-lipoxin A 4 levels were higher in patients with severe asthma ( P = .05). More interestingly, we found a positive correlation between the level of lipoxin A 4 and IL-8 in patients with mild asthma.. These results indicate that induced sputum is a suitable method to assess lipoxin and 15-epi-lipoxin measurements in bronchi. The mechanisms involved in the synthesis of these 2 eicosanoid mediators would be helpful to understand better the imbalance between proinflammatory and anti-inflammatory mediators occurring in severe asthma. Lipoxin production involves interaction between lipoxygenases, whereas 15-epi-lipoxin production might involve CytP450 activity.

Topics: Adult; Asthma; Female; Humans; Interleukin-8; Leukotriene B4; Lipoxins; Male; Middle Aged; Pneumonia; Pulmonary Disease, Chronic Obstructive; Sputum

Several in vitro studies demonstrate that corticosteroids and long-acting beta(2) agonists may have a complementary and synergistic mode of action on the inflammatory processes in asthma.. Sputum was induced in 20 mild to moderate asthmatic patients and the induced sputum cells (ISC) were cultured with beclomethasone dipropionate (BDP) 10(-7) M, salbutamol 10(-8) M and formoterol 10(-8) M either alone or in combination, BDP plus salbutamol and BDP plus formoterol, for 24 h. We measured the levels of growth macrophages-colony stimulating factor (GM-CSF), released on activation normal T cells expressed and activated (RANTES) and interleukin-8 (IL-8), in the supernatant of stimulated cells by ELISA. Furthermore, we assessed nuclear translocation of glucocorticoid receptor (GR) and the expression of beta(2) receptor in ISC by immunofluorescence and RT-PCR, respectively.. The release of GM-CSF, RANTES and IL-8 in ISC was significantly reduced by BDP plus salbutamol or formoterol as compared with either drug alone (P < 0.0001). beta(2) receptor expression was increased after 30 min of incubation with BDP and continued to increase over a time period of 4 h (P < 0.0001). Furthermore after 30 min of incubation, nuclear translocation of GR was greater with BDP plus salbutamol or formoterol than with any of the drugs alone (P < 0.0001).. The present ex vivo study demonstrates a complementary mode of action between BDP and salbutamol or formoterol leading to an enhanced anti-inflammatory activity.

Topics: Adult; Albuterol; Anti-Asthmatic Agents; Asthma; Beclomethasone; Bronchodilator Agents; Cells, Cultured; Chemokine CCL5; Drug Interactions; Drug Therapy, Combination; Ethanolamines; Female; Formoterol Fumarate; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Male; Middle Aged; Receptors, Adrenergic, beta-2; Receptors, Glucocorticoid; Severity of Illness Index; Sputum; Tissue Distribution

Cytokines and chemokines have been implicated in the pathogenesis of asthma. Inhaled corticosteroids have been shown to decrease the number of eosinophils and to downregulate T H 2 cytokines but to increase neutrophils. The effect of corticosteroids on chemokine expression in asthma has not yet been investigated.. We sought to investigate the effect of a 2-week course of oral corticosteroid (methylprednisolone, 40 mg/d) on the expression of CXC chemokines (IL-8 and IFN-gamma-inducible protein 10 [IP-10]) and CC chemokines (eotaxin and monocyte chemotactic proteins [MCPs] 1-4) in endoscopic biopsy specimens of 13 patients with moderate-to-severe asthma.. CD3, major basic protein, and elastase immunoreactivities were monitored before and after treatment by using immunocytochemistry. Eotaxin, IL-8, IP-10, MCP-1, MCP-2, MCP-3, and MCP-4 mRNA expression in epithelium and submucosa were studied by using in situ hybridization.. Corticosteroids reduced the number of CD3-positive T cells and major basic protein-positive eosinophils ( P < .05), whereas the number of neutrophils were increased ( P < .05). Corticosteroids also reduced the number of eotaxin ( P < .05), MCP-3, and MCP-4 mRNA-positive cells ( P < .001) in the epithelium and subepithelium. However, corticosteroids caused a significant increase in the epithelial expression of IL-8 ( P < .001), IP-10 ( P < .05), and MCP-2 mRNAs ( P < .01). Corticosteroids had no effects on MCP-1 mRNA expression.. Our results demonstrate the dual nature of corticosteroids. Although corticosteroids can downregulate the expression of some asthma-associated chemokines, such as eotaxin, MCP-3, and MCP-4, they can also upregulate the expression of other chemokines, including IL-8, IP-10, and MCP-2. The failure of oral corticosteroids to inhibit IL-8 mRNA expression might contribute to persistent airway neutrophilia observed in patients with moderate-to-severe asthma, despite treatment with corticosteroids.

Topics: Administration, Oral; Adult; Asthma; Chemokine CXCL10; Chemokines; Chemokines, CC; Chemokines, CXC; Drug Administration Schedule; Eosinophils; Female; Glucocorticoids; Humans; Interleukin-8; Leukocyte Count; Male; Methylprednisolone; Neutrophils; Respiratory Mucosa; RNA, Messenger

The influx of leukocytes (eosinophils, lymphocytes, and monocytes) into the airways and their production of proinflammatory cytokines contribute to the severity of allergic asthma. We describe here the synthesis and pharmacological evaluation of a series of triazinylphenylalkylthiazolecarboxylic acid esters that were designed to act as lung-specific antedrugs and inhibitors of the production of interleukin (IL)-5, a primary eosinophil-activating and proinflammatory cytokine. Closer examination of the hydroxypropyl ester, 15, indicated its high metabolic stability (t(1/2) > 240 min) in human lung S9 fraction but rapid conversion (t(1/2) = 15 min) into the pharmacologically inactive carboxylic acid by human liver preparations. In stimulated human whole blood cultures, 15 reduced not only the production of IL-5 (IC(50) = 78 nM) but also the biosynthesis of the monocyte chemotactic proteins MCP-1 (IC(50) = 220 nM), MCP-2 (IC(50) = 580 nM), and MCP-3 (IC(50) = 80 nM). In vivo, intratracheal administration of 15 (6 mg/animal) to allergic sheep, either before (-4 h) or after (+1.5 h) the pulmonary allergen challenge, completely abrogated the late-phase airway response and reduced the bronchial hyperreactivity to inhaled carbachol.

Topics: Adult; Animals; Asthma; Bronchodilator Agents; Chemokine CCL2; Chemokine CCL7; Chemokine CCL8; Cytokines; Esters; Humans; In Vitro Techniques; Interleukin-4; Interleukin-5; Interleukin-8; Liver; Lung; Monocyte Chemoattractant Proteins; Sheep; Thiazoles; Triazines

Neutrophils are a major component of the inflammatory response in patients with asthma and other inflammatory conditions. Proinflammatory cytokines, such as tumor necrosis factor alpha (TNF-alpha), are increased in the airway of patients with severe asthma and have been implicated in the recruitment of neutrophils into areas of inflammation. Here, we show that TNF-alpha induces a stop signal that promotes firm neutrophil adhesion and inhibits neutrophil polarization and chemotaxis to chemoattractants including interleukin-8 and C5a. TNF-alpha treatment of neutrophils plated on a fibrinogen-coated surface promotes firm neutrophil adhesion and the formation of vinculin-containing focal complexes. TNF-alpha induces a more than tenfold increase in p38 mitogen-activated protein kinase (MAPK) but not extracellular signal-regulated kinase phosphorylation. Inhibition of p38 MAPK in neutrophils treated with TNF-alpha causes neutrophil polarization and motility. These findings suggest that TNF-alpha initiates a stop signal through a p38 MAPK pathway, which may promote the retention of neutrophils in inflammatory sites. Together, our data suggest that inhibition of p38 MAPK may be an attractive target to limit inflammatory responses that are mediated by TNF-alpha.

Topics: Asthma; Cell Adhesion; Cell Polarity; Cells, Cultured; Chemotaxis, Leukocyte; Complement C5a; Enzyme Inhibitors; Humans; Inflammation; Interleukin-8; MAP Kinase Signaling System; Neutrophils; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Signal Transduction; Tumor Necrosis Factor-alpha; Up-Regulation

Lung epithelial cells contribute to local inflammation by the production of pro-inflammatory mediators like interleukin (IL)-8 and IL-6. Although their production depends on gene transcription, previous studies showed that post-transcriptional mechanisms modulate IL-8 and IL-6 production. Human lung epithelial cells turn from normoresponsive into hyperresponsive IL-8- and IL-6-producing cells when their IL-8 and IL-6 mRNA degradation is reduced. We hypothesized that IL-17, a mediator predominantly released by memory T cells and present in airways of individuals with asthma, would modulate rather than induce IL-8 and IL-6 production by both human lung epithelial cells and fibroblasts. We show here for both cell types that IL-17 was a weak stimulus of IL-8 and IL-6 production, but markedly enhanced IL-8 and IL-6 responses to another stimulus, such as tumor necrosis factor-alpha. This modulatory effect of IL-17 was paralleled by a reduced IL-8 and IL-6 mRNA degradation, with no effect on IL-8 and IL-6 gene transcription. In conclusion, IL-17 particularly affects post-transcriptional regulation of IL-8 and IL-6 expression leading to enhanced IL-8 and IL-6 responses to secondary stimuli, and is only a weak proinflammatory stimulus by itself. This poses the interesting concept that by releasing IL-17 from memory T cells, the adaptive immune system instructs lung structural cells as part of the innate immune system to respond more vigorously.

Topics: Asthma; Cell Line; Cell Line, Tumor; Cell Nucleus; Chloramphenicol O-Acetyltransferase; Dose-Response Relationship, Drug; Epithelial Cells; Fibroblasts; Humans; Immune System; Interleukin-17; Interleukin-6; Interleukin-8; Lipopolysaccharides; Lung; Promoter Regions, Genetic; Respiratory Hypersensitivity; RNA Processing, Post-Transcriptional; RNA, Messenger; Time Factors; Transcription, Genetic; Transfection; Tumor Necrosis Factor-alpha

In this study, we measured the levels of eosinophil cationic protein (ECP) and interleukin (IL)-8 in bronchoalveolar lavage (BAL) fluid from patients with acute asthma and acute bronchiolitis, to determine any similarities or dissimilarities in the profiles of these biochemical markers in the two diseases.. BAL fluids were obtained from children with acute asthma (n=16), infants with acute bronchiolitis caused by respiratory syncytial virus (n=18), and control subjects (n=14). Children with asthma were selected to be free of viral infection. BAL cell counts and differentials were determined, and ECP and IL-8 levels were measured by radioimmunoassay and ELISA, respectively.. ECP levels in BAL fluids were significantly higher in the asthma group than in the bronchiolitis (P<0.01) or control (P<0.0001) groups. However, IL-8 levels were significantly higher in the bronchiolitis group than in the asthma (P<0.01) or control (P<0.001) groups. IL-8 levels in the asthma group and ECP levels in the bronchiolitis group were similar to those of the control group.. This difference in profiles of ECP and IL-8 in acute asthma and acute bronchiolitis, together with a different inflammatory cell pattern, suggests that the nature of the inflammatory process within the lower respiratory tract may be distinctive in these two diseases.

Topics: Acute Disease; Asthma; Bronchiolitis; Bronchoalveolar Lavage Fluid; Bronchoscopy; Child, Preschool; Eosinophil Cationic Protein; Eosinophils; Female; Humans; Interleukin-8; Macrophages; Male; Neutrophils

Asthma is characterized by both chronic inflammation and remodeling of the airways. Proteases are important mediators of inflammation, cytokine activation, and tissue remodeling.. This study investigated matrix metalloproteinase-9 (MMP-9) and neutrophil elastase (NE) enzyme activity in the sputum of subjects with different inflammatory phenotypes of asthma (eosinophilic, neutrophilic, and paucigranulocytic asthma) and in healthy control subjects.. Nonsmoking adults with asthma and healthy control subjects underwent hypertonic saline challenge and sputum induction. Selected sputum portions were dispersed with dithiothreitol and assayed for MMP-9 and NE enzyme activity.. Subjects with eosinophilic asthma had significantly more active MMP-9 (39 ng/ml) compared with those with neutrophilic asthma (10 ng/ml) and control subjects (2.5 ng/ml, p < 0.01). Although there were high levels of total MMP-9 in neutrophilic asthma (5,273 ng/ml), most (> 99%) was inactivated (and bound to tissue inhibitor of metalloproteinase-1). In neutrophilic asthma, more subjects had NE activity (39%) compared with both healthy control subjects (0%), subjects with eosinophilic asthma (6%), or subjects with paucigranulocytic asthma (0%, p < 0.05). There were strong and consistent positive correlations between interleukin-8, neutrophils, and proteolytic enzymes. MMP-9 was inversely correlated with NE (r = -0.93).. Proteolytic enzyme activity in asthma is dependent on the underlying inflammatory phenotype and is differentially regulated with MMP-9 activity a feature of eosinophilic inflammation, and active NE in neutrophilic inflammation.

Topics: Adolescent; Adult; Aged; Asthma; Eosinophils; Humans; Interleukin-8; Leukocyte Elastase; Male; Matrix Metalloproteinase 9; Middle Aged; Neutrophils; Peptide Hydrolases

IL-1beta may contribute to airway inflammation by inducing pro-inflammatory cytokines and chemokines from bronchial epithelial cells. In the current study, we investigated the cis-acting sites within the IL-8 promoter, and signalling pathways important in IL-8 production from BEAS2B cells following IL-1beta stimulation. IL-1beta treatment (0.1-10 ng/mL) upregulated IL-8 protein production in a dose dependent manner and IL-8 mRNA in a time dependent manner. IL-1beta induced upregulation of IL-8 promoter-reporter constructs, indicating that the mechanism of upregulation was pre-transcriptional. Using IL-8 promoter constructs with mutated cis-acting sites, it was found that both the NF-kappaB and NF-IL6 sites together were required for IL-8 promoter induction following IL-1beta treatment. Using chemical inhibitors or dominant negative mutants, we found that IL-8 promoter activity required IkappaB kinase beta, IkappaB, but not the MAP kinases p38 or c-Jun N-terminal kinase 2. Fluticasone propionate was able to suppress IL-1beta induced IL-8 protein and promoter activation, using both a -1481 bp fragment and a -133 bp fragment, indicating that the glucocorticoid response element found at -330 bp was not required for fluticasone mediated suppression of IL-8 promoter activation.

Topics: Androstadienes; Anti-Inflammatory Agents; Asthma; Bronchi; CCAAT-Enhancer-Binding Protein-beta; Cells, Cultured; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Fluticasone; Humans; Interleukin-1; Interleukin-8; NF-kappa B; Promoter Regions, Genetic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Transcription Factors; Up-Regulation

Topics: Adult; Asthma; Budesonide; Cell Differentiation; Eosinophil Cationic Protein; Female; Humans; Interleukin-17; Interleukin-8; Male; Middle Aged; Neutrophils; Peroxidase; Prospective Studies; Sputum

Nuclear factor (NF)-kappaB is a transcription factor known to regulate the expression of many inflammatory genes, including cytokines, chemokines, and adhesion molecules. NF-kappaB is held inactive in the cytoplasm, bound to I-kappaB. The removal of I-kappaB, via the actions of inhibitor of kappaB (I-kappaB) kinase-2 (IKK-2), allows NF-kappaB to enter the nucleus.. To determine the impact of inhibiting IKK-2 on in vitro and in vivo models of airway inflammation.. The effect of inhibiting IKK-2 was assessed in stimulated, cultured, primary human airway smooth muscle cells and an antigen-driven rat model of lung inflammation.. The release of cytokines from cultured cells and inflammatory cytokine expression and cellular burden in the lung were determined.. Two structurally distinct molecules and dominant negative technology demonstrated that inhibition of IKK-2 activity completely blocked cytokine release from cultured cells, whereas the two glucocorticoid comparators had limited impact on granulocyte colony-stimulating factor, interleukin 8, and eotaxin release. In addition, in an in vivo antigen-driven model of airway inflammation, the IKK-2 inhibitor blocked NF-kappaB nuclear translocation, which was associated with a reduction in inflammatory cytokine gene and protein expression, airway eosinophilia, and late asthmatic reaction, similar in magnitude to that obtained with budesonide.. This study demonstrates that inhibiting IKK-2 results in a general reduction of the inflammatory response in vitro and in vivo. Compounds of this class could have therapeutic utility in the treatment of asthma and may, in certain respects, possess a beneficial efficacy profile compared with that of a steroid.

Topics: Amides; Animals; Anti-Inflammatory Agents; Asthma; Budesonide; Cells, Cultured; Chemokine CCL11; Chemokines, CC; Dexamethasone; Disease Models, Animal; Drug Evaluation, Preclinical; Gene Expression; Granulocyte Colony-Stimulating Factor; Humans; I-kappa B Kinase; Inflammation; Interleukin-8; Muscle, Smooth; NF-kappa B; Rats; Respiratory System; Thiophenes

Airway remodeling in asthma comprises a range of structural changes. Several studies have suggested an association between these changes and disease severity. The relationship between the extent of remodeling and lung function is not well defined.. We sought to contrast the structural changes in the airways of well-defined groups of subjects with severe and moderate asthma and to correlate the extent of remodeling with disease severity.. Endobronchial biopsy specimens were obtained from 15 subjects with severe and 13 subjects with moderate asthma. Epithelial integrity, cell-layer areas, subepithelial fibrosis, and the distance between epithelial and airway smooth muscle (ASM) layers were measured by means of image analysis. Collagen was identified by using Van Giesen stain, and ASM was defined by using smooth muscle alpha-actin immunostaining. Specific immunostains were performed for the evaluation of RANTES, IL-8, and eotaxin expression as markers of ASM phenotype.. ASM area was greater in subjects with severe (0.24+/- 0.03 mm(2)) than in subjects with moderate (0.05+/- 0.01 mm(2)) asthma (P<.001). The distance between the epithelial and ASM layers was less in the severe group (0.12+/- 0.01 mm) than in the moderate group (0.24+/- 0.02, P<.001). A trend toward greater subepithelial fibrosis in subjects with severe asthma did not reach statistical significance. IL-8 and eotaxin expression, but not RANTES expression, were increased in the ASM of subjects with severe asthma compared with in subjects with moderate asthma.. Smooth muscle alteration is the key structural change that distinguishes severe from moderate asthma, and phenotypic change in ASM might contribute to the difficulty in obtaining adequate control in some subjects with severe asthma.

Topics: Adult; Asthma; Cell Size; Chemokine CCL11; Chemokine CCL5; Chemokines, CC; Female; Fibrosis; Forced Expiratory Volume; Humans; Image Processing, Computer-Assisted; Interleukin-8; Lung; Male; Middle Aged; Muscle, Smooth; Respiratory Mucosa

Airway inflammation is assessed to monitor progression, control and treatment of asthma. The collection of exhaled breath condensate (EBC) provides a non-invasive alternative to induced sputum samples for the monitoring of airway inflammation. Both samples can be confounded by salivary contamination. The aim of this study was to compare the levels of inflammatory mediators in samples of EBC, induced sputum and saliva samples from subjects with asthma.. EBC, saliva and induced sputum samples were collected from subjects with asthma (n=10). Total protein, IL-8, 8-isoprostane and surfactant protein A (SPA) were assessed in each sample.. Total protein, IL-8, 8-isoprostane and SPA were detected in all sputum samples. Only total protein and SPA were consistently measured in EBC, with levels at least 100-fold lower than those measured in induced sputum. In saliva, total protein, SPA and 8-isoprostane were detected in all samples, with IL-8 detected in 60% of samples.. Induced sputum is a reliable technique that can be used to assess markers of airway inflammation. While EBC is a simple and inexpensive technique to collect lower airway secretions, the detection of inflammatory mediators is variable, and further work is required to validate this technique to assess inflammatory mediators.

Topics: Adult; Aged; Asthma; Breath Tests; Bronchial Provocation Tests; Dinoprost; Female; Humans; Interleukin-8; Male; Middle Aged; Predictive Value of Tests; Pulmonary Surfactant-Associated Protein A; Saliva; Sputum; Statistics, Nonparametric

The levels of serum TNF-alpha and IL-8 in the patients with allergic asthma during acute attack period and remission period, and the effects of glucocorticoid (GC) on them were investigated. By using ELISA, the levels of TNF-alpha and IL-8 were detected in the healthy volunteers (group C, n =40), the patients with allergic asthma (n=40) during acute attack period (group A) and remission period (group B) and those taking GC for a week (n=28). The results were compared among them. It was found that the levels of TNF-alpha and IL-8 in group A were higher than in group B and group C. In the patients subject to GC therapy, the levels of TNF-alpha and IL-8 were decreased as compared with those in group A. In group B, the level of TNF-alpha was higher than in group C, but there was no significant difference in the level of IL-8 between group B and group C. It was concluded that the inflammatory cytokines, TNF-alpha and IL-8, played important roles in the bronchus allergic inflammation. GC could reduce the levels of serum TNF-alpha and IL-8 to exert the anti-inflammatory effects.

Topics: Adolescent; Adult; Aged; Anti-Inflammatory Agents; Asthma; Female; Humans; Interleukin-8; Male; Middle Aged; Prednisone; Tumor Necrosis Factor-alpha

Subjects with occupational asthma (OA) generally present asthma symptoms and airway hyperresponsiveness after cessation of exposure. We hypothesized that they are also left with airway inflammation. We assessed 133 subjects with OA at a mean interval of 8.7 years (0.5-20.8 years) after cessation of exposure by questionnaire, airway caliber, and responsiveness to methacholine. Satisfactory samples of induced sputum were obtained from 98 subjects. We defined three groups of subjects: (1) cured: normalization of the concentration of methacholine provoking a 20% decrease in FEV1 (PC20), (2) improved: increase in PC20 by 3.2-fold or more but PC20 still abnormal, and (3) not improved: no significant change in PC20. In all, 9/28 subjects (32.1%) with no improvement versus 6/56 (10.7%) subjects with partial and complete improvements had sputum eosinophils equal to or greater than 2% and 11/28 (39.3%) subjects versus 11/56 (19.6%) subjects showed sputum neutrophils equal to or greater than 61%. Levels of interleukin-8 and of the neutrophil-derived myeloperoxidase were significantly more elevated in sputum of subjects with no improvement. Those in the cured or improved groups had a significantly longer time lapse since diagnosis and a higher PC20 at the time of diagnosis. We conclude that failure to improve after cessation of exposure to an agent causing OA is associated with airway inflammation at follow-up.

Topics: Adult; Airway Resistance; Asthma; Bronchial Provocation Tests; Cohort Studies; Female; Follow-Up Studies; Humans; Inflammation; Inflammation Mediators; Interleukin-8; Logistic Models; Male; Methacholine Chloride; Middle Aged; Occupational Diseases; Occupational Exposure; Peroxidase; Probability; Respiratory Function Tests; Risk Assessment; Sampling Studies; Sputum; Time Factors

Secretory IgA contributes to humoral defense mechanisms against pathogens targeting mucosal surfaces, and secretory component (SC) fulfills multiple roles in this defense. The aims of this study were to quantify total SC and to analyze the form of free SC in sputa from normal subjects, subjects with asthma, and subjects with cystic fibrosis (CF). Significantly higher levels of SC were detected in CF compared with both other groups. Gel filtration chromatography revealed that SC in CF was relatively degraded. Free SC normally binds interleukin (IL)-8 and inhibits its function. However, in CF sputa, IL-8 binding to intact SC was reduced. Analysis of the total carbohydrate content of free SC signified overglycosylation in CF compared with normal subjects and subjects with asthma. Monosaccharide composition analysis of free SC from CF subjects revealed overfucosylation and undersialylation, in agreement with the reported CF glycosylation phenotype. SC binding to IL-8 did not interfere with the binding of IL-8 to heparin, indicating distinct binding sites on IL-8 for negative regulation of function by SC and heparin. We suggest that defective structure and function of SC contribute to the characteristic sustained inflammatory response in the CF airways.

Topics: Adult; Asthma; Biomarkers; Child; Cystic Fibrosis; Female; Glycosylation; Humans; Interleukin-8; Male; Phenotype; Probability; Prognosis; Reference Values; Respiratory Mucosa; Secretory Component; Sensitivity and Specificity; Severity of Illness Index; Sputum

Interleukin (IL)-10 is a pleiotropic cytokine with a broad spectrum of immunosuppressive and anti-inflammatory effects. IL-10 secretion from alveolar macrophages is defective in patients with asthma and lower concentrations of IL-10 are found in bronchoalveolar lavage (BAL) from asthmatic patients than in normal control subjects. Reduced IL-10 may result in exaggerated and more prolonged inflammatory responses in asthmatic airways. IL-10 acting through the IL-10 receptor (IL-10R) stimulates the transcription factors STAT1 and STAT3.. We investigated IL-10 and IL-10R expression in normal and asthmatic bronchial epithelium and BAL macrophages using reverse transcription-polymerase chain reaction, immunohistochemistry and Western blotting. The functional effect of IL-10 was examined using granulocyte-macrophage-colony stimulating factor, enzyme-linked immunosorbent assay and Western blotting for phosphorylated STAT1 and STAT3.. IL-10 was not expressed in epithelial cells; furthermore these cells did not express the IL-10R and had no functional response to exogenous IL-10. Bronchial epithelial cells expressed variable levels of phosphorylated STAT1 and STAT3 with no change in expression between normal subjects and asthmatics. IL-10 protein and IL-10R expression was detected in alveolar macrophages from all subjects.. Our study suggests that the bronchial epithelium is not a source of IL-10 and cannot respond to exogenous IL-10 because of a lack of IL-10R expression.

Topics: Adult; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Case-Control Studies; DNA-Binding Proteins; Epithelial Cells; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-10; Interleukin-8; Macrophages, Alveolar; Male; Receptors, Interleukin; Receptors, Interleukin-10; STAT1 Transcription Factor; STAT2 Transcription Factor; Trans-Activators

Excessive salivary contamination of induced sputum samples prevents the satisfactory examination of lower airway inflammation. The effects of salivary contamination on different sputum fluid phase measures and the levels of salivary contamination preventing analysis are not defined. The present study sought to examine this by investigating the effect of increasing salivary contamination on induced sputum samples. Sputum and saliva samples from subjects with asthma and healthy controls were collected, and treated with dithiothreitol (DTT). Saliva was then added to aliquots of dispersed sputum in increasing proportions (0% to 100%). The effect of increasing saliva contamination was assessed on sputum total cell count, viability, differential cell count and fluid phase levels of interleukin (IL)-8, eosinophil cationic protein (ECP) and total protein. The addition of saliva to induced sputum reduced total cell counts and absolute cell counts but did not change the differential cell count. Levels of fluid phase ECP and IL-8 were significantly reduced with increased salivary contamination. There was a progressive reduction in ECP and IL-8, which reached significance at 70% and 80% saliva contamination, respectively. IL-8 levels corrected for total protein showed no change with increasing saliva concentrations. Induced sputum differential cell counts expressed as the proportion of nonsquamous cells are robust measures that are not influenced by salivary contamination. Studies reporting total and absolute cell counts and fluid phase mediator levels require control for squamous contamination.

Topics: Adult; Aged; Asthma; Blood Proteins; Case-Control Studies; Cell Count; Eosinophil Granule Proteins; Female; Humans; Interleukin-8; Male; Middle Aged; Osmolar Concentration; Ribonucleases; Saliva; Sputum

Exhaled markers of airway inflammation become increasingly important in the management of childhood asthma. The aims of the present study are: 1) to compare exhaled markers of inflammation (nitric oxide, carbon monoxide, and acidity of breath condensate) with conventional asthma measures (lung function tests and asthma control score) in childhood asthma; and 2) to investigate the detectability of albumin, CRP, IL-6, IL-8, TNF-alpha, sICAM-1, and sTNF-R75 in the exhaled breath condensate (EBC) of asthmatic children. Thirty-two children with mild to moderate persistent asthma and healthy controls aged 6-12 years were studied. We measured exhaled NO and CO, and subsequently EBC was collected. Inflammatory mediators in EBC were measured using an enzyme-linked immunosorbent assay. Respiratory symptoms and asthma control were assessed using the asthma control questionnaire (ACQ) of Juniper et al. (Eur Respir J 1999;14:902-907). Exhaled NO showed a significant correlation with exhaled CO (r = 0.59, P < 0.05) and FEV1 (r = -0.59, P < 0.05), but not with ACQ score (r = 0.48, P = 0.06). Exhaled CO was correlated with prebronchodilator FEV1 (r = -0.45, P < 0.05), but not with asthma control (r = 0.18, P = 0.35). Acidity of EBC was significantly lower in asthmatic children than in healthy controls (P < 0.05), but did not correlate with any of the conventional asthma measures. We were not able to demonstrate the presence of CRP, IL-6, IL-8, TNF-alpha, sICAM-1, and sTNF-R75 in EBC. Albumin was found in two EBC samples of asthmatic children. We conclude that exhaled NO had a better correlation with lung function parameters and asthma control than exhaled CO and acidity of EBC, in mild to moderate persistent childhood asthma. However, exhaled NO, CO, and deaerated pH of EBC did not differ between asthmatic children and controls, possibly because of a too homogeneous and well-controlled study population. To further evaluate the clinical utility of exhaled markers in monitoring childhood asthma, more studies are required on a wider range of asthma severity, and preferably with repeated measurements of markers and of asthma control.

Topics: Adolescent; Albumins; Asthma; Breath Tests; C-Reactive Protein; Carbon Monoxide; Case-Control Studies; Child; Cytokines; Enzyme-Linked Immunosorbent Assay; Exhalation; Humans; Hydrogen-Ion Concentration; Inflammation Mediators; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; Nitric Oxide; Patient Selection; Receptors, Tumor Necrosis Factor, Type II; Respiratory Function Tests; Surveys and Questionnaires; Tumor Necrosis Factor-alpha

IL-8 is a strong inductor of inflammation. Accordingly, it plays a pivotal role in acute inflammatory responses during respiratory syncytial virus (RSV) infections and in chronic inflammatory diseases such as bronchial asthma and juvenile idiopathic arthritis. Recently, 2 studies have found association of the polymorphism -251A of IL8 with RSV bronchiolitis. Furthermore, epidemiologic studies have demonstrated an increased risk for the development of asthma after RSV bronchiolitis, and a common genetic background for the 2 diseases is currently being discussed.. This study investigated whether IL-8 is in association with asthma and/or arthritis and whether the results can confirm a common genetic background of RSV bronchiolitis and asthma.. The polymorphisms -A251T, C781T, C1633T, and A2767T within IL8 were genotyped in the following 4 populations: children with asthma, atopic children, children with juvenile idiopathic arthritis, and control subjects. Statistical analysis made use of the Armitage trend test and the software program Arlequine.. Association of all polymorphisms was found with asthma ( P =.008 to P =.03). Surprisingly -251T was associated with asthma, which is the opposite allele as described in association with RSV bronchiolitis. Furthermore, all polymorphisms were significantly more common in children with arthritis than in asthmatic children ( P =.006 to P =.02). No association was seen with the diagnosis of arthritis per se or with atopy.. This is the first study to describe association of IL-8 with asthma and a significant inverse distribution of the polymorphisms in juvenile idiopathic arthritis. In addition, the results of this study might suggest that RSV bronchiolitis and bronchial asthma have at least some different genetic factors.

Topics: Adolescent; Adult; Arthritis, Juvenile; Asthma; Bronchiolitis, Viral; Child; Child, Preschool; Genetic Predisposition to Disease; Genotype; Haplotypes; Humans; Interleukin-8; Polymorphism, Genetic; Respiratory Syncytial Virus Infections; Respiratory Syncytial Virus, Human

Although IL-18 was initially regarded as a factor that enhances IFN-gamma production from Th1 cells, later studies revealed its potential to induce Th2 cytokine production from T cells, NK cells and basophils/mast cells. Very recently, we demonstrated that passively transferred memory phenotype Th1 cells induce airway inflammation and hyperresponsiveness in a host mouse by production of Th1-, Th2-cytokines, GM-CSF and chemokines, when the transferred cells are stimulated in the host mice with nasally administered Ag and IL-18. Moreover, IL-18 is suggested to contribute to asthma exacerbation in human patients. Therefore, it is important to determine whether human Th1 cells also have the potential to produce these soluble factors when stimulated with anti-CD3 and IL-18 in vitro. Here we demonstrated that only Th1 cells, but not Th2 cells, produce IFN-gamma, IL-13, GM-CSF and IL-8 after stimulation with anti-CD3 and IL-18. Furthermore, highly purified IFN-gamma-producing Th1 cells have the same potential. Thus, human Th1 cells may become very harmful cells, when stimulated with Ag and IL-18 in vivo, and produce IFN-gamma, IL-13, GM-CSF and IL-8, which in combination might induce severe inflammation such as airway inflammation.

Topics: Antibodies; Asthma; CD3 Complex; Cytokines; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interferon-gamma; Interleukin-18; Interleukin-8; Th1 Cells; Th2 Cells

The role of leukotrienes (LTs) in the pathophysiology of isocyanate-induced asthma is not well known.. We sought to characterize the type of airway inflammation induced by exposure to isocyanates and to investigate whether exposure to isocyanates induced an increase in LT receptor cysteinyl leukotriene ((CysLT)(1), CysLT(2) and leukotriene B(4) receptor (BLT(1))) expression, as well as a release of LT (LTC(4) and leukotriene B(4) (LTB(4))) and IL-8 in both asthmatics with isocyanate-induced asthma and healthy subjects.. We investigated eight subjects with isocyanate-induced asthma and eight healthy subjects. Both groups underwent specific inhalation challenges to isocyanates in the laboratory. Induced sputum was collected before and after exposure to isocyanates. CysLT(1), CysLT(2) and BLT(1) expression was assessed by flow cytometry, whereas LTC(4), LTB(4) and IL-8 were measured in the sputum supernatants by enzyme immunoassay.. Exposure to isocyanates induced an increase in sputum neutrophils only in subjects with occupational asthma. There was a significant increase in CysLT(1) and BLT(1) receptor expression, as well as a release of LTB(4) and IL-8 after exposure to isocyanates compared with the baseline, only in subjects with isocyanate-induced asthma, whereas there was no increase in LTC(4). Exposure to isocyanates did not induce any change in LT receptor expression nor in the levels of LTC(4), LTB(4) and IL-8, in healthy subjects.. The neutrophilia observed after exposure to isocyanates is likely to be related to the release of LTB(4), probably enhanced by the increased expression of BLT(1) on neutrophils as well as by the release of IL-8. The significance of the increase of CysLT1 receptor expression on neutrophils is unknown and needs further investigation.

Topics: Adult; Asthma; Bronchial Provocation Tests; Cysteine; Female; Humans; Interleukin-8; Isocyanates; Leukotrienes; Macrophages; Male; Middle Aged; Neutrophils; Occupational Diseases; Pilot Projects; Receptors, Leukotriene B4; Receptors, Purinergic P2; Sputum

The study aims to investigate the changes of interleukin (IL)-17 in induced sputum, and to observe the correlation between concentrations of IL-17 and the number of inflammatory cells in induced sputum in chronic obstructive pulmonary disease (COPD) and in asthma.. Induced sputum was obtained in patients with COPD both during acute exacerbation and stable stage and in asthma during acute attack. Healthy nonsmoking volunteers were included as controls. The concentrations of IL-17 in induced sputum were measured by enzyme-linked immunosorbent assay.. The concentrations of IL-17 both in patients with COPD during acute exacerbation and with asthma were significantly higher than that in the control subjects (P < 0.001). The levels of IL-17 in patients with COPD during acute exacerbation positively correlated with that of IL-8 (r = 0.381, P = 0.038) and with the percentage of neutrophils (r = 0.446, P = 0.010) respectively. There was also a positive correlation between the concentrations of IL-17 and the numbers of eosinophils in patients with asthma.. The concentrations of IL-17 in patients with acute exacerbation of COPD and in patients with asthma were significantly increased. IL-17 may play a role in the airway inflammation in both COPD and asthma.

Topics: Adult; Aged; Asthma; Female; Humans; Interleukin-17; Interleukin-6; Interleukin-8; Leukocyte Count; Lung Diseases, Obstructive; Male; Middle Aged; Sputum

Asthma is a chronic inflammatory disease of the airways in which many cell types play a role. Although the most important cells are eosinophils, there are suggestions that also neutrophils may play a role in asthma. The aim of the study was to measure and compare chemotactic activity of neutrophils in patients with severe asthma and with COPD. We examined 49 patients with severe asthma and 23 patients with COPD. The mean number of neutrophils in peripheral blood of 20 asthmatics with irreversible airflow obstruction was 3.96 x 10(6)/ml. The chemotactic activity of neutrophils to FMLP was 2.69 SEM +/- 0.4, to IL-8 in concentration 10-7 microg/ml - 1.64, SEM +/- 0.2, and to IL-8 in concentration 10-8 microg/ml - 1.17, SEM +/- 0.1. The mean number of neutrophils in 29 asthmatics with reversible airflow obstruction was 3.08 x 10(6)/ml. Their chemotactic activity to fMLP was 1.7, SEM +/- 0.1 to IL-8 in concentration 10-7 microg/ml - 1.51. SEM +/- 0.2, and to IL-8 in concentration 10-8 microg/ml - 1.08, SEM +/- 0.1. The mean number of neutrophils in COPD patients was 4.05 x 10(6)/ml and their chemotactic activity to FMLP was 1.9, SEM +/- 0.1 to IL-8 - 1.35, SEM +/- 0.1. All asthmatic patients were treated with inhaled corticosteroids and some of them with oral corticosteroids. Despite of that treatment the number of neutrophils isolated from patients with asthma with irreversible airflow obstruction was almost the same like in COPD patients and chemotactic activity of neutrophils in this group was the highest. We concluded that corticosteroids treatment did not diminished chemotactic activity of neutrophils isolated from patient suffering from asthma with irreversible airflow obstruction.

Topics: Adolescent; Adult; Asthma; Chemotaxis, Leukocyte; Child; Child, Preschool; Female; Humans; Interleukin-8; Male; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Pulmonary Disease, Chronic Obstructive; Severity of Illness Index

To evaluate the relationship between inflammatory markers and severity of asthma in children, the amount of interleukin-8 (IL-8) and granulocyte/macrophage colony-stimulating factor (GM-CSF) released by peripheral blood mononuclear cells, exhaled nitric oxide (FE NO) levels, p65 nuclear factor-kappaB subunit, and phosphorylated IkBalpha expression by peripheral blood mononuclear cells were assessed in six control subjects, 12 steroid-naives subjects with intermittent asthma, and 17 children with moderate asthma. To investigate their predictive value, biomarker levels were correlated with the number of exacerbations during a 18-month follow-up period. We found that GM-CSF release was higher in moderate and intermittent asthmatics than in control subjects, whereas IL-8 release was higher in moderate than in intermittent asthmatics and control subjects. FE NO levels were similar among study groups. In moderate asthmatics, IL-8, GM-CSF, and FE NO significantly correlated with the exacerbation numbers. Moreover, p65 and phosphorylated IkBalpha levels were greater in moderate than in intermittent asthmatics and control subjects. According to GM-CSF, IL-8, and FE NO levels, two distinct subgroups of moderate asthmatics (low and high producers) were identified. High producers experienced more exacerbations than low producers. This study shows ongoing inflammation associated with biological and clinical heterogeneity in moderate asthmatics despite regular treatment and proposes that large prospective studies confirm the importance of biomarkers to assess inflammation and asthma control in children with asthma.

Topics: Adolescent; Albuterol; Androstadienes; Anti-Inflammatory Agents; Asthma; Biomarkers; Bronchodilator Agents; Calcium-Binding Proteins; Child; Female; Fluticasone; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Leukocytes, Mononuclear; Male; Membrane Glycoproteins; Nerve Tissue Proteins; NF-kappa B; Nitric Oxide; Receptors, Cell Surface; Salmeterol Xinafoate; Synaptotagmin I; Synaptotagmins

The extent of epithelial injury in asthma is reflected by expression of the epidermal growth factor receptor (EGFR), which is increased in proportion to disease severity and is corticosteroid refractory. Although the EGFR is involved in epithelial growth and differentiation, it is unknown whether it also contributes to the inflammatory response in asthma.. Because severe asthma is characterized by neutrophilic inflammation, we investigated the relationship between EGFR activation and production of IL-8 and macrophage inhibitory protein-1 alpha (MIP-1alpha) using in vitro culture models and examined the association between epithelial expression of IL-8 and EGFR in bronchial biopsies from asthmatic subjects.. H292 or primary bronchial epithelial cells were exposed to EGF or H2O2 to achieve ligand-dependent and ligand-independent EGFR activation; IL-8 mRNA was measured by real-time PCR and IL-8 and MIP-1alpha protein measured by enzyme-linked immunosorbent assay (ELISA). Epithelial IL-8 and EGFR expression in bronchial biopsies from asthmatic subjects was examined by immunohistochemistry and quantified by image analysis.. Using H292 cells, EGF and H2O2 increased IL-8 gene expression and release and this was completely suppressed by the EGFR-selective tyrosine kinase inhibitor, AG1478, but only partially by dexamethasone. MIP-1alpha release was not stimulated by EGF, whereas H2O2 caused a 1.8-fold increase and this was insensitive to AG1478. EGF also significantly stimulated IL-8 release from asthmatic or normal primary epithelial cell cultures established from bronchial brushings. In bronchial biopsies, epithelial IL-8, MIP-1alpha, EGFR and submucosal neutrophils were all significantly increased in severe compared to mild disease and there was a strong correlation between EGFR and IL-8 expression (r = 0.70, P < 0.001).. These results suggest that in severe asthma, epithelial damage has the potential to contribute to neutrophilic inflammation through enhanced production of IL-8 via EGFR- dependent mechanisms.

Topics: Adult; Asthma; Biopsy; Bronchi; Cells, Cultured; Chemokine CCL3; Chemokine CCL4; Chronic Disease; Epithelial Cells; ErbB Receptors; Female; Gene Expression; Humans; Inflammation; Interleukin-8; Macrophage Inflammatory Proteins; Male; Middle Aged; Neutrophils; Phosphorylation; RNA, Messenger

To investigate the pathogenic mechanism of late asthmatic response in comparison to early asthmatic response, changes of serum neutrophil chemotactic activity (NCA) using the Boyden chamber method and histamine level using the automated fluorometric analyzer were observed in 13 aspirin (ASA)-sensitive asthma subjects (group I: 7 early responders and group II: 6 dual responders) during lysine aspirin bronchoprovocation test (L-ASA BPT). Sera were collected before, and 30 min and 240 min after L-ASA BPT. Serum NCA increased significantly after 30 min (p=0.02) and decreased significantly at 240 min (p=0.02) in group I, while serum NCA of group II increased significantly at 30 min (p=0.04), tending to increase further up to 240 min with no statistical significance. NCA at 240 min in group II subjects was significantly higher than baseline NCA (p=0.02). The serum NCAs collected before and 240 min were significantly higher in group II than in group I (p<0.05, respectively). There were no significant changes in serum histamine levels during L-ASA BPT in both groups. NCA derived from mast cell may contribute to the development of early asthmatic response induced by L-ASA inhalation. There may be a possible involvement of NCA derived from mononuclear cells during late asthmatic response.

Topics: Adult; Aged; Aspirin; Asthma; Bronchial Provocation Tests; Chemotactic Factors; Chemotaxis; Female; Histamine; Humans; Interleukin-8; Lysine; Male; Mast Cells; Methacholine Chloride; Middle Aged; Monocytes; Neutrophils; Time Factors

Airway inflammation is a central feature of asthma and chronic obstructive pulmonary disease. Reactive oxygen species (ROS) contribute to inflammation by damaging DNA, which, in turn, results in the activation of poly(ADP-ribose) polymerase-1 (PARP-1) and depletion of its substrate, nicotinamide adenine dinucleotide. Here we show that prevention of PARP-1 activation protects against both ROS-induced airway epithelial cell injury in vitro and airway inflammation in vivo. H(2)O(2) induced the generation of ROS, PARP-1 activation and concomitant nicotinamide adenine dinucleotide depletion, and release of lactate dehydrogenase in A549 human airway epithelial cells. These effects were blocked by the PARP-1 inhibitor 3-aminobenzamide (3-AB). Furthermore, 3-AB inhibited both activation of the proinflammatory transcription factor nuclear factor-kappaB and expression of the interleukin-8 gene induced by H(2)O(2) in these cells. In a murine model of allergen-induced asthma, 3-AB prevented airway inflammation elicited by ovalbumin. Moreover, PARP-1 knockout mice were resistant to such ovalbumin-induced inflammation. These protective effects were associated with an inhibition of expression of the inducible nitric oxide synthase. These results implicate PARP-1 activation in airway inflammation, and suggest this enzyme as a potential target for the development of new therapeutic strategies in the treatment of asthma as well as other respiratory disorders such as chronic obstructive pulmonary disease.

Topics: Allergens; Animals; Asthma; Benzamides; Cell Line; Enzyme Inhibitors; Gene Expression; Humans; Hydrogen Peroxide; Inflammation; Interleukin-8; L-Lactate Dehydrogenase; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; NF-kappa B; Nitric Oxide Synthase; Ovalbumin; Oxidants; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Transcriptional Activation

The ability of chemokines to regulate Th1 and Th2 responses suggests a role in the pathogenesis of atopic disorders such as allergic asthma where Th2 response dominance has been observed. Although the impact of allergic asthma on local chemokine production in the lung has been the subject of investigation, little is know about the influence of disease progression on peripheral chemokine production. We now report use of whole blood culture and flow cytometry to assess the influence of mild allergic asthma on peripheral T-cell chemokine expression.. Study participants included patients with mild allergic asthma (n = 7) and nonasthmatic controls (n = 7). Following in vitro stimulation of peripheral venous blood with phorbol 12-myristate acetate (PMA) and ionomycin, flow cytometry was used to estimate the percentage of CD4+ and CD8+ T cells producing a number of chemokines, including macrophage inflammatory proteins MIP-1alpha and MIP-1beta, RANTES (regulated on activation, T-cell expressed and secreted), monocytic chemotactic protein-1 (MCP)-1, and interleukin (IL)-8, or the cytokines interferon (IFN)-gamma and IL-4. Serum levels of MIP-1alpha, MIP-1beta, RANTES, MCP-1, IL-8, IFN-gamma and IL-4 were also assessed by quantitative ELISA.. Intracellular expression of MIP-1beta by CD4+ and CD8+ T cells from allergic asthmatics was significantly reduced in comparison to that observed for nonasthmatics (median = 2.29% (1.75-3.50) vs 4.57% (3.38-6.64), P = 0.05; 14.20% (13.18-17.88) vs 44.10% (30.38-48.70), P = 0.01). Similarly, intracellular expression of MIP-1alpha by CD8+ T cells from allergic asthmatics was also significantly lower (3.67% (1.17-5.42) vs 17.10% (4.97-20.43), P = 0.05). Conversely, IL-8 expression by both CD4+ and CD8+ T cells from allergic asthmatics demonstrated significant enhancement (9.93% (7.77-11.28) vs 4.14% (3.61-7.11), P = 0.05; 8.40% (6.97-10.04) vs 4.98% (3.37-6.08), P = 0.05). Examination of intracellular IFN-gamma and IL-4 revealed no significant difference in the expression of either cytokine by CD4+ T-cells from allergic asthmatics and nonasthmatics. In contrast, expression of IFN-gamma was significantly reduced in CD8+ T-cells from allergic asthmatics (24.60% (21.08-32.50) vs 48.40% (41.50-55.28), P = 0.01).. The occurrence in mild allergic asthma of peripheral T-cell chemokine expression suggestive of a diminished Th1 response, coinciding with marginal change in cytokine profiles indicative of a Th2 response bias, confirms the importance of chemokine involvement in the etiology of allergic asthma. The ability to use whole blood culture to estimate chemokine expression in T cell subsets may ultimately provide a practical means to evaluate disease status and to monitor early intervention therapies which target chemokines.

Topics: Adult; Asthma; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Chemokine CCL3; Chemokine CCL4; Female; Humans; Interleukin-8; Macrophage Inflammatory Proteins; Male

The T helper type-2 (Th2)-dominated situation can be observed in allergic diseases such as asthma or atopic dermatitis. A reduced ability to produce IL-12, which is a key cytokine for the induction of Th1 responses, has been proposed to lead to aberrant Th2 development in these disease conditions.. This study was intended to examine how IL-12-producing ability might associate with allergic diseases as a function of age.. IL-12 production by monocytes at various ages was assessed in patients with bronchial asthma and/or atopic dermatitis (n = 100) in comparison with non-allergic control subjects (n = 144). Whole blood cells were stimulated with lipopolysaccharide (LPS) after priming with IFN-gamma, then intracellular cytokine expression of IL-12 and IL-8 as a control cytokine of CD14-positive cells was assessed by flow cytometric analysis.. In the control subjects, the ability of monocytes to produce IL-12 was negligible at birth and gradually increased with advancing age, whereas IL-8 production was intense throughout the human life. At more than 7 years of age, IL-12 production of patients with allergic diseases was significantly lower compared with that of control subjects. The unexpected finding was that infants and children below 6 years of age with allergic diseases tended to produce more IL-12 compared with age-matched controls. In this young group, it was noted that enhanced IL-12 production by monocytes was especially observed in allergic patients with specific IgE antibodies against some food allergens. Significant inverse relationships between serum IgE levels and IL-12-producing ability were found in the teenage and adult groups, but not in the younger children.. IL-12 appeared to play different roles in the pathogenesis of allergic diseases between younger and older ages.

Topics: Adolescent; Adult; Aging; Asthma; Child; Child, Preschool; Dermatitis, Atopic; Female; Flow Cytometry; Humans; Hypersensitivity; Infant; Infant, Newborn; Interferon-gamma; Interleukin-12; Interleukin-8; Lipopolysaccharide Receptors; Lipopolysaccharides; Male; T-Lymphocytes; Th1 Cells; Th2 Cells

Chemokines and T-lymphocytes play an important role in lower respiratory tract inflammation. This study evaluated the concentration of IL-8 and count of T-lymphocytes expressing adhesion molecules LFA-1, Mac-1, Lsel and their correlation in patients with asthma and COPD in periods of exacerbation and clinical improvement (after seven days of anti-inflammatory treatment). In all subjects bronchoscopic examination with BAL procedure were done in exacerbation period and after seven days of treatment. The concentration of IL-8 was measured by ELISA, and the expression of adhesion molecules by biotin-streptavidin methods. The highest concentration of IL-8 was observed in asthma patients in clinical improvement period, and the highest count of T-lymphocytes was observed in patients with COPD in remission phase. Increased concentration of chemokines could have been influenced by type of treatment administered, especially beta 2-mimetics. The significant correlation observed in COPD patients between IL-8 concentration and counts of T-cells expressing LFA-1 (r = 0.44), Mac-1 (r = 0.49), Lsel (r = 0.42) in exacerbation period suggest a chemotactic influence of IL-8 on T-lymphocytes.

Topics: Acute Disease; Adult; Asthma; Bronchoalveolar Lavage Fluid; Case-Control Studies; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; L-Selectin; Lymphocyte Function-Associated Antigen-1; Macrophage-1 Antigen; Male; Middle Aged; Pulmonary Disease, Chronic Obstructive; T-Lymphocytes; Time Factors

Epithelial-neutrophil activating peptide-78 (ENA-78) induces neutrophil migration, an early response to viral infection. Rhinovirus serotype 16 (RV16) was used to infect primary bronchial epithelial cells and a cell line (BEAS-2B). Release of ENA-78 protein was measured by enzyme-linked immunosorbent assay, ENA-78 mRNA production was quantified by reverse-transcription polymerase chain reaction, and ENA-78 promoter activity was assessed by use of a promoter construct. After infection with RV16, ENA-78 protein and mRNA increased significantly, and RV16 induced 3-fold increases in ENA-78 gene transcription. Nasal ENA-78 measured in patients with asthma with and without RV infection was more elevated in patients with RV infection present. Our study demonstrates that ENA-78 is produced in bronchial epithelial cells in response to RV16 infection. With other chemokines, it may be an important initiator of neutrophil airway inflammation during RV common colds and thus may play a role in the development of virus-associated airway pathologies.

Topics: Adult; Asthma; Bronchi; Cell Line; Cells, Cultured; Chemokine CXCL5; Chemokines, CXC; Epithelium; Female; Gene Expression Regulation; Humans; Interleukin-8; Male; Picornaviridae Infections; Promoter Regions, Genetic; Rhinovirus; RNA, Messenger; Time Factors; Transcription, Genetic; Virus Replication

Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity reaction to the fungus Aspergillus fumigatus, causing severe asthma that may progress to bronchiectasis. Sputum neutrophilia can occur in association with sputum eosinophilia and correlates with the degree of bronchiectasis. The mechanisms of sputum neutrophilia in ABPA are not known. The aim of this study was to investigate the role of the chemokine interleukin (IL)-8 in sputum neutrophilia in ABPA. Induced sputum was obtained from subjects with ABPA (n=29), and compared to nonsensitised asthma (n=9) and healthy controls (n=21). Semiquantitative polymerase chain reaction was used to assess IL-8 gene expression in induced sputum and IL-8 protein was measured by enzyme-linked immunosorbent assay. Sputum IL-8 protein was significantly higher in ABPA compared to asthma and controls. IL-8 messenger ribonucleic acid/glyceraldehyde-3-phosphate dehydrogenase ratio was elevated in ABPA compared to asthma and controls. Sputum IL-8 correlated with sputum neutrophils, matrix metalloproteinase-9 levels and forced expiratory volume in one second. Interleukin-8 gene expression and protein release were increased in allergic bronchopulmonary aspergillosis and correlated with airway neutrophilia and airway obstruction. The interleukin-8-mediated neutrophil influx in allergic bronchopulmonary aspergillosis may induce lung damage via release of matrix metalloproteinase-9, potentially leading to bronchiectasis.

Topics: Adult; Analysis of Variance; Aspergillosis, Allergic Bronchopulmonary; Aspergillus fumigatus; Asthma; Case-Control Studies; Chi-Square Distribution; Enzyme-Linked Immunosorbent Assay; Eosinophils; Female; Gene Expression; Humans; Interleukin-8; Male; Matrix Metalloproteinase 9; Middle Aged; Neutrophils; Polymerase Chain Reaction; Respiratory Function Tests; Sputum; Tomography, X-Ray Computed

Asthma is characterized by an increased production of eosinophil-active C-C chemokines by the airway epithelium. Recent studies have identified the presence of important quantities of labile zinc in the conducting airways. We hypothesized that modulation of this labile zinc could influence the production of proinflammatory chemokines in respiratory epithelial cells. The zinc chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) and the heavy metal chelator 2,3-dimercapto-1-propanesulfonic acid (DMPS) were used to reduce the labile zinc content of A549, BEAS-2B, and HFL-1 cells. Northern blot analysis and RNase protection assay were used to study the effects of the zinc chelators on mRNA expression. DMPS and TPEN specifically inhibited the production of eotaxin, regulated on activation, normal T-cell expressed, and presumably secreted, and monocyte chemotactic protein-1 in TNF-alpha-stimulated respiratory epithelial cells and fibroblasts through labile zinc chelation. The inhibitory effects of DMPS and TPEN were associated with the decreased binding of the zinc-finger transcription factor GATA-1, whereas no change in NF-kappaB activation was observed. Together these results demonstrate that modulation of the labile pool of zinc can regulate gene expression and protein synthesis of C-C chemokines in lung epithelial cells and fibroblasts.

Topics: Asthma; Chelating Agents; Chemokine CCL11; Chemokine CCL2; Chemokine CCL5; Chemokines, CC; DNA-Binding Proteins; Erythroid-Specific DNA-Binding Factors; Ethylenediamines; Fibroblasts; GATA1 Transcription Factor; Gene Expression; Humans; Interleukin-8; NF-kappa B; Protein Binding; Respiratory Mucosa; RNA, Messenger; Transcription Factors; Tumor Cells, Cultured; Unithiol; Zinc

The transcription factor nuclear factor-kappaB (NF-kappaB) is inactive when bound to its inhibitory protein IkappaBalpha. On cell stimulation with inflammatory signals, IkappaBalpha is phosphorylated by IkappaB kinases and subsequently degraded. Freed NF-kappaB then induces expression of cytokines such as granulocyte-macrophage colony-stimulating factor, interleukin-8, and regulated upon activation, normal T cell expressed and secreted. These mediators are overexpressed in asthma and are downregulated by glucocorticoids through NF-kappaB activity repression. However, high levels of granulocyte-macrophage colony-stimulating factor, interleukin-8, and regulated upon activation, normal T cell expressed and presumably secreted are released by peripheral blood mononuclear cells isolated from patients with severe asthma despite continuous systemic glucocorticoid treatment. We report that these mediators are markedly decreased by pyrrolidinedithiocarbamate, an inhibitor of NF-kappaB activation. To further characterize the persistent NF-kappaB activation in severe asthma, we analyzed the expression of various components of this activation pathway in healthy subjects and in asthmatics with mild controlled, and moderate and severe uncontrolled disease. We found high amounts of phosphorylated IkappaBalpha characterizing the three asthmatic groups. Western blot analyses indicated that in peripheral blood mononuclear cells the IkappaB kinase beta and p65 levels were greater in moderate and severe asthmatics than in normal subjects. Electrophoretic mobility shift assay and immunocytochemistry showed a greater activation status of p65 in severe asthmatics. Our data suggest that exaggerated NF-kappaB activation perpetuates inflammatory mediators production in severe asthma.

Topics: Adult; Asthma; Chemokine CCL5; Female; Glucocorticoids; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; In Vitro Techniques; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; NF-kappa B; Prednisolone; Pyrrolidines; Severity of Illness Index; Signal Transduction; Thiocarbamates

To study the clinical features and airway inflammation in eosinophilic bronchitis (EB) and the treatment outcomes.. Irwin's anatomic protocol for diagnosing chronic cough was used in 86 patients with chronic cough, and induced sputum by hypertonic saline aerosol inhalation was performed. Differential cell counts were performed in induced sputum, and eosinophilic cationic protein (ECP) was measured with fluoroimmunoassay, while interleukin-8 (IL-8) was measured with enzyme-linked absorbed immunoassay. EB was diagnosed according to Gibson's criteria and treated with inhaled budesonide 200 - 400 micro g twice daily for four weeks, and in some patients oral prednisone 10 - 15 mg/d or methyl-prednisone 8 - 12 mg/d was given for one week.. 13 (15%) out of 86 patients with chronic cough were diagnosed as having EB. Dry cough was the major compliant and all had normal lung function with negative histamine provocation test. The Eos count was 0.1862 +/- 0.1632 and the concentration of ECP (2.53 +/- 2.07) mg/L in induced sputum were significant higher in patients with EB as compared with those normal subjects (P < 0.01). The cough disappeared in all patients at the end of one week of inhaled or orally administered corticosteroids.. EB, an eosinophilic airway inflammation, is one of important causes of chronic cough and responds well to corticosteroid therapy.

Topics: Adolescent; Adult; Aged; Asthma; Blood Proteins; Bronchitis; Cough; Eosinophil Granule Proteins; Eosinophilia; Female; Humans; Interleukin-8; Male; Middle Aged; Ribonucleases

This investigation was designed to confirm IL-8 production from human bronchial epithelial cells with toluene diisocyanate (TDI) exposure and to examine the effects of pro-inflammatory cytokine and dexamethasone. We cultured Beas-2B, a bronchial epithelial cell line with TDI-HSA conjugate and compared with those without conjugate. IL-8 in the supernatant was measured by ELISA. To evaluate the effect of proinflammatory cytokines, peripheral blood mononuclear cells (PBMC) were collected from TDI- and non-TDI asthma patients, and were added to the epithelial cell culture. Dexamethasone or antibodies to TNF-alpha and IL-1beta were pre-incubated with PBMC supernatant. There was a significant production of IL-8 from bronchial epithelial cells with addition of TDI-HSA conjugate in a dose-dependent manner, which was significantly augmented with addition of PBMC supernatant. Higher production of IL-8 was noted with addition of PBMC supernatant from TDI-asthma patients than in those from non-TDI asthma patients. IL-1beta and IL-1beta/TNF alpha antibodies were able to suppress the IL-8 productions. Pre-treatment of dexamethasone induced dose-dependent inhibition of the IL-8 production. These results suggest that the IL-8 production from bronchial epithelial cells contribute to neutrophil recruitment occurring in TDI-induced airway inflammation. IL-1beta released from PBMC of TDI-induced asthma patients may be one of the pro-inflammatory cytokines to enhance IL-8 production.

Topics: Asthma; Bronchi; Cell Line; Dexamethasone; Epithelial Cells; Glucocorticoids; Humans; Interleukin-8; Leukocytes, Mononuclear; Toluene 2,4-Diisocyanate

Interleukin-8 (IL-8), a potent chemotactic and activating factor for neutrophils and eosinophils, may be a crucial factor in severe asthma. The aim of this study was to evaluate the effect of fluticasone propionate (FP), a corticosteroid with potent anti-inflammatory activity, on the in vitro release of IL-8 by monocytes obtained from asthmatic patients.. Monocytes from 15 non-atopic healthy donors and from 15 patients affected by moderate-severe allergic asthma were isolated and incubated (37 degrees C, 5% CO(2)) for 24 h with varying combinations of lipopolysaccharide (LPS) from Escherichia coli (1 microg/ml) and FP (100 nmol/l). IL-8 concentration in the culture supernatant was measured by an immuno-enzymatic method (ELISA).. A highly significant inverse correlation between FEV1 (forced expiratory volume) values before withdrawal and in vitro IL-8 production by unstimulated monocytes from asthmatic patients was observed (Rho = -0.787; p = 0.0032). IL-8 production by either LPS-stimulated or unstimulated monocytes from asthmatic subjects was statistically increased compared to monocytes from healthy donors (p < 0.05). FP addition reduced IL-8 production by monocytes from asthmatic patients and also from healthy donors (p < 0.05).. The partial IL-8 inhibition by FP could be closely related to the anti-inflammatory activity of this corticosteroid. Based on these results, we propose that the clinical improvement of asthma, observed following FP administration, may depend, at least in part, on the ability of this drug to modulate cytokine production by monocytes.

Topics: Adolescent; Adult; Analysis of Variance; Androstadienes; Anti-Allergic Agents; Anti-Inflammatory Agents; Asthma; Cells, Cultured; Female; Fluticasone; Humans; Interleukin-8; Male; Middle Aged; Monocytes; Statistics, Nonparametric

Chronic diseases may involve an "innate" response followed by an adaptive immune response, of a Th1 or Th2 variety. Little is known regarding the interactions of these responses. We hypothesized that TGF-beta1 (innate response factor associated with wound repair) in combination with IL-13 (Th2 factor) might augment inflammatory processes associated with asthma. Airway fibroblasts were cultured from asthmatic subjects and normal controls. These fibroblasts were exposed to TGF-beta1 and IL-13 alone or in combination, and eotaxin-1 expression and production were evaluated. At 48 h, eotaxin-1 production was markedly increased with the combination of TGF-beta1 and IL-13 (p < 0.0001) compared with either stimulus alone. mRNA increased slightly at 1 h with IL-13 or TGF-beta1 plus IL13, peaked, and became significantly increased over IL-13 alone at 24 h. Protein was measurable from 6 h with IL-13 and TGF-beta1 plus IL-13, but greater levels were measured over time with the combination. Actinomycin ablated the increase in mRNA and protein seen with IL-13 alone and with TGF-beta1 plus IL-13. Cycloheximide blocked the increase in mRNA at 6 h in both conditions, but also blocked the increase at 24 h with TGF-beta1 plus IL-13. STAT-6 was rapidly activated with both IL-13 and the combination, without difference. Finally, eotaxin-1-positive fibroblasts were identified in severe asthma biopsies in greater numbers than in normals. These results support the concept that interactions of innate and adaptive immune systems may be important in promoting the tissue eosinophilia of asthma, particularly in those with more severe disease.

Topics: Adjuvants, Immunologic; Asthma; Blotting, Northern; Bronchi; Cells, Cultured; Chemokine CCL11; Chemokines, CC; Chemotactic Factors, Eosinophil; Cycloheximide; Dactinomycin; Dose-Response Relationship, Immunologic; Drug Synergism; Fibroblasts; Humans; Interleukin-13; Interleukin-8; Lung; Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; STAT6 Transcription Factor; Trans-Activators; Transforming Growth Factor beta; Transforming Growth Factor beta1; Up-Regulation

Air pollution constitutes an important factor for asthma aggravation, and there is increased concern about respiratory health effects of common air pollutants. The purpose of this study was to examine how exposure to a high ambient concentration nitrogen dioxide (NO2) prior to a bronchial allergen challenge modulated the inflammatory response in the bronchi. Thirteen subjects with mild asthma and allergy were exposed at rest to either purified air or 500 microg x m 3 NO2 for 30 min, followed 4 h later by an allergen inhalation challenge. The exposures (NO2 or air) were performed in random order and at least 4 weeks apart. Lung function during NO2/air exposure and allergen challenge was measured by plethysmography, and then hourly by portable spirometry after exposures. Subjective symptoms were recorded during and after exposure. Bronchoscopy with bronchial wash (BW) and bronchoalveolar lavage (BAL) was performed 19 h after allergen challenge. NO2+allergen enhanced the percentage of neutrophils in both BW and BAL compared to air+allergen (BW 19 vs. 11, P=0.05; BAL 3 vs. 1, P=0.02 median values). The levels of eosinophil cationic protein (ECP) in BW was higher after NO2+allergen compared to air+allergen (90 vs. 3.6 microg/l; P=0.02, median values). There was no NO2-associated effect on symptoms or pulmonary function. These data suggest that ambient NO2 can enhance allergic inflammatory reaction in the airways without causing symptoms or pulmonary dysfunction.

Topics: Adult; Airway Resistance; Albumins; Allergens; Asthma; Bronchi; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Cell Count; Eosinophils; Female; Forced Expiratory Volume; Humans; Interleukin-5; Interleukin-8; Male; Nitrogen Dioxide

The differences between respiratory syncytial virus (RSV) and influenza A virus (IFAV) in the pathogenesis of wheezing in young children have not been clearly defined. The aim of this study was to assess the contributions of RSV vs IFAV in the pathogenesis of upper airway inflammation in wheezy young children. We compared interleukin (IL)-6, IL-8, IL-11, and interferon-gamma (IFN-gamma) levels in nasopharyngeal aspirates (NPA) from non-asthmatic children with respiratory virus infections (RSV in 17 children and IFAV in 13 children), asthmatic children with viral infections (RSV in nine children, IFAV in 10 children), and 22 unaffected healthy children (controls). Levels of IL-11 in NPA from asthmatic children were significantly higher than those from non-asthmatic children with RSV infection, and RSV infection enhanced the IL-11 production in NPA significantly compared to IFAV infection. Nasopharyngeal epithelium from children with RSV infection secreted more IL-6 than that of children with IFAV infection. There was little difference in the IL-8 and IFN-gamma levels between asthmatic and non-asthmatic children with RSV or IFAV infection. In conclusion, asthma enhanced IL-11 production in RSV infection rather than IFAV infection in early childhood. There was a trend towards greater IL-6 production in RSV infection compared with IFAV infection.

Topics: Asthma; Biomarkers; Bronchiolitis, Viral; Child Welfare; Child, Preschool; Cytokines; Female; Humans; Infant; Infant Welfare; Influenza A virus; Influenza, Human; Inhalation; Interferon-gamma; Interleukin-11; Interleukin-6; Interleukin-8; Male; Nasopharynx; Respiratory Sounds; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses; Statistics as Topic

Different forms of chronic airway inflammation may involve diverse pathogenic elements. In general, deficient defence response is a feature of chronic obstructive pulmonary disease (COPD), whereas distorted immunoregulatory mechanisms lead to development of asthmatic symptoms. In addition to diverse effector mechanisms, the cellular and humoral elements participating in the development of immune response may appear to be different in COPD and bronchial asthma (BA) patients.. To evaluate the immunoregulatory properties of T cells and monocytes in cultures of peripheral blood mononuclear cells (PBMC) and to determine the chosen cytokine profiles in COPD and BA patients.. The microcultures of PBMC from COPD and BA patients were assessed for the T-cell response to mitogens, saturation of interleukin (IL)-2 receptors, T-cell suppressive activity and monokine influence on lymphocyte proliferation. Concomitantly, the cytokine (IL-1beta, interleukin-1 receptor antagonist, tumour necrosis factor-alpha, IL-4, IL-6, IL-8) concentrations were determined in the serum, the broncho-alveolar lavage fluid and in the culture supernatants.. The T-lymphocyte reactions (response to phytohaemagglutinin, IL-2 receptor saturation, suppressive activity) were lower in BA patients than in COPD patients. Reversely, the immunogenic activity of monocytes (IL-1beta versus IL-1ra production) was higher in BA patients than in COPD patients. The highest values of cytokine concentrations were found in the culture supernatants. The concentrations of tumour necrosis factor-alpha, IL-4, IL-6 and IL-8 were significantly higher and the concentration of IL-1ra was lower in BA patients than in COPD patients.. The assessments of cellular immunoregulatory properties and cytokine profiles in the cultures of blood mononuclear cells may prove helpful for diagnostic and therapeutic discrimination between BA and COPD patients.

Topics: Adult; Asthma; Bronchoalveolar Lavage Fluid; Cells, Cultured; Cytokines; Diagnosis, Differential; Female; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Interleukin-4; Interleukin-6; Interleukin-8; Male; Middle Aged; Monocytes; Pulmonary Disease, Chronic Obstructive; Sialoglycoproteins; T-Lymphocytes; Tumor Necrosis Factor-alpha

Elevated levels of exhaled nitric oxide are seen in inflammatory airway diseases such as asthma, but the cellular source remains unknown. This study investigated whether human airway epithelial cells express inducible nitric oxide synthase (iNOS). Human bronchial epithelial cells stimulated with 50 ng/ml interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma express iNOS mRNA, protein and increased nitrite in the cell culture media, which was inhibited by the selective iNOS inhibitor 1400W. Cells derived from subjects with asthma produced less nitrite than cells from normal subjects (6.59 +/- 0.99 microM nitrite, n = 15 versus 3.89 +/- 0.42 microM nitrite, n = 20; P < 0.05). This was not attributed to steroid treatment of subjects with asthma because there was no difference in the amount of nitrite released from steroid-naive and steroid-treated cells (3.51 +/- 0.46 versus 4.27 +/- 0.7 microM nitrite, n = 10). Neither dexamethasone nor budesonide inhibited iNOS mRNA induction, protein expression, or nitrite accumulation. The cells were not steroid insensitive because steroids inhibited GM-CSF release. Therefore, although these cells express iNOS under inflammatory conditions, they do not appear to be regulated directly by glucocorticosteroids.

Topics: Administration, Topical; Adrenal Cortex Hormones; Adult; Anti-Inflammatory Agents; Asthma; Blotting, Western; Bronchoscopy; Budesonide; Cell Line; Cell Survival; Cells, Cultured; Dexamethasone; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Glucocorticoids; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interferon-gamma; Interleukin-1; Interleukin-8; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitrites; Protein Biosynthesis; Respiration; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; Steroids; Time Factors; Transcription, Genetic; Tumor Necrosis Factor-alpha

Patients with food allergy (FA) have been recently shown to develop bronchial hyperresponsiveness (BHR), despite the absence of any concomitant asthmatic manifestation. In order to explain this observation, we sought to examine the presence of a bronchial inflammation in induced sputum of nonasthmatic patients with FA.. Twelve nonasthmatic patients with FA (urticaria, digestive symptoms, anaphylaxis) were included in the study. Results were compared to these obtained from eight asthmatic patients without food allergy and eight healthy controls. Diagnosis of FA was based on double-blind placebo-controlled challenge. Sputum cells and fluid-phase eosinophil cationic protein (ECP), myeloperoxidase (MPO) and interleukin-8 (IL-8) were measured in induced sputum. BHR was evaluated using methacholine inhalation.. Sputum from asthmatics, in comparison with the sputum of healthy subjects and patients with FA contained a higher proportion of eosinophils and higher levels of ECP (< 0.001). In marked contrast, patients with FA exhibited an increased proportion of neutrophils and IL-8 in comparison with asthmatics and controls (P < 0.05 for neutrophils and P < 0.001 for IL-8). There was a significant correlation between sputum neutrophils and IL-8 (r = 0.68, P < 0.001). MPO levels were not different between the groups. There was a trend toward higher levels of IL-8 and ECP in food allergic patients with BHR in comparison with patients with FA without BHR.. Our results demonstrate that a subclinical neutrophil airway inflammation is present in patients with food allergy free of clinical respiratory symptoms and that IL-8 may be an important mediator of this neutrophilia.

Topics: Adult; Asthma; Blood Proteins; Bronchial Hyperreactivity; Double-Blind Method; Eosinophil Granule Proteins; Eosinophils; Food Hypersensitivity; Humans; Immunoglobulin E; Interleukin-8; Leukocyte Count; Neutrophils; Peroxidase; Respiratory Function Tests; Ribonucleases; Skin Tests; Sputum; Statistics as Topic

Mycoplasma pneumoniae (Mp) infection is associated with asthma exacerbation in children. We hypothesized that Mp infection may cause airway inflammation by inducing the release of cytokines by respiratory epithelial cells. The levels of chemokines interleukin-8 (IL-8) and released upon activation, normal t cell expressed and secreted (RANTES) released by nasal epithelial cell (NEC) cultures established from asthmatic and nonasthmatic children were measured by ELISA at 4, 24, 48, and 72 hr after cells were inoculated with Mp, and were compared with baseline release of these factors. The presence of MP on apical membranes of NEC after infection was confirmed by transmission electron microscopy, and adherence was shown to be inhibited by erythromycin. Mp infection did not alter NEC release of IL-8 or RANTES at any time point. In contrast, tumor necrosis factor alpha (TNF-alpha) stimulated increased IL-8 at all time points, and respiratory syncytial virus (RSV) infection stimulated RANTES release at 48 and 72 hr by NEC. These results were not significantly different between NEC from asthmatic and nonasthmatic children. As a comparison, peripheral blood mononuclear cells from normal human volunteers were also incubated with Mp and had significantly increased release of IL-2, IL-6, and TNF-alpha. We conclude that Mp, unlike viral pathogens such as RSV, is unlikely to directly stimulate early airway surface cytokine responses via mechanisms involving epithelial cells. We speculate that the chronic presence of mononuclear cells at the airway surface of asthmatics provides a target for Mp-triggered cytokine production.

Topics: Asthma; Cells, Cultured; Chemokine CCL5; Child; Culture Media, Conditioned; Cytokines; Humans; Interleukin-2; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Microscopy, Electron; Mycoplasma pneumoniae; Nasal Mucosa; Pneumonia, Mycoplasma; Time Factors; Tumor Necrosis Factor-alpha

Topics: Asthma; Blood Proteins; Cell Separation; Cell Survival; Eosinophils; Humans; Interleukin-5; Interleukin-8; Peroxidase; Pulmonary Disease, Chronic Obstructive; Reproducibility of Results; Sputum

Lipoxins, endogenous eicosanoids biosynthetized in vivo at inflammation sites, are potential antiinflammatory mediators. Subjects with severe asthma present chronic inflammation of the airways despite long-term treatment with oral glucocorticoids. Therefore it is of interest to investigate the potential antiinflammatory effects of lipoxin A4 (LXA4) and lipoxin B4 (LXB4) that could attenuate chronic inflammation. In a first time, we detected interleukin (IL)-8 and LXA4 in supernatants of induced sputum. IL-8 was heightened in severe asthma (p = 0.001), whereas high concentrations of lipoxin A4 were present in mild asthma (p = 0.001). We then studied the effects of LXA4 on IL-8 released in vitro. Nanomolar concentrations of LXA4 and LXB4 inhibited the IL-8 released by peripheral blood mononuclear cells from the two groups of patients with asthma: a maximal inhibition of 29.4% (p < 0.01) was observed for patients with mild asthma, and 41.5% inhibition (p < 0.001) for patients with severe asthma at 1 nM and 100 nM LXA4 concentrations, respectively. Polymerase chain reaction analysis indicated that peripheral blood mononuclear cells from patients with asthma expressed the LXA4 receptor mRNA. Moreover, pertussis toxin reversed LXA4- and LXB4-inhibited IL-8 release. These findings suggest that lipoxins have potential antiinflammatory action in asthma.

Topics: Asthma; Base Sequence; Biomarkers; Bronchial Provocation Tests; Female; Humans; Hydroxyeicosatetraenoic Acids; Inflammation Mediators; Interleukin-8; Lipoxins; Male; Molecular Sequence Data; Probability; Prospective Studies; Reference Values; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sampling Studies; Sensitivity and Specificity; Severity of Illness Index; Sputum; Statistics, Nonparametric

In sensitized individuals, exposure to allergens such as Dermatophagoides pteronyssinus (Der p) causes Th2 polarization and release of cytokines, including IL-4 and IL-13. Because Der p extracts also have direct effects on epithelial cells, we hypothesized that allergen augments the effects of Th2 cytokines by promoting mediator release from the bronchial epithelium in allergic asthma. To test our hypothesis, primary bronchial epithelial cultures were grown from bronchial brushings of normal and atopic asthmatic subjects. RT-PCR showed that each culture expressed IL-4R(alpha), common gamma-chain, and IL-13R(alpha)(1), as well as IL-13R(alpha)(2), which negatively regulates IL-13 signaling; FACS analysis confirmed IL-13R(alpha)(2) protein expression. Exposure of epithelial cultures to either Der p extracts, TNF-alpha, IL-4, or IL-13 enhanced GM-CSF and IL-8 release, and this was partially suppressible by corticosteroids. Simultaneous exposure of the epithelial cultures to IL-4 or IL-13 together with Der p resulted in a further increase in cytokine release, which was at least additive. Release of TGF-alpha was also increased by TNF-alpha and combinations of IL-4, IL-13, and Der p; however, this stimulation was only significant in the asthma-derived cultures. These data suggest that, in an allergic environment, Th2 cytokines and allergen have the potential to sustain airway inflammation through a cooperative effect on cytokine release by the bronchial epithelium. Our novel finding that IL-4, IL-13, and allergen enhance release of TGF-alpha, a ligand for the epidermal growth factor receptor that stimulates fibroblast proliferation and goblet cell differentiation, provides a potential link between allergen exposure, Th2 cytokines, and airway remodelling in asthma.

Topics: Adult; Allergens; Animals; Antigens, Dermatophagoides; Asthma; Bronchi; Cells, Cultured; Cytokines; Drug Combinations; Female; Glycoproteins; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-13; Interleukin-13 Receptor alpha1 Subunit; Interleukin-4; Interleukin-8; Male; Middle Aged; Mites; Receptors, Interleukin; Receptors, Interleukin-13; Receptors, Interleukin-4; Respiratory Mucosa; Th2 Cells; Transforming Growth Factor alpha; Tumor Necrosis Factor-alpha

To explore the clinical value of induced sputum test in monitoring airway inflammation in asthmatic patients.. The induced sputum test was done in 12 normal subjects and 21 asthmatic subjects; The levels of interleukin-8 (IL-8), eosinophil cation protein (ECP), malondialdehyde (MDA) in plasma and induced sputum in all subjects were measured.. The levels of IL-8, ECP, MDA in plasma and induced sputum in unstable asthmatic patients were significantly higher than those in stable asthmatic (P < 0.01), in which the levels of IL-8, ECP, MDA in both plasma and induced sputum were significantly higher than those in normal subjects(P < 0.05); In all asthmatic patients, the levels of IL-8, ECP, MDA in induced sputum were significantly higher than those in plasma.. The levels of IL-8, ECP, MDA in induced sputum could be used for monitoring airway inflammation in asthmatic patients.

Topics: Adult; Asthma; Blood Proteins; Bronchi; Eosinophil Granule Proteins; Humans; Inflammation; Interleukin-8; Malondialdehyde; Ribonucleases; Sputum

Human polymorphonuclear neutrophils (PMNs) express surface receptors for various inflammatory mediators, including IgE and IL-4. Recently, the IL-9R locus has been genetically linked to asthma and bronchial hyperresponsiveness in humans. In this study, we evaluated expression of the IL-9R and the effect of IL-9 on human PMNs. RT-PCR analysis showed the presence of IL-9Ralpha-chain mRNA in PMN RNA preparations from asthmatic patients. Using FACS analysis, surface expression of IL-9Ralpha was detected on PMNs freshly isolated from asthmatics, and to a lesser extent on normal controls. In addition, protein expression of IL-9Ralpha was also detected in peripheral blood and bronchoalveolar lavage PMNs. Furthermore, functional studies showed that IL-9 stimulation of PMNs results in the release of IL-8 in a concentration-dependent manner. The anti-IL-9 neutralizing Ab suppressed this effect, but had no effect on GM-CSF-induced IL-8 release from PMNs. Taken together, these findings suggest a novel role for PMNs in allergic disease through the expression and activation of the IL-9R.

Topics: Asthma; Bronchoalveolar Lavage Fluid; Cell Membrane; HL-60 Cells; Humans; Immunohistochemistry; Interleukin-8; Interleukin-9; Neutrophils; Receptors, Interleukin; Receptors, Interleukin-9; RNA, Messenger

Although studies have suggested that ozone (O3) and nitrogen dioxide (NO2) may play a role in the pathogenesis of asthma, the underlying mechanisms are not clear.. We aimed to investigate the effects of O3 and NO2 on the release of IL-8, GM-CSF, RANTES, and soluble intercellular adhesion molecule 1 (sICAM-1) from human bronchial epithelial cells (HBECs) of nonatopic nonasthmatic subjects (nonasthmatic subjects) and atopic subjects with mild asthma (asthmatic subjects) in vitro.. We cultured HBECs from bronchial biopsy specimens of nonasthmatic and asthmatic subjects; exposed these for 6 hours to air, 10 to 100 ppb O3, or 100 to 400 ppb NO2; and analyzed the release of IL-8, GM-CSF, RANTES, and sICAM-1 after 24 hours' incubation.. There was no significant difference between the constitutive release of IL-8, GM-CSF, and sICAM-1 from HBECs of asthmatic and nonasthmatic subjects. RANTES was detected only in HBECs derived from asthmatic subjects. Exposure of HBECs of asthmatic subjects to both 50 to 100 ppb O3 and 200 to 400 ppb NO2 significantly increased the release of IL-8, GM-CSF, RANTES, and sICAM-1 from these cells after 24 hours of incubation. However, 50 to 100 ppb O3 and 200 to 400 ppb NO2 led to a significant increase in release of only IL-8 and sICAM-1 from HBECs of nonasthmatic subjects after 24 hours' incubation. A comparison between the pollutant-induced release of mediators demonstrated that 100 ppb O3-induced release of GM-CSF and sICAM-1 was significantly greater in HBECs of asthmatic subjects (medians, 0.59 and 27.4 pg/microg cellular protein, respectively) than in HBECs of nonasthmatic subjects (medians, 0.27 and 14.4 pg/microg cellular protein, respectively; P < .02).. These results suggest that O3 and NO2 may modulate airway diseases, such as asthma, by increasing the release of inflammatory mediators from bronchial epithelial cells and that the cells of asthmatic subjects may be more susceptible to the adverse effects of these pollutants.

Topics: Asthma; Bronchi; Cells, Cultured; Chemokine CCL5; Epithelial Cells; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Hypersensitivity, Immediate; Inflammation Mediators; Intercellular Adhesion Molecule-1; Interleukin-8; Nitrogen Dioxide; Ozone; Secretory Rate

It has been shown previously that airway eosinophils characterize childhood asthma and neutrophils contribute to the pathophysiology of both infantile wheezing and asthma. Therefore, eosinophil cationic protein (ECP) and interleukin-8 (IL-8) levels in bronchoalveolar lavage fluid (BALF) from asthmatics (n = 16) and infantile wheezers (n = 30) were analyzed as markers of eosinophil- and neutrophil-mediated inflammation. To aid the interpretation, a control group of children (n = 10) with no lower airway pathology were included. Disease severity was assessed by using a symptom score. Surprisingly, no significant difference was found in IL-8 or ECP levels among asthma, infantile wheeze, and control groups. Asthma was characterized by: a correlation between ECP levels and eosinophil counts (r = 0.618, p = 0.014); a correlation between neutrophil number and IL-8 levels (r = 0.747, p = 0.002); and increasing IL-8 levels with symptom score (p = 0.03). In infantile wheezers, IL-8 levels were poorly related to neutrophil number but were significantly increased when neutrophils were > 10%. Although detectable levels were found in all but one symptomatic infant, IL-8 concentrations did not reflect the symptom score in infantile wheeze. ECP was unexpectedly correlated to neutrophil percentages (Rho = 0.832, p = 0.001), and a threshold of ECP>20 ng/ml was associated with persistent symptoms in these infantile wheezers. Hence, in accordance with BALF cellularity, activation of eosinophils was suggested by raised levels of ECP in childhood asthma, but not in infantile wheeze. Neutrophil-mediated inflammation appeared to better reflect the severity of asthma than that of infantile wheeze. Although its meaning remains to be elucidated, ECP was suggested to be a helpful indicator of persistent infantile wheeze. However, its utility as a marker predicting ongoing asthma remains to be established.

Topics: Adolescent; Asthma; Biomarkers; Blood Proteins; Bronchoalveolar Lavage Fluid; Bronchoscopy; Child; Child, Preschool; Eosinophil Granule Proteins; Female; Humans; Infant; Inflammation Mediators; Interleukin-8; Male; Respiratory Sounds; Ribonucleases

Polymorphonuclear neutrophils (PMNs) are important effector cells in host defense and the inflammatory response to antigen. The involvement of PMNs in inflammation is mediated mainly by the Fc receptor family, including IgE receptors. Recently, PMNs were shown to express two IgE receptors (CD23/Fc epsilon RII and galectin-3). In allergic diseases, the dominant role of IgE has been mainly ascribed to its high-affinity receptor, Fc epsilon RI. We have examined the expression of Fc epsilon RI by PMNS: mRNA and cell surface expression of Fc epsilon RI alpha chain was identified on PMNs from asthmatic subjects. Furthermore, preincubation with human IgE Fc fragment blocks completely the binding of anti-Fc epsilon RI alpha chain (mAb15--1) to human PMNS: Conversely, preincubation of PMNs with mAb15--1 inhibits significantly the binding of IgE Fc fragment to PMNs, indicating that IgE bound to the cell surface of PMNs mainly via the Fc epsilon RI. Peripheral blood and bronchoalveolar lavage (BAL) PMNs from asthmatic subjects also express intracellular Fc epsilon RI alpha and beta chain immunoreactivity. Engagement of Fc epsilon RI induces the release of IL-8 by PMNS: Collectively, these observations provide new evidence that PMNs express the Fc epsilon RI and suggest that these cells may play a role in allergic inflammation through an IgE-dependent activation mechanism.

Topics: Asthma; HL-60 Cells; Humans; Immunoglobulin E; Immunoglobulin Fc Fragments; Immunohistochemistry; In Situ Hybridization; Interleukin-8; Leukocytes, Mononuclear; Neutrophils; Receptors, IgE; RNA, Messenger

As peripheral blood mononuclear cells from patients with nocturnal asthma (NA) exhibit reduced steroid responsiveness at 4:00 A.M. as compared with 4:00 P.M., we hypothesized that NA is associated with increased nocturnal airway cell expression of GRbeta, an endogenous inhibitor of steroid action. Ten subjects with NA and seven subjects with nonnocturnal asthma (NNA) underwent bronchoscopy with bronchoalveolar lavage (BAL) at 4:00 P.M. and 4:00 A.M. BAL lymphocytes and macrophages were incubated with dexamethasone (DEX) at 10(-5) to 10(-8) M. DEX suppressed proliferation of BAL lymphocytes similarly at 4:00 P.M. and 4:00 A.M. in both groups. However, BAL macrophages from NA exhibited less suppression of IL-8 and TNF-alpha production by DEX at 4:00 A.M. as compared with 4:00 P.M. (p = 0.0001), whereas in the NNA group DEX suppressed IL-8 and TNF-alpha production equally at both time points. GRbeta expression was increased at night only in NA, primarily due to significantly increased expression by BAL macrophages (p = 0.008). IL-13 mRNA expression was increased at night, but only in the NA group and addition of neutralizing antibodies to IL-13 reduced GRbeta expression by BAL macrophages. We conclude that the airway macrophage may be the airway inflammatory cell driving the reduction in steroid responsiveness at night in NA, and this function is modulated by IL-13.

Topics: Adult; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Circadian Rhythm; Dexamethasone; Female; Humans; Interleukin-13; Interleukin-8; Least-Squares Analysis; Lymphocytes; Macrophages; Male; Receptors, Glucocorticoid; RNA, Messenger; Tumor Necrosis Factor-alpha

To identify the characteristics of airway inflammation in persistent asthma and to examine the role of neutrophilic inflammation in noneosinophilic persistent asthma.. Nonsmoking adults (n = 56) with persistent asthma and healthy control subjects (n = 8) underwent hypertonic saline solution challenge and sputum induction. Selected sputum portions were dispersed with dithiothreitol and assayed for total cell count, cellular differential, supernatant eosinophil cationic protein (ECP), myeloperoxidase, and interleukin (IL)-8.. We identified two distinct inflammatory patterns. Typical eosinophilic inflammation occurred in 41% of subjects, whereas the remainder exhibited noneosinophilic asthma (59%). Both neutrophil percentage and absolute neutrophil counts were increased in subjects with noneosinophilic asthma (64%, 283 x 10(6)/mL) compared to eosinophilic asthma (14%, 41 x 10(6)/mL) and control subjects (34%, 49 x 10(6)/mL; p = 0.0001). Myeloperoxidase was elevated in both noneosinophilic (280 ng/mL) and eosinophilic groups (254 ng/mL) compared with control subjects (82 ng/mL; p = 0.002). Sputum IL-8 levels were highest in subjects with noneosinophilic asthma (45 ng/mL) compared to eosinophilic asthma (9.6 ng/mL) and control subjects (3.5 ng/mL; p = 0.0001). Neutrophils correlated with IL-8 levels (r = 0.72). ECP was highest in subjects with eosinophilic asthma (2,685 ng/mL) compared with noneosinophilic asthma (1,081 ng/mL) and control subjects (110 ng/mL; p = 0.0001).. Induced-sputum analysis in persistent asthma identifies two different inflammatory patterns. The most common pattern is noneosinophilic, associated with a neutrophil influx and activation, which may be mediated by IL-8 secretion. There is heterogeneity of airway inflammation in persistent asthma, which indicates differing mechanisms and may impact on treatment responses.

Topics: Adult; Asthma; Cell Count; Eosinophils; Female; Humans; Interleukin-8; Male; Middle Aged; Neutrophils; Peroxidase; Sputum

The effect of formoterol, alone and in combination with budesonide, upon tumour necrosis factor-alpha stimulated (10 ng x mL(-1)) human bronchial epithelial cells was investigated. Addition of formoterol (> or = 10(-10) M) reduced granulocyte macrophage-colony stimulating factor (GM-CSF) levels, as assessed by enzyme-linked immunosorbent assay, by 40-50% and increased interleukin (IL)-8 levels by approximately 50%. The effects of formoterol were long lasting (23 h). Budesonide (10(-8) M) reduced the amounts of both cytokines (GM-CSF and IL-8) by 40%. Simultaneous addition of formoterol and budesonide reduced GM-CSF levels approximately 75%, while IL-8 levels were decreased approximately 40%, similar to the reduction obtained with budesonide alone. The glucocorticoid receptor (GR) antagonist RU486 did not influence the effect of formoterol, suggesting no involvement of the GR. Formoterol rapidly induced an elevation in intracellular cyclic adenosine monophosphate, which was reduced in the presence of propranolol. In addition, the alterations in cytokine secretion induced by formoterol could be fully blocked by propranolol, demonstrating that these effects are beta2-receptor mediated. In conclusion, the combination of budesonide and formoterol reduces the secretion of granulocyte macrophage-colony stimulating factor to basal levels and counteracts the capacity of formoterol alone to induce interleukin-8 production, modulations which may facilitate improved asthma control.

Topics: Asthma; Bronchi; Budesonide; Cells, Cultured; Cyclic AMP; Drug Interactions; Ethanolamines; Formoterol Fumarate; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Propranolol; Respiratory Mucosa; Transforming Growth Factor alpha

The bronchial epithelium is a source of both alpha and beta chemokines and, uniquely, of secretory component (SC), the extracellular ligand-binding domain of the polymeric IgA receptor. Ig superfamily relatives of SC, such as IgG and alpha(2)-macroglobulin, bind IL-8. Therefore, we tested the hypothesis that SC binds IL-8, modifying its activity as a neutrophil chemoattractant. Primary bronchial epithelial cells were cultured under conditions to optimize SC synthesis. The chemokines IL-8, epithelial neutrophil-activating peptide-78, growth-related oncogene alpha, and RANTES were released constitutively by epithelial cells from both normal and asthmatic donors and detected in high m.w. complexes with SC. There were no qualitative differences in the production of SC-chemokine complexes by epithelial cells from normal or asthmatic donors, and in all cases this was the only form of chemokine detected. SC contains 15% N-linked carbohydrate, and complete deglycosylation with peptide N-glycosidase F abolished IL-8 binding. In micro-Boyden chamber assays, no IL-8-dependent neutrophil chemotactic responses to epithelial culture supernatants could be demonstrated. SC dose-dependently (IC(50) approximately 0.3 nM) inhibited the neutrophil chemotactic response to rIL-8 (10 nM) in micro-Boyden chamber assays and also inhibited IL-8-mediated neutrophil transendothelial migration. SC inhibited the binding of IL-8 to nonspecific binding sites on polycarbonate filters and endothelial cell monolayers, and therefore the formation of haptotactic gradients, without effects on IL-8 binding to specific receptors on neutrophils. The data indicate that in the airways IL-8 may be solubilized and inactivated by binding to SC.

Topics: Asthma; Binding Sites; Bronchi; Cells, Cultured; Chemokines; Chemotaxis, Leukocyte; Epithelial Cells; Glycosylation; Humans; In Vitro Techniques; Interleukin-8; Macromolecular Substances; Molecular Weight; Neutrophils; Secretory Component; Signal Transduction

IL-17 is a cytokine that has been reported to be produced by T lymphocytes. In vitro, IL-17 activates fibro-blasts and macrophages for the secretion of GM-CSF, TNF-alpha, IL-1beta, and IL-6. A number of these cytokines are involved in the airway remodeling that is observed within the lungs of asthmatic individuals.. In this study, we investigated the expression of IL-17 in sputum and bronchoalveolar lavage specimens obtained from asthmatic subjects and from nonasthmatic control subjects.. IL-17 was detected through use of immunocytochemistry, in situ hybridization, and Western blot. Bronchial fibroblasts were stimulated with IL-17, and cytokine production and chemokine production were detected through use of ELISA and RT-PCR.. Using immunocytochemistry, we demonstrated that the numbers of cells positive for IL-17 are significantly increased in sputum and bronchoalveolar lavage fluids of subjects with asthma in comparison with control subjects (P <.001 and P <.005, respectively). We demonstrated that in addition to T cells, eosinophils in sputum and bronchoalveolar lavage fluids expressed IL-17. Peripheral blood eosinophils were also positive for IL-17, and the level of IL-17 in eosinophils purified from peripheral blood was significantly higher in subjects with asthma than in controls (P <.01). To further investigate the mechanism of action of IL-17 in vivo, we examined the effect of this cytokine on fibroblasts isolated from bronchial biopsies of asthmatic and nonasthmatic subjects. IL-17 did enhance the production of pro-fibrotic cytokines (IL-6 and IL-11) by fibroblasts, and this was inhibited by dexamethasone. Similarly, IL-17 increased the level of other fibroblast-derived inflammatory mediators, such as the alpha-chemokines, IL-8, and growth-related oncogene-alpha.. Our results, which demonstrate for the first time that eosinophils are a potential source of IL-17 within asthmatic airways, suggest that IL-17 might have the potential to amplify inflammatory responses through the release of proinflammatory mediators such as alpha-chemokines.

Topics: Adult; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; Cytokines; Eosinophils; Female; Fibroblasts; Growth Substances; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-11; Interleukin-17; Interleukin-6; Interleukin-8; Male; Sputum

A novel gene, designated ML-1, was identified from a human genomic DNA clone and human T cell cDNA sequences. The second exon of ML-1 gene shares significant sequence identity with the gene encoding IL-17 (IL-17). ML-1 gene expression was up-regulated in activated PBMCs, CD4(+) T cells, allergen-specific Th0, Th1, and Th2 clones, activated basophils, and mast cells. Increased expression of the ML-1 gene, but not IL-17, was seen following allergen challenge in four asthmatic subjects, suggesting its role in allergic inflammatory responses. ML-1 from transiently transfected COS-7 cells was able to induce gene expression and protein production for IL-6 and IL-8 (at 10 ng/ml of ML-1: for IL-6, 599.6 +/- 19.1 pg/ml; for IL-8, 1724.2 +/- 132.9 pg/ml; and at 100 ng/ml of ML-1: for IL-6, 1005.3 +/- 55.6 pg/ml; for IL-8, 4371.4 +/- 280.5 pg/ml; p < 0.05 for both doses vs baseline) in primary bronchial epithelial (PBE) cells. Furthermore, increased expression of ICAM-1 was found in ML-1-stimulated PBE cells (mean fluorescence intensity (MFI) = 31.42 +/- 4.39 vs baseline, MFI = 12.26 +/- 1.77, p < 0.05), a functional feature distinct from IL-17 (MFI = 11.07 +/- 1.22). This effect was not inhibited by a saturating amount of IL-17. These findings demonstrate that ML-1 is a novel cytokine with a distinct function, and suggest a different receptor for ML-1 on PBE cells.

Topics: Amino Acid Sequence; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Humans; Intercellular Adhesion Molecule-1; Interleukin-17; Interleukin-6; Interleukin-8; Molecular Sequence Data; Plant Proteins; Pollen; Respiratory Mucosa; Sequence Homology, Amino Acid; Tissue Distribution

Asthma is associated with increased production of IL-4 and IL-13.. Because many of the effects of these cytokines are mediated by activation of signal transducer and activator of transcription 6 (STAT-6), we investigated expression and function of this transcription factor in the airways.. STAT-6 expression was investigated through use of immunohistochemistry or RT-PCR applied to bronchial biopsy specimens or brushings from normal control or asthmatic subjects. STAT-6 function was investigated by means of Western blotting and ELISA applied to primary epithelial cell cultures.. Immunohistochemistry revealed that the bronchial epithelium was the major site of STAT-6 expression, both cytoplasmic and nuclear staining being observed. The level of STAT-6 expression in subjects with mild asthma (median [range] percent epithelial staining, 3.4% [0% to 16.0%]; n = 14) did not differ significantly from that in normal controls (4.7% [0.0% to 20.0%]; n = 11); however, in subjects with severe asthma, epithelial STAT-6 expression (13.7% [4.8% to 25.7%]; n = 9) was increased in comparison with subjects with mild asthma and normal controls (P < .05). RT-PCR analysis showed that epithelial STAT-6 expression was heterogeneous and comprised both full-length STAT-6 and the dominant-negative variant that lacks the SH2 domain. Treatment of primary cultures of bronchial epithelial cells with IL-4 resulted in STAT-6 phosphorylation and stimulation of IL-8 secretion; however, no difference in the responses of epithelial cells was observed between normal (n = 12) and asthmatic (n = 14) donors.. These data demonstrate expression and activation of STAT-6 in normal and asthmatic bronchial epithelium. The activity of this transcription factor is likely to play a key role in mediating the responses of the bronchial epithelium to T(H)2 cytokines that are characteristic of the asthmatic phenotype.

Topics: Asthma; Bronchi; Cells, Cultured; Humans; Immunohistochemistry; Interleukin-13; Interleukin-4; Interleukin-8; Janus Kinase 1; Models, Biological; Phosphorylation; Protein-Tyrosine Kinases; Respiratory Mucosa; RNA, Messenger; STAT6 Transcription Factor; Trans-Activators; Transcription, Genetic

Mast cells produce substances with antiinflammatory properties in addition to their capacity to release proinflammatory mediators. To further probe the antiinflammatory aspect of mast-cell function we investigated the ability of human mast cells (huMCs) to produce interleukin (IL)-1 receptor antagonist (IL-1ra) in response to high-affinity Fc receptor for immunoglobulin E (Fcalpha RI) aggregation, and examined IL-1ra in bronchoalveolar lavage fluid (BALF) to determine whether it might be of mast-cell origin. Using a ribonuclease protection assay, flow cytometry, and enzyme-linked immunosorbent assay (ELISA), IL-1ra message and protein were found to be constitutively expressed in cultured huMCs. Upon stimulation through Fcalpha RI, IL-1ra message was upregulated in huMCs and IL-1ra protein secreted from cultured huMCs and isolated human lung mast cells. By immunoblot analysis, huMCs were found to produce the 17-kD form of IL-1ra and the presence of IL-1ra in human lung mast cells was confirmed by immunohistochemistry. In BALF obtained from allergic asthmatic subjects, IL-1ra production increased after specific antigen challenge, with the 17-kD isoform of IL-1ra predominating. These findings demonstrate that huMCs produce and release IL-1ra after Fcalpha RI aggregation, which may contribute to a local inhibition of IL-1-dependent effects on inflammation in the lung.

Topics: Allergens; Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Bronchoscopy; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Flow Cytometry; Gene Expression Regulation; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Immunologic Capping; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Interleukin-8; Lung; Mast Cells; Receptors, IgE; Receptors, Interleukin-1; RNA, Messenger; Secretory Rate; Sialoglycoproteins

IL-15 is a T(H)1-related cytokine that shares many biologic activities with IL-2. Both cytokines bind a specific alpha subunit, and they share the same beta and gamma common receptor subunits for signal transduction. IL-15 has recently been shown to be upregulated in T cell-mediated inflammatory disorders, such as rheumatoid arthritis and inflammatory bowel diseases. However, the role and expression of IL-15 in inflammatory lung disease has not been investigated.. In the present study we have evaluated the expression of IL-15 mRNA and protein in bronchial biopsy specimens obtained from patients with sarcoidosis (n = 8), tuberculosis (n = 7), chronic bronchitis (n = 10), and bronchial asthma (n = 8) and compared its expression with that seen in normal control subjects (n = 11).. In situ hybridization and immunocytochemistry were used to detect the number of cells expressing IL-15 mRNA and protein, respectively, within sections of bronchial tissues from all subject groups. In addition, double immunocytochemistry was used to characterize the cellular source of IL-15.. The number of IL-15(+) cells was significantly higher within tissue from patients with sarcoidosis, tuberculosis, and chronic bronchitis compared with that in asthmatic patients and normal control subjects. Similar results were obtained for IL-15 immunoreactivity by using immunohistochemistry. Furthermore, double immunostaining revealed that neutrophils and macrophages are the major source of IL-15.. These results suggest that the expression of IL-15 may be associated with T(H)1-mediated chronic inflammatory diseases of the lung.

Topics: Asthma; Bronchitis; Humans; Immunohistochemistry; Interleukin-15; Interleukin-8; Neutrophils; RNA, Messenger; Sarcoidosis; Th1 Cells; Tuberculosis, Pulmonary

Cigarette smoking is common in asthmatic patients, and we investigated the impact of cigarette smoking on airway inflammation in asthma.. Single-center observational study of airway inflammation in asthmatic and healthy smokers and nonsmokers.. Asthma research unit in a university hospital.. Sixty-seven asthmatic and 30 nonasthmatic subjects classified as smokers or nonsmokers. Asthmatics had chronic, stable asthma and were not receiving inhaled or oral steroids at the time of the study.. We examined induced-sputum cell counts and levels of interleukin (IL)-8 and eosinophilic cationic protein (ECP). Bronchial hyperreactivity was assessed using methacholine challenge.. Asthmatic smokers had higher total sputum cell counts than nonsmoking asthmatics and both smoking and nonsmoking healthy subjects. Smoking was associated with sputum neutrophilia in both asthmatics and nonasthmatics (median, 47% and 41%, respectively) compared with nonsmokers (median, 23% and 22%, respectively), and sputum IL-8 was increased in smokers compared with nonsmokers, both in subjects with asthma (median, 945 pg/mL vs 660 pg/mL, respectively) and in healthy subjects (median, 1,310 pg/mL vs 561 pg/mL, respectively). Sputum eosinophils and ECP levels were higher in both nonsmoking and smoking asthmatics than in healthy nonsmokers. In smoking asthmatics, lung function (FEV(1) percent predicted) was negatively related to both sputum IL-8 (r = - 0.52) and sputum neutrophil proportion (r = - 0.38), and sputum IL-8 correlated positively with smoking pack-years (r = 0.57) and percent neutrophil count (r = 0.51).. In addition to the eosinophilic airway inflammation observed in patients with asthma, smoking induces neutrophilic airway inflammation; a relationship is apparent between smoking history, airway inflammation, and lung function in smoking asthmatics.

Topics: Adult; Asthma; Blood Proteins; Eosinophil Granule Proteins; Eosinophils; Female; Forced Expiratory Volume; Humans; Interleukin-8; Leukocyte Count; Male; Middle Aged; Neutrophils; Reference Values; Ribonucleases; Smoking; Sputum

To investigate the role of neutrophils and interleukin-8 (IL-8) in pathogenesis of asthma.. Pulmonary function and bronchial provocation test were performed to identify and evaluate asthma. Superoxide anion O(2)(-.) generation of peripheral blood neutrophils, IL-8 and malondialhyde (MDA) levels in plasma and induced sputum were measured.. The O(2)(-.) generation [(20.9 +/- 5.1) nmol] of peripheral blood neutrophils from moderate-severe asthmatics were significantly higher than that [(15.2 +/- 4.2) nmol] from mild asthmatics (P < 0.01), in which that was still significantly higher than that [(11.3 +/- 2.4) nmol] from normal subjects (P < 0.05). The O(2)(-.) generation of peripheral blood neutrophils had significantly negative correlation with PD(20)FEV(1) (r = -0.693, P < 0.05). The levels of IL-8 [(585 +/- 75) ng/L, (791 +/- 103) ng/L] and MDA levels [(6.3 +/- 1.6) mmol/L, (21.8 +/- 6.3) mmol/L] in plasma and induced sputum from unstable asthmatics were higher than those [(227 +/- 54) ng/L, (322 +/- 95) ng/L and (5.4 +/- 1.0) mmol/L, (15.1 +/- 5.6) mmol/L] from stable asthmatics (P < 0.01), more over, in the latter group, they were still significantly higher than those (188 +/- 46) ng/L, (224 +/- 51) ng/L and [(4.1 +/- 0.4) mmol/L, (9.5 +/- 4.2) mmol/L] from normal subjects (P < 0.05). The levels of MDA in the induced sputum from stable asthmatics showed a negative correlation with PD(20)FEV(1) (r = -0.708, P < 0.01). The levels of IL-8 in the induced sputum showed a positive correlation with percentage of neutrophils in the induced sputum (r = 0.838, P < 0.01).. IL-8 and oxygen free radicals generated by neutrophils might participate in development of airway inflammation and airway hyperresponsiveness in asthma.

Topics: Adult; Asthma; Female; Humans; Interleukin-8; Lung; Male; Malondialdehyde; Neutrophils; Sputum; Superoxides

There is abundant evidence for T-lymphocyte recruitment into the airways in allergic inflammatory responses. This study has tested the hypothesis that T-cell chemotaxis induced by platelet-activating factor (PAF) and human recombinant interleukin-8 (hrIL-8) can be attenuated by inhibition of phosphodiesterase activity and raised intracellular 3',5'-cyclic adenosine monophosphate (cAMP) levels. This study used theophylline, a nonselective phosphodiesterase (PDE) inhibitor, and rolipram, a selective PDE4 inhibitor, to study the effect of PDE inhibition on T-cell chemotaxis. The beta2-adrenoceptor agonist, salbutamol, the adenylyl cyclase activator, forskolin, and the cAMP analogue, dibutyryl cAMP (db-cAMP), were used to demonstrate a role for raised cAMP levels. T-cells were obtained from 10 atopic asthmatics, and the phenotype of migrating cells was examined by flow cytometry. Theophylline caused an inhibition of both PAF-and hrIL-8-induced chemotaxis (mean+/-SEM maximum inhibition at 1 mM: 73+/-4% and 48+/-8% for hrIL-8 and PAF, respectively) that was not specific for the CD4+, CD8+, CD45RO+ or CD45RA+ T-cell subsets. T-cell chemotaxis was more sensitive to treatment with rolipram whose effect was already significant from 0.1 microM on hrIL-8-induced chemotaxis. Both a low concentration of salbutamol (0.1 mM) and forskolin (10 microM) potentiated the inhibitory effect of a low concentration of theophylline (25 microM) on responses to PAF but not to hrIL-8. Finally, T-cell chemotaxis was also inhibited by db-cAMP. It is concluded that attenuation of T-cell chemotaxis to two chemoattractants of relevance to asthma pathogenesis can be achieved via phosphodiesterase inhibition and increased intracellular 3', 5'-cyclic monophosphate using drugs active on cyclic nucleotide phosphodiesterase. This action may explain the anti-inflammatory effects of theophylline and related drugs in asthma.

Topics: Albuterol; Asthma; Bucladesine; Chemotaxis, Leukocyte; Colforsin; Cyclic AMP; Humans; Interleukin-8; Phosphodiesterase Inhibitors; Phosphoric Diester Hydrolases; Platelet Activating Factor; Rolipram; T-Lymphocytes; Theophylline

Although airway inflammation is recognized as a key feature of asthma, the characteristics of airway inflammation in children with acute severe asthma are not well defined. The aim of this study was to describe the characteristics of airway inflammation in children with an acute exacerbation of asthma using sputum cell counts and fluid-phase measurements and to examine the changes in these parameters upon resolution of the exacerbation. Children (n = 38) presenting to the Emergency Department with acute asthma underwent successful sputum induction using ultrasonically nebulized normal saline (n = 22), or expectorated sputum spontaneously (n = 16). Sputum induction was repeated at least 2 wk later when the children had recovered (n = 28). Sputum portions were selected, dispersed and total and differential cell counts performed. Neutrophil elastase and EG2-positive eosinophils were assessed and fluid-phase eosinophil cationic protein (ECP), myeloperoxidase (MPO), interleukin-8 (IL-8), and IL-5 were measured. During the acute exacerbation the median (range) total cell count was 8.4 x 10(6)/ml (0.5 to 190.3), and fell significantly at resolution to 1.3 x 10(6)/ml (p < 0.01). The inflammatory cell infiltrate was mixed and included eosinophils (0.8 x 10(6)/ml), neutrophils (3.3 x 10(6)/ml), and mast cells. EG2(+) cells were high and correlated with the degree of airflow obstruction (r = -0.5, p = 0.02). They decreased significantly at resolution as did supernatant ECP (1,078 versus 272 ng/ml), suggesting that eosinophils were activated during the exacerbation. MPO was 220 ng/ ml at exacerbation and fell significantly to 1 ng/ml at resolution. Levels of IL-8 and IL-5 were elevated during the acute exacerbation and IL-8 concentrations decreased at resolution. In conclusion, airway inflammation can be studied in children with acute asthma by sputum induction. Airway inflammation is present during an acute exacerbation of asthma, and is characterized by infiltration and activation of both eosinophils and neutrophils. The heterogeneity of airway inflammation in acute asthma may influence response to corticosteroid therapy.

Topics: Acute Disease; Adolescent; Asthma; Child; Eosinophils; Female; Humans; Interleukin-8; Male; Neutrophil Infiltration; Respiratory Tract Infections; Sputum

Cough persisting after a respiratory infection is common in children and is often managed as asthma. However, little is known about the pathophysiologic mechanisms of such cough and how it compares with asthma.. We used the technique of induced sputum to examine the inflammatory index values associated with persistent cough or allergic asthma in children. We hypothesized that the sputum from children with persistent postinfectious cough would differ from that of children with allergic asthma in that the former would lack eosinophils compared with the latter.. Sputum production was induced with hypertonic saline solution in 34 children: 12 with cough persisting for 1 month or more after an apparent respiratory tract infection, not treated with corticosteroid; 11 with untreated atopic asthma, not using inhaled corticosteroid; and 11 with treated atopic asthma using inhaled corticosteroid.. The percentage of eosinophils in the sputum of children with cough was significantly lower than in the sputum of children with untreated allergic asthma (median 0.5% vs 14.5%, P <.0001). Similarly, the percentage of eosinophils in the sputum of children with asthma treated with inhaled steroids was significantly lower compared with untreated asthmatic children (1.5% vs 14.5%, P <.0001). The peripheral blood eosinophils, serum eosinophil cationic protein, and nasal percent eosinophils of the patients with cough were also significantly lower than those from patients with untreated asthma. Methacholine challenge in 6 of the 11 cough patients tested showed mild-to-moderate hyperresponsiveness, whereas the other 5 had a negative methacholine challenge.. Children with persistent postinfectious cough do not have airway eosinophilia typical of untreated asthma. Despite the absence of eosinophilic inflammation, some of the patients with chronic cough had reactive airways. These results suggest that postinfectious cough in children has different pathophysiologic features than allergic asthma and probably represents a different disease.

Topics: Asthma; Blood Proteins; Child; Child, Preschool; Cough; Eosinophil Granule Proteins; Eosinophils; Female; Humans; Infections; Inflammation Mediators; Interleukin-8; Leukocyte Count; Male; Methacholine Compounds; Ribonucleases; Sputum

The inflammatory events in the airways at the time of acute respiratory failure from acute severe asthma are poorly understood. To determine the patterns of cellular inflammation in the airways in acute severe asthma, we analyzed tracheal aspirates collected within 12 h of intubation from patients intubated emergently for acute severe asthma (n = 10) and from patients intubated electively for nonpulmonary surgery (n = 14). The number of neutrophils in tracheal aspirates from asthma patients was 10 times higher than normal (4.2 [0.6 to 335.0] [median, range] versus 0.4 [0.009 to 9.4] x 10(6)/ml, p = 0.001), and there was a strong trend for a positive relationship between neutrophil number and duration of intubation (r(s) = 0.64, p = 0.06). Although eosinophil numbers were also significantly higher than normal (0.5 [0.0 to 23.3] versus 0.0 [0.0 to 0.1] x 10(6)/ml, p = 0.003), the numbers of eosinophils were 8-fold less than neutrophils, and there was no significant correlation between eosinophil number and duration of intubation (r(s) = 0.4, p = 0.26). Interleukin-8 (IL-8), a chemoattraction for neutrophils, was 19 times higher than normal in tracheal aspirates from asthmatic patients (75.0 [9.0 to 168.0] versus 4.0 [0.08 to 24.0] ng/ml, p < 0. 05) and correlated significantly with the neutrophil number (r(s) = 0.77, p = 0.03). Furthermore, the IL-8 levels correlated positively with the duration of mechanical ventilation (r(s) = 0.74, p = 0.03). Surprisingly, the number of neutrophils increased significantly during the period of intubation in the asthmatic subjects, possibly because of intravenous corticosteroid treatment. We conclude that neutrophils are the dominant inflammatory leukocyte characterizing airway inflammation in acute severe asthma that requires mechanical ventilation, and that IL-8 is an important mediator of this neutrophilia.

Topics: Adult; Aged; Asthma; Female; Humans; Inflammation Mediators; Interleukin-8; Intubation, Intratracheal; Leukocyte Count; Male; Middle Aged; Mucus; Neutrophils; Respiration, Artificial; Trachea

We have previously reported that human airway smooth-muscle (ASM) cells produce abundant interleukin (IL)-8, a major neutrophil chemoattractant involved in asthma exacerbations. Here, we tested the effects of the beta(2)-agonists salbutamol (Salbu) and salmeterol (Salme) on IL-8 release and tumor necrosis factor (TNF)-alpha-induced IL-8 release from ASM cells. We found that TNF-alpha strongly enhanced IL-8 release in a time- and concentration-dependent manner, whereas Salbu, Salme, the direct adenylyl cyclase activator forskolin (FSK), and the cyclic monophosphate (cAMP) analogue 8-bromoadenosine 3',5'-cAMP (8-Br-cAMP) alone weakly stimulated IL-8 release. TNF-alpha (10 ng/ml)-induced IL-8 release was markedly inhibited by the steroids dexamethasone (Dex) (0.1 to 10 microM) and fluticasone (Flut) (0.01 to 1 microM) but unaffected by Salbu, Salme, FSK, or 8-Br-cAMP. However, a combination of Dex (1 microM) or Flut (0.1 microM) with Salbu (10 microM), Salme (1 microM), FSK (10 microM), or 8-Br-cAMP (10 and 100 microM) significantly enhanced the inhibition by Dex or Flut alone. Experiments with KT5720, a selective inhibitor of cAMP-dependent protein kinase A; rolipram, a selective inhibitor of type IV phosphodiesterase; and ICI-118,551, a beta(2)-receptor antagonist, suggested that the synergistic inhibition was mediated by beta(2)-receptor in a cAMP-dependent manner. This novel synergistic interaction of beta(2)-agonists and steroids may partly explain the benefits that result when these agents are combined to treat asthma.

Topics: 8-Bromo Cyclic Adenosine Monophosphate; Adrenal Cortex Hormones; Adrenergic beta-2 Receptor Antagonists; Adrenergic beta-Agonists; Adrenergic beta-Antagonists; Albuterol; Androstadienes; Asthma; Cell Survival; Cells, Cultured; Colforsin; Cyclic AMP-Dependent Protein Kinases; Dexamethasone; Drug Synergism; Fluticasone; Humans; Interleukin-8; Muscle, Smooth; Phosphodiesterase Inhibitors; Propanolamines; Receptors, Adrenergic, beta-2; Respiratory System; Rolipram; Salmeterol Xinafoate; Trachea; Tumor Necrosis Factor-alpha

Patients with glucocorticoid (GC)-dependent asthma present an ongoing inflammation of the airways despite chronic long-term treatment with oral GC. Interleukin (IL)-8 and granulocyte/macrophage colony-stimulating factor (GM-CSF) have been implicated in airway inflammation in severe asthma and their synthesis is normally repressed by GC. To further characterize the inflammatory process in GC-dependent asthma, we measured the release of IL-8 and GM-CSF by peripheral blood mononuclear cells (PBMC) of eight normal subjects, six untreated controlled asthmatics, six untreated uncontrolled asthmatics, and nine GC-dependent asthmatics. We show that PBMC from GC-dependent asthmatics released high amounts of these cytokines despite chronic in vivo exposure to GC (p < 0.001 versus normal subjects). In contrast, when untreated uncontrolled asthmatics were given a short course of oral GC, IL-8 and GM-CSF production was inhibited (p = 0.0078). Release of IL-8 and GM-CSF by PBMC of GC-dependent asthmatics was reduced after in vitro GC treatment (p < 0.002). We investigated whether the incapacity of GC to inhibit production of these cytokines in vivo was the result of a dysregulation of the glucocorticoid receptor (GR) in GC-dependent asthma. GRalpha and GRbeta are, respectively, the functional receptor and a putative dominant negative form of the receptor. Western blot and polymerase chain reaction (PCR) analyses indicated that GRalpha was expressed at similar level in all groups and was largely predominant over GRbeta. Thus, persistent release of IL-8 and GM-CSF in GC-dependent asthma is not associated with low expression of GRalpha or overexpression of GRbeta.

Topics: Adult; Asthma; Female; Glucocorticoids; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Receptors, Glucocorticoid; RNA, Messenger; Severity of Illness Index

The alveolar macrophage (AM), a major defense cell in the lung, participates in immune and inflammatory reactions through the release of several regulatory and chemotactic cytokines. In particular, macrophages are considered to play a pivotal proinflammatory role in the production and maintenance of airway inflammation and bronchial hyperreactivity. To assess the phenotypic pattern of AM from asthmatic subjects, we performed the following experiments: 1) cytofluorometric analysis of specific phenotypic features (CD11b, CD14, CD16, CD45, HLA-DR, CD71, CD95, and CD44) 2) assessment of the production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1, and the chemotactic regulatory cytokine IL-8 by unstimulated and lipopolysaccharide-stimulated AM. In these patients, we phenotypically characterized the AM, showing their strong proinflammatory activity also in patients with mild asthma. Their activity has been clarified by our biomolecular data that showed a constitutive basal IL-8 production by AM, and also indicated that IL-1 and TNF-alpha were able to upregulate the ability of activated human AM to produce IL-8 at the protein and messenger ribonucleic acid (mRNA) levels.

Topics: Asthma; Cytokines; Gene Expression Regulation; Humans; Interleukin-1; Interleukin-8; Lipopolysaccharides; Macrophage Activation; Macrophages, Alveolar; Monocytes; Phenotype; RNA, Messenger; Transcription, Genetic; Transcriptional Activation; Tumor Necrosis Factor-alpha

A series of bis(trifluoromethyl)pyrazoles (BTPs) has been found to be a novel inhibitor of cytokine production. Identified initially as inhibitors of IL-2 synthesis, the BTPs have been optimized in this regard and even inhibit IL-2 production with a 10-fold enhancement over cyclosporine in an ex vivo assay. Additionally, the BTPs show inhibition of IL-4, IL-5, IL-8, and eotaxin production. Unlike the IL-2 inhibitors, cyclosporine and FK506, the BTPs do not directly inhibit the dephosphorylation of NFAT by calcineurin.

Topics: Animals; Asthma; Cell Division; Chemokine CCL11; Chemokines, CC; Combinatorial Chemistry Techniques; Cyclosporine; Cytokines; DNA-Binding Proteins; Genes, Reporter; Haplorhini; Humans; Immunosuppressive Agents; In Vitro Techniques; Interleukin-2; Interleukin-4; Interleukin-5; Interleukin-8; Jurkat Cells; Leukocytes, Mononuclear; Luciferases; NFATC Transcription Factors; Nuclear Proteins; Protein Synthesis Inhibitors; Pyrazoles; Rats; Transcription Factors

Floating fog occurs every summer in Kushiro City in Japan, and the annual average of fog water pH in the past 4 years has been under 5.0. We previously reported that epidemiologically fog was the most important positive factor contributing to increased hospital visits of asthmatic patients compared with other meteorological values and air pollutants. This study aimed to investigate the mechanism of the effects of naturally-occurring acid fog on asthmatic patients. We compared pulmonary functions and inflammatory mediators in induced sputum between the foggy (July 1995) and the non-foggy (May 1996) season, and assessed airway responsiveness to hypo-osmolar aerosol. Forty-four out of 118 asthmatic patients of Kushiro City residents participated, pulmonary function tests were completed in 36 patients, and sputum data were available in 26 patients in both seasons. Percent forced expiratory volume in 1 sec (FEV1) was significantly (P< 0.05) decreased, and % peak expiratory flow rate (PEFR) had a trend to decrease in the foggy season more than in the non-foggy, and sputum eosinophil cationic protein (ECP) and interleukin (IL)-8 were higher in the foggy season but not significantly. A moderate inverse correlation was revealed between sputum ECP and %PEFR in the foggy season (r= -0.55, P<0.005). Subjects were divided into two groups according to the best PEFR; one had >10% lower PEFR levels in the foggy season than in the non-foggy season (Group A, n = 7), the remainder did not (Group B, n = 19). In group A, sputum ECP was significantly increased (P< 0.01) in the foggy season, but there were no changes in IL-8 and prostaglandin D2. Ultrasonic nebulized distilled water provocation test revealed no differences between group A and B. These results suggested that eosinophilic inflammation rather than hypo-osmolar effect of fog might contribute to respiratory deterioration by inhalation of naturally-occurring acid fog.

Topics: Acid Rain; Adult; Aged; Asthma; Blood Proteins; Eosinophil Granule Proteins; Female; Forced Expiratory Volume; Humans; Interleukin-8; Japan; Male; Middle Aged; Peak Expiratory Flow Rate; Prostaglandin D2; Ribonucleases; Sputum; Weather

Cytokines are considered to play a role in the airway inflammation of bronchial asthma. We examined the cellular profile and cytokine levels in induced sputum samples obtained before and after treatment with beclomethasone dipropionate (BDP, 800 microg/day, for 4 weeks) in 12 mild to moderate asthmatic subjects who had not previously received inhaled glucocorticosteroids. Sputum was induced with a 20-min inhalation of 3% saline by an ultrasonic nebulizer. The freshly expectorated sputum separated from the saliva was analyzed for cell counts, for the concentration of interleukin-8 (IL-8), and for the concentration of granulocyte-macrophage colony-stimulating factor (GM-CSF). The mean percentage of eosinophils in the sputum samples decreased significantly after BDP treatment, but no significant change in the percentage of neutrophils was observed. The mean IL-8 and GM-CSF levels also decreased significantly after treatment. The BDP treatment was associated with an increase in the mean peak expiratory flow (PEF) and with a decrease in the diurnal variation of PEF. These results suggest that inhaled steroids improve airway inflammation and lung function in asthmatics, presumably in part by inhibiting the synthesis of inflammatory cytokines such as IL-8 and GM-CSF.

Topics: Administration, Inhalation; Administration, Topical; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Beclomethasone; Cell Count; Eosinophils; Female; Glucocorticoids; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Male; Middle Aged; Neutrophils; Peak Expiratory Flow Rate; Reproducibility of Results; Spirometry; Sputum

To study whether the individual inflammatory response to ozone was reproducible, dose-dependent, and time-dependent, we performed two exposures to 250 ppb ozone, one to 125 ppb and one to filtered air, each for 3 h of intermittent exercise and separated by at least 1 wk. Twenty-one healthy and 15 asthmatic subjects participated in the study. One hour after the two exposures to 250 ppb ozone we observed a mean increase in sputum neutrophils of 17.9 and 17.9% in healthy and of 20.3 and 15.2% in asthmatic subjects (p < 0.05 each). Twenty-four hours after exposure, the respective values were 11.9 and 14.8%, and 9.1 and 16.1% (p < 0.05 each). In the whole group of subjects, individual changes in the percentage of neutrophils were significantly correlated between the two exposure days 1 h (r = 0.87, p < 0.001; intraclass correlation coefficient [Ri] = 0.86) as well as 24 h (r = 0.79, p < 0.001; Ri = 0.71) after exposure. The percentages of lymphocytes were increased 24 h after exposures (all subjects combined: p < 0.05). The decrease in FEV1 in both groups (p < 0.01), was also reproducible (r = 0.77, p < 0.001), but there were no correlations between changes in sputum parameters and lung function. Exposure to 125 ppb ozone caused a small increase (p < 0. 05) in the percentage of neutrophils in asthmatic subjects and in the concentrations of interleukin-8 in both groups combined. Our data demonstrate that inflammatory and lung function responses to ozone differ between individuals and are reproducible but not related to each other. Therefore, these responses appear to represent two independent factors underlying the airway response to ozone.

Topics: Adult; Air Pollutants; Asthma; Cell Count; Dose-Response Relationship, Drug; Eosinophils; Female; Forced Expiratory Volume; Humans; Interleukin-8; Lymphocytes; Macrophages; Male; Middle Aged; Neutrophils; Ozone; Physical Exertion; Reproducibility of Results; Respiratory System; Sputum; Vital Capacity

Soot particles, asbestos fibres and irritant gas are common air pollutants which are able to induce lung and airway pulmonary injury. The aim of this study was to investigate the effect of a simultaneous NO2 and particle or fibre exposure on the proinflammatory specific mRNA expression and protein secretion of human alveolar macrophages (AM) in comparison to only particle or fibre exposed AM. AM were simultaneously exposed to FR 101, P 90, TiO2 or Chrysotile B at a concentration of 100 microg/10(6) cells and to NO2 at a concentration of 1.0 ppm for 30 min. Particle or fibre exposure of the AM was continued in humidified air at 5% CO2 and 37 degrees C for an additional hour (harvesting of total RNA) or additional 7 hrs (harvesting of culture supernatant). The mRNA expression of the proinflammatory cytokines IL-1beta, IL-6, IL-8 and TNF-alpha of NO2-particle/fibre co-exposed AM and only particle or fibre exposed AM was detected using specific RT-PCR. IL-1beta-, IL-6-, IL-8- and TNF-alpha-specific protein secretion was measured by ELISA. Cytotoxicity was detected by lactatedehydrogenase quantification in the culture supernatant. We observed an increased IL-1beta-, IL-6-, IL-8- and TNF-alpha-specific mRNA expression of particle or fibre exposed AM, which was decreased after an additional NO2 exposure. Also the particle or fibre exposure induced significant increase in IL-1beta-, IL-6-, IL-8 and TNF-alpha-release of AM which was decreased after an additional NO2 exposure (p <0.031). The relative cytotoxicity of the NO2-particle/fibre co-exposure was higher than the particle or fibre induced cytotoxicity, but mostly <10%. Therefore it is concluded that particle or fibre exposure may result in an increase in proinflammatory cytokine release by AM, which may be decreased by toxic NO2 due to the oxidative potential (e.g. lipidperoxydation) of this irritant gas. Particle, asbestos fibre and irritant gas exposure may induce airway and pulmonary injury by the activation of AM and consecutive proinflammatory cytokine release.

Topics: Aged; Air Pollutants; Asbestos, Serpentine; Asthma; Bronchial Neoplasms; Bronchoalveolar Lavage Fluid; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Cells, Cultured; Cytokines; Drug Synergism; Female; Gene Expression Regulation; Humans; Inflammation; Interleukin-1; Interleukin-6; Interleukin-8; Irritants; Lung Neoplasms; Macrophages, Alveolar; Male; Middle Aged; Nitrogen Dioxide; Particle Size; RNA, Messenger; Titanium; Tumor Necrosis Factor-alpha

Epithelial hyperplasia and mucosal infiltration of leukocytes are common features of chronic rhinosinusitis. The epithelium can produce chemoattractant cytokines that may contribute to leukocyte infiltration in rhinosinusitis.. We sought to determine whether mucosal IL-8 gene expression is increased in chronic rhinosinusitis and to relate IL-8 gene expression to disease severity.. We used both a noncompetitive and a quantitative, competitive reverse transcription-polymerase chain reaction to examine IL-8 gene expression in samples of sinus mucosal tissue obtained during surgery from 22 patients with chronic rhinosinusitis and 9 normal control subjects. IL-8 gene expression was related to disease severity assessed by sinus computed tomography (CT) scores and to symptom scores assessed by means of a questionnaire.. Sinus mucosal IL-8 gene expression was not detected in any of the control subjects but was present in 12 of 22 (55%) patients with rhinosinusitis. Sinus CT scores and symptom scores were both significantly higher in patients with positive mucosal IL-8 gene expression than in subjects with no detectable IL-8 gene expression. Positive IL-8 gene expression was not predicted by history of prior surgery nor by atopic or asthmatic status. In 9 subjects with positive IL-8 gene expression, levels of mRNA expression, assessed by competitive reverse transcription-polymerase chain reaction, correlated significantly (rho = 0.72, P <.05) with sinus CT scores.. Sinus mucosal expression of the gene for IL-8 is increased in patients with chronic rhinosinusitis, and the level of IL-8 gene expression correlates with disease severity.

Topics: Adult; Aged; Asthma; Chronic Disease; Epithelial Cells; Female; Gene Expression; Humans; Hypersensitivity, Immediate; Interleukin-8; Male; Middle Aged; Mucous Membrane; Reverse Transcriptase Polymerase Chain Reaction; Rhinitis; RNA, Messenger; Sinusitis; Tomography, X-Ray Computed

Bronchoalveolar lavage (BAL) fluid from patients with birch-pollen allergy lavaged during the season showed an elevated chemotactic activity for eosinophils compared with BAL fluid from the same patients before the start of the season.. The aim of this study was to identify the eosinophil chemotactic agents in the BAL fluid, to compare these findings with in vitro studies on selected cytokines, and to investigate the interactions between these cytokines.. Neutralizing antibodies for interleukins (IL) -2, -5 and -8, RANTES and leukaemia inhibitory factor (LIF) were added to the BAL fluid, and the chemotactic activity was tested with eosinophils from allergic donors. Eosinophils from healthy donors were preincubated with IL-5 in order to mimic the primed state of eosinophils from allergics, and the migration towards recombinant IL-5, IL-8, and RANTES in different combinations was measured. Eosinophils from allergic donors were also used.. Anti-IL-5, anti-IL-8 and anti-RANTES inhibited the chemotactic activity in the BAL fluid. Recombinant RANTES induced migration, which was enhanced by preincubation of the cells with IL-5. Only eosinophils from symptomatic allergics responded to IL-8, and IL-5 was not sufficient to prime normal eosinophils in vitro to an IL-8 response. A negative correlation was found between the level of in vivo activation of the cells and their response to IL-5, and a positive correlation with the response to RANTES.. IL-8 and RANTES are important for eosinophil accumulation to the lung of pollen-allergic asthmatics. IL-5 alone may not be responsible for the priming of eosinophils in vivo, but is an essential cofactor for the other chemoattractants.

Topics: Adult; Asthma; Chemokine CCL5; Chemotaxis, Leukocyte; Eosinophils; Female; Humans; Interleukin-5; Interleukin-8; Lung; Male

Segmental allergen challenge is a powerful tool to study inflammatory reactions in asthmatic airways. There is little information on the early events at 5 min and 4 h after allergen challenge with respect to the cell influx and the chemokine interleukin-8 (IL-8).. Seven mild to moderate allergic asthmatics (AA group), 5 allergic nonasthmatics (ANA group) and 5 nonallergic controls underwent segmental allergen challenge, with allergen doses based upon skin reactivity. Bronchoalveolar lavage (BAL) samples were obtained before, 5 min and 4 h postchallenge, and were analyzed for cell numbers and differential counts, eosinophil and neutrophil chemotactic activity, and levels of IL-8.. At 5 min postchallenge, no changes were observed compared to baseline. At 4 h postchallenge, an increase was found in the number of neutrophils and the levels of IL-8, which was dependent on the dose of allergen in the AA and ANA group. At the same allergen dose, the increases in neutrophils and levels of IL-8 were calculated to be 91 and 67 times higher, respectively, in AA than in ANA. Levels of IL-8 correlated with the number of neutrophils and with the in vitro neutrophil chemotactic activities in BAL fluid.. Neutrophil chemotactic activity is increased in BAL fluid at 4 h after segmental allergen challenge. We suggest that apart from IgE-mediated mast cell degranulation, additional local factors in the airways determine the degree of IL-8 increase and neutrophil influx.

Topics: Adult; Allergens; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Cell Count; Chemotaxis, Leukocyte; Eosinophils; Female; Humans; Hypersensitivity; Interleukin-8; Lipopolysaccharides; Male; Neutrophils; Skin Tests

Recent observations show that atopic asthmatic subjects have increased sensitivity to respirable endotoxin (or LPS) compared with normal persons. In vitro studies demonstrate that LPS enhances eosinophil survival. These observations suggest that the effects of inhaled LPS in asthmatic subjects may include increases in the number of airway eosinophils.. We sought to determine whether low-level nasal LPS challenge causes an increase in eosinophil numbers in the nasal airways of atopic or normal subjects.. Sixteen volunteers (10 atopic asthmatic subjects and 6 normal subjects) underwent 2 nasal challenge sessions. In one session, one nostril was challenged with saline and the other with 0. 1 microg of LPS. During the second session, 0.3 microg and 1.0 microg of LPS was delivered to each nostril, respectively. Nasal lavage fluid was obtained from each nostril before challenge, as well as 4 and 24 hours after challenge, and examined for the percent of total cells that were eosinophils and neutrophils, as well as cytokine levels.. LPS (1.0 microg) increased the percent of eosinophils in nasal lavage fluid 4 hours after challenge in atopic subjects only. There was also a correlation between constitutive nasal GM-CSF and eosinophil response to LPS in atopic subjects.. LPS challenge increases eosinophils in the airways of atopic subjects.

Topics: Adolescent; Adult; Asthma; Blood Proteins; Cell Movement; Eosinophil Granule Proteins; Eosinophils; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Granulocytes; Humans; Hypersensitivity, Immediate; Inflammation Mediators; Interleukin-8; Lipopolysaccharides; Male; Middle Aged; Nasal Mucosa; Nasal Provocation Tests; Pilot Projects; Ribonucleases

The immunopathological mechanism for occupational asthma induced by toluene diisocyanate (TDI) remains to be further clarified. There have been few reports suggesting involvement of neutrophils in inducing bronchoconstriction after TDI inhalation.. To further understand the role of neutrophils in the pathogenesis of TDI-induced asthma.. Eight TDI-induced asthmatic subjects were classified as group I, and five exposed workers who had complained of work-related symptoms and worked in the same workplace, but showed negative bronchial challenges were enrolled as controls (group II). Serum neutrophil chemotactic activity during TDI bronchial challenge test was measured by the Boyden chamber method. Induced sputum was collected before and after the TDI bronchial challenge test. The myeloperoxidase (MPO) and interleukin (IL) -8 levels in the sputum were measured using RIA and ELISA.. Serum neutrophil chemotactic activity significantly increased at 10 min (P = 0.01), then decreased at 60 min (P = 0.02) and remained unchanged for up to 420 min (P = 0.07) in group I subjects, while no significant changes were found in group II subjects (P > 0.05). MPO and IL-8 were abundantly present in the sputum of all the TDI-induced asthmatic subjects and they increased significantly at 420 min after the bronchial challenges (P = 0.02, P = 0.03, respectively), while no significant changes were noted in group II subjects (P > 0.05).. These findings support the view that activated neutrophils may contribute to bronchoconstriction induced by TDI which may be associated with IL-8 release.

Topics: Adult; Asthma; Bronchi; Bronchoconstrictor Agents; Female; Humans; Interleukin-8; Male; Methacholine Chloride; Middle Aged; Neutrophil Activation; Occupational Diseases; Peroxidase; Sputum; Toluene 2,4-Diisocyanate

It has been demonstrated that consecutive samples of induced sputum may differ with respect to cellular composition. The aim of this study was to compare two sequential sputum samples in patients with chronic obstructive pulmonary disease (COPD) and asthma with different severity. Two sputum inductions were performed 30 min apart and processed separately in healthy subjects (n=11), patients with moderate to severe COPD (n=10), asthmatics treated with beta2-agonists alone (group 1, n=11), inhaled steroids (group 2, n=12) or systemic steroids (group 3, n=7). In healthy subjects and asthma group 2, percentages of neutrophils decreased significantly between the two sputum inductions but did not change in COPD and asthma group 3. Percentages of eosinophils did not change significantly in any group of patients. Concentrations of interleukin (IL)-8 decreased significantly in the control group and asthma groups 1 and 2 but not in asthma group 3 and the COPD group. These data demonstrate differences in sputum composition between two consecutive samples which were most pronounced in healthy subjects. Therefore, pooling of sputum samples may affect the results, particularly in healthy subjects, in contrast to subjects with more severe asthma or chronic obstructive pulmonary disease. These findings may be suggestive of differences in the distribution of inflammation along the airways between distinct airway diseases.

Topics: Administration, Inhalation; Administration, Oral; Adrenergic beta-Agonists; Adult; Aged; Albuterol; Asthma; Blood Proteins; Eosinophil Granule Proteins; Eosinophils; Female; Glucocorticoids; Humans; Interleukin-8; Lung Diseases, Obstructive; Male; Middle Aged; Prednisolone; Respiratory Function Tests; Ribonucleases; Sputum

Goblet cell (GC) hyperplasia and mucous plugging are common in patients with acute asthma. These patients also show neutrophil recruitment into the airways. Neutrophils contain elastase, a potent secretagogue in airways. Therefore, it was reasoned that neutrophil recruitment, by releasing elastase, could result in GC hypersecretion. When neutrophil chemoattractants were instilled in the airways of guinea-pigs, time-dependent neutrophil recruitment and GC degranulation occurred. An inhibitor of leukocyte infiltration (NPC15669) prevented both responses, implicating neutrophils. An inhibitor of neutrophil elastase (ICI 200,355) abolished GC degranulation, implicating elastase. Further studies implicate movement of elastase from cytoplasmic granules to the neutrophil surface, and they suggest a role for adhesion molecules on neutrophils and on GCs in neutrophil-dependent GC degranulation. Similarly, instillation of ovalbumin (OVA) into airways of OVA-sensitized guinea-pigs caused early recruitment of neutrophils and GC degranulation. GC degranulation was prevented by pretreatment with NPC15669 or ICI 200,355. These results implicate neutrophil release of elastase in allergen-induced hypersecretion. The results suggest a mechanism for the mucous plugging that occurs in acute asthma; prevention of neutrophil recruitment, prevention of neutrophil-GC adhesion, or inhibition of elastase activity could provide effective therapy for this serious pathophysiological abnormality.

Topics: Animals; Asthma; Cell Adhesion Molecules; Cell Degranulation; Goblet Cells; Guinea Pigs; Humans; Immunohistochemistry; In Vitro Techniques; Interleukin-8; Leukocyte Elastase; Male; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Ovalbumin; Swine

Evidence has now accumulated that heparin can significantly affect immune response including allergic inflammation. Cell migration is supposed to be very crucial in this process. Thus the aim of that study was to investigate whether low molecular weight heparin-nadroparine is chemoattractant for some inflammatory cells in asthmatics. Peripheral blood mononuclear cells (PBMC) and neutrophils from 15 asthmatics were obtained by gradient centrifugation. Chemotaxis was compared with that induced with known chemoattractants-fMLP and IL-8. We found that nadroparine caused significant and dose dependent chemotaxis of PBMC and comparable with influence of fMLP and IL-8. Nadroparine amplified chemotaxis of neutrophils but not significantly. Chemotaxis induced by fMLP and IL-8 was diminished when blood cells were incubated earlier with 50 mg of nadroparine.

Topics: Adult; Aged; Asthma; Chemotaxis; Female; Fibrinolytic Agents; Heparin, Low-Molecular-Weight; Humans; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Nadroparin; Neutrophils

To explore the relationship between interleukin-8 (IL-8) and airway hyper-responsiveness (AHR) of asthma in guinea pigs.. IL-8 at a dose of 0.5 microgram/kg or 5.0 micrograms/kg was administered intranasally to guinea pigs twice a week for 3 weeks. 24 hours after the last administration, airway responsiveness was measured as an overall index of airway response to the increasing concentrations of histamine inhaled (25, 50, 100 and 200 micrograms/ml), and the numbers of different inflammatory cells in bronchoalveolar lavage fluid (BALF) were counted.. The IL-8 treatment significantly enhanced airway responsiveness to histamine in a dose-dependent manner (P < 0.05 or P < 0.01) and induced a significant influx of neutrophils in BALF (P < 0.01), and a great number of neutrophils were seen within the airway wall, but not within the control animals treated with phosphate buffered saline (PBS).. The IL-8 could produce neutrophil inflammation of the airways and induce AHR, which suggested that IL-8 may play an important role in asthma, especially in the development of AHR.

Topics: Animals; Asthma; Bronchi; Bronchial Hyperreactivity; Guinea Pigs; Histamine; Interleukin-8; Lung; Male; Neutrophils; Random Allocation

To study the change of cytokines and eosinophil cationic protein (ECP) level in the sputum before and after glucocorticoid (GC) inhalation treatment so as to comprehend their effect on asthmatic and chronic obstructive pulmonary diseases (COPD) patients.. A method to induce sputum with inhaled hypertonic saline was used. The level of interleukin (IL)-5, IL-8 and ECP was measured with enzyme-linked immunosorbent assay method.. The concentration of ECP decreased from (500.3 +/- 49.6) microg/L to (59.8 +/- 10.9) microg/L, the percentage of eosinophils (Eos) dropped from (11.6 +/- 1.7) x 10(-2) to (4.1 +/- 0.7) x 10(-2) and there is significant difference in the concentration of IL-5 in the group of asthmatic patients after GC treatment. However, the concentration of IL-5 in the COPD patients did not show significant change after the same therapy.. Respiratory tract inflammation in asthma is related to Eos activation and increase in ECP and IL-5 excretion, while respiratory tract inflammation in COPD is related to neutrophil increase. These changes can be considered as the indicator of airway inflammation in asthma or COPD. Through regulating the quantity and function of the inflammatory cells and inhibiting the formation of cytokines to control the asthmatic airway inflammation, GC inhalation treatment will have better effect in treating asthmatic patients than COPD patients.

Topics: Administration, Inhalation; Adolescent; Adult; Asthma; Cytokines; Eosinophil Cationic Protein; Female; Glucocorticoids; Humans; Interleukin-5; Interleukin-8; Male; Middle Aged; Neutrophil Activation; Pulmonary Disease, Chronic Obstructive; Sputum

To investigate the adhesive mechanism of airway inflammation in the pathogenesis of asthma.. The guniea pigs were divided into 2 groups, the asthmatic animal group and the normal animal group. The following tests were performed: (a) The values of sICAM-1, sP-selectin, IL-8 and ECP both in plasma (or serum) and BALF were measured by ELISA. (b) The expressions of ICAM-1 and IL-8 on the bronchial epithelial cells and vascular endothelial cells were measured by immuno-histochemistry staining. (c) The lung functions (VT, Cdyn, R(L)) were tested.. (a) The values of sICAM-1, sP-selectin and ECP were elevated both in the plasma and BALF level of asthmatic group (P < 0.01). Additionally, IL-8 of BALF level of asthmatic group was elevated (P < 0.01), while IL-8 of serum level was lower than that of normal group (P < 0.01). (b) The expressions of ICAM-1, IL-8 on the bronchial epithelial and vascular endothelial cells in asthmatic animals were up-regulated (P < 0.01). (c) The lung functions of the asthmatic animals were impaired (V(T), C(dyn) P < 0.01; R(L) P < 0.05).. ALL the results above suggests that ICAM-1, IL-8, P-selectin and ECP are involved in the inflammative adhesive mechanism of asthma.

Topics: Animals; Asthma; Blood Proteins; Cell Adhesion; Eosinophil Granule Proteins; Guinea Pigs; Inflammation; Intercellular Adhesion Molecule-1; Interleukin-8; Lung; P-Selectin; Ribonucleases

Upregulation of adhesion molecule expression on endothelial cells (EC) and circulating leukocytes, by locally produced inflammatory mediators, may result in the enhanced infiltration of leukocytes into tissue, e.g. the airways of asthma patients. The present study investigates whether the expression of adhesion molecules on granulocytes and monocytes from asthma patients is affected by chemotactic factors, i.e. interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1). Flow cytometric analysis showed that the intrinsic expression of the various adhesion molecules on peripheral blood phagocytes from asthma patients was not different from that of healthy individuals. However, stimulation of monocytes with MCP-1 resulted only in upregulation of the expression of CD14 on monocytes from symptomatic asthma patients but not on monocytes from asymptomatic asthma patients and healthy individuals. Stimulation of granulocytes with IL-8 did not change the expression of the various beta 1- and beta 2-integrin molecules, such as VLA-4, LFA-1, CR3 and p150,95. Since earlier studies have shown that CD14 on monocytes mediates monocyte adhesion to activated vascular EC the present findings suggest that during the active phase of asthma upregulation of CD14 on monocytes by MCP-1 may lead to an increased adhesion of monocytes to vascular endothelium and their subsequent transendothelial migration into the tissue of the airways.

Topics: Adolescent; Adult; Asthma; CD18 Antigens; Cell Adhesion; Cell Adhesion Molecules; Cell Movement; Chemokine CCL2; Child; Endothelium, Vascular; Female; Granulocytes; Humans; In Vitro Techniques; Interleukin-8; Lipopolysaccharide Receptors; Male; Middle Aged; Monocytes; Up-Regulation

The adhesive interactions between phagocytes and endothelial cells (EC) can be modulated by inflammatory cytokines and chemotactic proteins which are released during an inflammatory response. The aim of the present study was to investigate first whether the adhesive properties of granulocytes and monocytes from asthma patients for vascular endothelial cells differ from those of phagocytes from healthy individuals. Furthermore, we studied whether the chemokines interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) can affect the binding of phagocytes to EC. No differences were observed in binding of phagocytes from asymptomatic or symptomatic asthma patients and from healthy individuals to non-stimulated or cytokine-stimulated EC. Incubation of granulocytes with IL-8 did not influence their adhesion to non-stimulated EC but inhibited the adhesion of granulocytes to IL-1-stimulated EC. Incubation of monocytes with MCP-1 did not affect their adhesion to non-stimulated or cytokine-stimulated EC. Our results indicate that adhesion of phagocytes to EC depends on the activation state of the endothelial cells but not on the origin of the phagocytes, since there were no differences in the adhesion of phagocytes from asthma patients and healthy individuals to non-stimulated or cytokine-stimulated EC.

Topics: Adolescent; Adult; Aged; Asthma; CD18 Antigens; Cell Adhesion; Cell Communication; Cells, Cultured; Chemokine CCL2; Endothelium, Vascular; Female; Granulocytes; Humans; In Vitro Techniques; Interleukin-1; Interleukin-8; Male; Middle Aged; Monocytes; Tetradecanoylphorbol Acetate

Topics: Anti-Bacterial Agents; Asthma; Clarithromycin; Eosinophils; Erythromycin; Fosfomycin; Humans; Hypersensitivity; Immunosuppressive Agents; In Vitro Techniques; Interleukin-8; Josamycin; Tacrolimus

The immuno-pathological mechanism for occupational asthma induced by grain dust (GD) remains to be clarified. There have been few reports suggesting the involvement of neutrophils inducing bronchoconstriction after inhalation of GD.. To further understand the role of neutrophil in the pathogenesis of GD-induced asthma.. We studied the phenotype of leucocytes of the bronchial mucosa in patients with GD-induced asthma. Bronchial biopsy specimens were obtained by fibreoptic bronchoscopy from six subjects with GD-induced asthma. Six allergic asthma patients sensitive to house dust mite were enrolled as controls. Bronchial biopsy specimens were examined by immunohistochemistry with a panel of monoclonal antibodies to tryptase-containing mast cell (AA1), activated eosinophil (EG2), pan T-lymphocyte (CD3) and neutrophil elastase (NE). Induced sputum was collected before and after the GD-bronchoprovocation test. The IL-8 level in the sputum was measured using ELISA.. There was a significant increase in the number of AA1+ and NE+ cells in bronchial mucosa of GD-induced asthma, compared with those of allergic asthma (P=0.01, P=0.01, respectively). No significant differences were observed in the number of EG2+ and CD3+ cells (P = 0.13, P=0.15, respectively). IL-8 was abundant in the sputum of all GD-induced asthma patients and significantly increased after the bronchial challenges compared with the baseline value (P = 0.03).. These findings support the view that neutrophil recruitment together with mast cells may contribute to the bronchoconstriction induced by GD. A possible involvement of IL-8 was suggested.

Topics: Antibodies, Monoclonal; Asthma; Biopsy; Bronchi; Dust; Edible Grain; Enzyme-Linked Immunosorbent Assay; Eosinophils; Humans; Immunohistochemistry; Interleukin-8; Mast Cells; Mucous Membrane; Neutrophils; Occupational Diseases; Receptor-CD3 Complex, Antigen, T-Cell; Respiratory Function Tests; Sputum

Previous studies have shown a significant association between confirmed diisocyanate-induced asthma (DOA) and in vitro production of diisocyanate antigen-stimulated histamine-releasing factors by PBMCs. Chemokines found in PBMC supernatants are known to express histamine-releasing factor activity.. PBMCs of diisocyanate-exposed workers were tested in vitro for diisocyanate antigen-specific enhancement of monocyte chemoattractant protein-1 (MCP-1), monocyte chemoattractant protein-3 (MCP-3), macrophage inflammatory protein-1alpha, RANTES, IL-8, and T-cell cytokines that could play a regulatory role in chemokine synthesis (IL-4, IL-5, IFN-gamma, and TNF-alpha.. Secretion of chemokines and cytokines was determined by quantitative immunochemical assays of PBMC supernatants. Synthesis of mRNA for beta-chemokines was determined by reverse transcription-polymerase chain reaction.. PBMCs of workers with DOA showed significantly enhanced secretion for MCP-1 compared with diisocyanate-exposed asymptomatic workers (P < .05). In vitro induction of antigen-stimulated MCP-1 mRNA synthesis in cultured PBMCs was demonstrated by reverse-transcription polymerase chain reaction. Quantitation of cytokines in supernatants showed increased mean production of IL-8 and TNF-alpha. IFN-gamma, IL-4, and IL-5 were not enhanced in subjects with DOA.. Antigen stimulation of MCP-1 and TNF-alpha suggest that diisocyanate-specific cellular immune reactions result in activation of macrophages, which may be important in the pathogenesis of DOA.

Topics: Adult; Antigens; Asthma; Biomarkers, Tumor; Chemokine CCL2; Chemokines; Cyanates; Female; Humans; Interleukin-8; Isocyanates; Leukocytes, Mononuclear; Lymphokines; Male; Middle Aged; Occupational Diseases; Toluene 2,4-Diisocyanate; Tumor Necrosis Factor-alpha; Tumor Protein, Translationally-Controlled 1

Recent studies have demonstrated that pulmonary surfactant protein (SP)-A plays a potential role in modifying inflammation and immune function. To see whether SP-A could modify IL-8 production and release by eosinophils stimulated with ionomycin, SP-A purified from surfactant recovered from patients with alveolar proteinosis was added to eosinophils isolated by the negative-selection method with immunomagnetic beads, and cultured for 24 h. The concentrations of IL-8 in the cell-free supernatants and cell lysates were then measured by ELISA. SP-A attenuated the production of IL-8 by eosinophils in a concentration-dependent manner. SP-A also attenuated the release of IL-8 from the eosinophils. The addition of SP-A antibody (PE10) reversed these effects of SP-A completely. These data suggest that SP-A may have the potential to modify allergic inflammation by inhibiting the release and production of IL-8 by eosinophils.

Topics: Antibodies, Monoclonal; Asthma; Dose-Response Relationship, Drug; Eosinophils; Humans; In Vitro Techniques; Inflammation; Interleukin-8; Ionomycin; Lung; Proteolipids; Pulmonary Surfactant-Associated Protein A; Pulmonary Surfactant-Associated Proteins; Pulmonary Surfactants

We examined the feasibility of using induced sputum to evaluate the airway inflammatory response to natural acute respiratory virus infections. We recruited eight asthmatics and nine healthy subjects on Day 4 of a cold. Viral infection was confirmed in six of the asthmatics (influenza A or B) and six of the healthy subjects (influenza A, rhinovirus, adenovirus, respiratory syncytial virus, and coronavirus). In the subjects with confirmed virus infection, five of the asthmatics had an objective exacerbation of asthma during the cold. Their sputum on Day 4 showed a high median total cell count of 19.7 x 10(6) cells/ml with a modest neutrophilia (58. 5%) and high levels of interleukin-8 (IL-8) (16,000 pg/ml), eosinophilic cationic protein (ECP) (1,880 microgram/L) and very high levels of fibrinogen (250 mg/L). In contrast, the proportion (1.3%) and absolute number of eosinophils was low. IL-2 levels were within the normal range, whereas IL-5 and interferon gamma were under the limit of detection of the assays. In the healthy subjects with a confirmed virus infection the sputum findings were qualitatively similar but significantly less prominent. Sputum IL-8 on Day 4 was strongly correlated with neutrophils (rs = 0.8, p < 0.001). This correlation was also significant when each group was analyzed separately. On Day 21 there was a fall in the absolute number of neutrophils and in ECP and fibrinogen levels in both groups. Similar results were found in the two asthmatic and three healthy subjects with a cold of comparable severity but in whom viral infection was not confirmed. We conclude that induced sputum examination can be used to study the effects of natural colds and influenza on the airways of the lungs. The results also suggest that natural colds, on Day 4, cause neutrophilic lower airway inflammation that is greater in asthmatics than in healthy subjects. The greater inflammatory response in asthmatics may be due to the changes associated with trivial eosinophilia or to the different viruses involved.

Topics: Acute Disease; Adenoviridae; Adult; Asthma; Blood Proteins; Common Cold; Coronavirus; Eosinophil Granule Proteins; Eosinophils; Feasibility Studies; Female; Fibrinogen; Humans; Inflammation; Inflammation Mediators; Influenza A virus; Influenza B virus; Influenza, Human; Interferon-gamma; Interleukin-2; Interleukin-5; Interleukin-8; Leukocyte Count; Male; Middle Aged; Neutrophils; Respiratory Syncytial Viruses; Rhinovirus; Ribonucleases; Sputum; Status Asthmaticus

Leukotriene B4 (LTB4), a potent chemokinetic mediator for neutrophils, is enhanced by interleukin-8 (IL-8) and may play a key role in the inflammatory response of asthma.. The aim of the present study was to investigate whether zileuton, a 5-lipoxygenase antagonist known to inhibit LTB4 production and recruitment of eosinophils/neutrophils in bronchoalveolar fluid, could affect the production of LTB4 and IL-8 by allergen-stimulated peripheral blood mononuclear cells in vitro from patients with asthma and/or allergic rhinitis.. Peripheral blood mononuclear cells were isolated using Ficoll-Hypaque density gradient from 14 subjects (2 with asthma, 11 with asthma and allergic rhinitis, and 1 with allergic rhinitis) and were stimulated by selected allergens (grass, tree, mite, and mold) in the absence or presence of 1 and 10 microM of zileuton. Supernatants were collected and assayed for LTB4 and IL-8 levels using RIA and ELISA, respectively.. Levels of LTB4 were significantly elevated in peripheral blood mononuclear cells stimulated with mold, grass, and tree compared with the unstimulated control group (P<.05). Levels of IL-8 were significantly elevated in all allergen-stimulated peripheral blood mononuclear cells, except mold, compared with the unstimulated control group (P<.05). Zileuton significantly reduced production of LTB4 by mold and tree-stimulated peripheral blood mononuclear cells. By contrast, no effect of zileuton on IL-8 production was observed in allergen-stimulated peripheral blood mononuclear cells.. The zileuton-induced attenuation of LTB4 production by allergen-stimulated peripheral blood mononuclear cells from patients with asthma and/or allergic rhinitis occurs independently from the allergen-stimulated IL-8 production.

Topics: Adult; Aged; Allergens; Asthma; Female; Humans; Hydroxyurea; Interleukin-8; Leukocytes, Mononuclear; Leukotriene B4; Lipoxygenase Inhibitors; Male; Middle Aged; Rhinitis, Allergic, Perennial; Rhinitis, Allergic, Seasonal

Asthma is considered a Th2-like disease, characterized by locally increased levels of interleukin (IL) 4. The bronchial epithelium plays an important role in the initiation and perpetuation of inflammatory reactions within the airways. However, little is known about the presence of IL-4 receptors on human bronchial epithelial cells, or the effects of IL-4 on these cells. In this report, definitive evidence of IL-4 receptor expression on human bronchial epithelial cells using several methods is presented. IL-4 receptor expression on human bronchial epithelial cells in vivo was demonstrated using in situ hybridization and immunohistochemistry. No difference in IL-4 receptor protein expression was observed between bronchial biopsies of healthy subjects compared to allergic asthmatics. Cultured human bronchial epithelial cells also expressed IL-4 receptor mRNA and protein (as determined by RT-PCR analysis and flow cytometry, respectively). IL-4 receptor protein expression by bronchial epithelial cells could be increased by stimulation with PMA+calcium ionophore, whereas IL-1beta and IL-6 decreased IL-4 receptor expression. A cyclic AMP analogue and IL-4 had no effect. Finally, it is shown that the IL-4 receptor is functionally active as IL-4 stimulates the release of IL-8, monocyte chemoattractant protein 1, and particularly IL-1 receptor antagonist by human bronchial epithelial cells. It is concluded that human bronchial epithelial cells express IL-4 receptors both in vivo and in vitro. Stimulation of human bronchial epithelial cells by IL-4 may result in the release of both pro- and anti-inflammatory mediators known to be upregulated in asthmatic airways.

Topics: Asthma; Bronchi; Bucladesine; Calcimycin; Cells, Cultured; Chemokine CCL2; Epithelial Cells; Gene Expression Regulation; Humans; In Situ Hybridization; Intercellular Adhesion Molecule-1; Interleukin 1 Receptor Antagonist Protein; Interleukin-4; Interleukin-8; Interleukins; Receptors, Interleukin-1; Receptors, Interleukin-4; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sialoglycoproteins; Tetradecanoylphorbol Acetate

Nitric oxide (NO) plays an important role as an inflammatory mediator in the airways. Though direct measurement of endogenous NO has been difficult in humans, we have recently found that measurement of NO derivatives in induced sputum may be useful for assessing airway inflammation in asthmatic patients.. This study was designed to determine the direct in vivo evidence of increased production of endogenous NO in patients with bronchial asthma and chronic obstructive pulmonary disease (COPD).. We have investigated simultaneous assessment of NO using two non-invasive methods, such as NO level in exhaled air and induced sputum, in these patients. We determined the concentration of stable end-products of NO (nitrite plus nitrate) in induced sputum and exhaled NO concentration using a chemiluminescence analyser in 10 normal controls, 10 asthmatic patients and 11 patients with COPD, and evaluated whether endogenous NO levels correlate with percentage of neutrophils and interleukin-8 (IL-8) level in induced sputum in patients with COPD.. We found significantly higher concentrations of exhaled NO in patients with bronchial asthma (25.1 [5.1] p.p.b.) than in patients with COPD (12.1 [1.9] p.p.b.) and normal controls (5.2 [1.4] p.p.b.). However, higher concentrations of NO derivatives in induced sputum were found in patients with bronchial asthma (1190 [106] micromol/L) and COPD (950 [105] micromol/L) than in normal controls (514 [30] micromol/L). In patients with asthma, but not in those with COPD, concentrations of NO derivatives in induced sputum were significantly correlated with concentrations of exhaled NO (r = 0.64, P < 0.05). Moreover, in patients with COPD, concentrations of NO derivatives in induced sputum were significantly correlated with percentage of neutrophils (r = 0.71, P < 0.05) and IL-8 level (r = 0.80, P < 0.01).. We conclude the increased production of endogenous NO in patients with asthma and COPD, and that NO derivatives in induced sputum are more valuable than exhaled NO in assessing airway inflammation in patients with COPD.

Topics: Adult; Aged; Asthma; Biomarkers; Eosinophils; Forced Expiratory Volume; Humans; Interleukin-8; Leukocyte Count; Lung Diseases, Obstructive; Male; Middle Aged; Neutrophils; Nitric Oxide; Sputum

Neutrophil infiltration is a major feature in the pathogenesis of the common cold, and respiratory viral infection is the major cause of asthma exacerbations. The factors regulating the neutrophil influx are unknown. Interleukin-8 (IL-8) is a potent neutrophil chemoattractant, which has been implicated in several inflammatory diseases. In this study, we investigated the presence of IL-8 chemokine in the nasal aspirates of asthmatic children (n = 12) in whom asthma was precipitated by proven viral infection. There were increased IL-8 levels in nasal aspirates from children during the virus-induced asthma exacerbations compared with samples from the same children when they had been asymptomatic for 2 wk (medians 863 and < 20 pg/ml, respectively, p < 0.01). Biological relevance was shown in that IL-8 levels correlate with increased nasal aspirate neutrophil myeloperoxidase levels and there was also a correlation between myeloperoxidase levels and upper respiratory symptom severity. Furthermore, we purified IL-8 from these samples, and demonstrated biological neutrophil chemotactic activity. These are the first in vivo data to suggest an important role for IL-8 in neutrophil influx in proven upper respiratory viral infection associated with asthma exacerbations. We suggest that IL-8 might provide a target for therapeutic intervention in virus-induced respiratory diseases.

Topics: Asthma; Chemotaxis, Leukocyte; Child; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Male; Nasal Mucosa; Neutrophil Activation; Neutrophils; Peroxidase; Respiratory Tract Infections; Virus Diseases

Although neutrophils have been implicated in bronchial asthma, the mechanism(s) which bring these cells into the airways is poorly understood.. To investigate the presence and identity of neutrophil chemotactic factors in bronchoalveolar lavage (BAL) fluid from atopic asthmatic subjects.. BAL fluid was obtained from 13 subjects (seven asthmatics and six normals), aged 19 to 60 yr, at bronchoscopy. Separation of neutrophil chemotactic activity (NCA) was achieved by FPLC cation exchange chromatography. Fractions were collected and assayed for chemotaxis in multiwell micro-chemotaxes chambers using polycarbonate filters, for the complement peptide C5a/C5a des Arg by radioimmunoassay (RIA) and for interleukin-8 (IL-8) by ELISA.. NCA was found in FPLC fractions of BAL samples in four out of seven asthmatics and each of these subjects had at least three similar peaks of NCA. The major peak of NCA was found to contain immunoreactive C5a/C5a des Arg and chemotaxis. In response to this NCA could be blocked by desensitization of the neutrophils with recombinant C5a. Purified serum derived C5a/C5a des Arg was found to have altered chromatographic properties when added to BAL fluid; this suggested that BAL fluid contained proteins which interacted with the C5a/C5a des Arg. Immunoreactive IL-8 (iIL-8) was also detected but its concentration or chemical form was insufficient to induce neutrophil chemotaxis.. This study demonstrates that bronchial asthmatic lavage fluid contains C5a/ C5a des/Arg and IL-8, together with other as yet unidentified factors which may contribute to neutrophil recruitment in this disease.

Topics: Adolescent; Adult; Asthma; Bronchoalveolar Lavage Fluid; Case-Control Studies; Chemotactic Factors; Complement C5a; Complement C5a, des-Arginine; Female; Humans; Interleukin-8; Male; Middle Aged; Neutrophils

We have tested the hypothesis that the expression of interleukin-8 (IL-8) is increased in bronchial tissue and circulating leukocytes of atopic asthmatics, indicating a role for this chemokine in asthma. The concentration of IL-8 in its free form and complexed with IgG or IgA was measured by ELISA in bronchial tissue, serum, and lysates of freshly isolated peripheral blood mononuclear cells and granulocytes from subjects with mild or severe asthma and nonatopic nonasthmatic subjects. Serum ECP was measured by fluorescent enzyme immunoassay. Free IL-8 was detected in the sera (n = 44) and bronchial tissue (n = 9) of all subjects with severe atopic asthma, but it was undetectable in normal subjects and subjects with mild atopic asthma, suggesting that free IL-8 is a marker of severe asthma. A positive correlation between free IL-8 and serum ECP levels found in severe disease suggests that IL-8 is associated with eosinophil activation. Complexes of IL-8 with IgA and IgG were detected in all serum and tissue samples. However, the levels of the IL-8-IgA complex were increased in the bronchial mucosa in asthma, and in blood were related to disease activity. Together, these results point to upregulation of IL-8 production in asthma and the induction of IL-8 binding immunoglobulins of the IgA class in the inflamed mucosa. We suggest a proinflammatory role for these complexes in lung tissue.

Topics: Asthma; Autoantibodies; Blood Proteins; Bronchi; Eosinophil Granule Proteins; Female; Humans; Inflammation Mediators; Interleukin-8; Leukocytes; Male; Mucous Membrane; Ribonucleases

Preliminary observations of the clinical efficacy of intravenous immunoglobulin in two patients with severe corticosteroid insensitive asthma are reported. In both patients treatment with intravenous immunoglobulin resulted in clinical improvement and enabled a significant reduction in the dose of prednisolone. In one of the patients fibreoptic bronchoscopy with endobronchial biopsies was performed and peripheral blood was analysed by flow cytometry before and after treatment. Immunohistological analysis of the biopsy samples after treatment showed a decrease in the number of all cell types, especially CD3+ T cells, CD4+ T cells, and activated CD25+ T lymphocytes, which was associated with a reduction in peripheral blood T cell activation. Intravenous immunoglobulin may be a valid option for the treatment of corticosteroid insensitive asthma. To elucidate the role and mode of action of intravenous immunoglobulin further studies in larger groups of patients are needed.

Topics: Adolescent; Asthma; B-Lymphocytes; Biomarkers; Blood Proteins; Budesonide; Chronic Disease; Combined Modality Therapy; Eosinophil Granule Proteins; Female; Glucocorticoids; Humans; Immunoglobulin E; Immunoglobulin G; Immunoglobulins, Intravenous; Inflammation Mediators; Interleukin-8; Lymphocyte Count; Prednisolone; Pregnenediones; Ribonucleases; T-Lymphocytes

To assess the characteristics of airway inflammation in patients with COPD.. We measured the sputum concentration of interleukin-8 (IL-8), a chemokine involved in the migration and activation of neutrophils and eosinophils. We also measured myeloperoxidase (MPO) as a parameter of neutrophil activity and eosinophil cationic protein (ECP) as a parameter of eosinophil activity. Spontaneous sputum samples were obtained from 33 patients with stable COPD and 30 patients with asthma. Induced sputum samples were obtained from 12 normal control subjects.. The sputum concentration of IL-8 was significantly higher in the patients with COPD than in the patients with asthma or in the control subjects (p<0.0001). Concentrations of MPO and ECP were significantly higher in the patients with COPD than in the control subjects but did not differ significantly between the patients with COPD and those with asthma. In the patients with COPD, the sputum concentration of IL-8 was significantly correlated with the concentration of MPO (r=0.55, p<0.001) and of ECP (r=0.53, p<0.01). The sputum concentration of IL-8 was negatively correlated with FEV1/FVC (r=-0.78, p<0.0001) in the COPD group.. Results suggest the activation of both neutrophils and eosinophils in the airways of patients with COPD. It appears that IL-8 plays a primary role in this activation. The sputum concentration of IL-8 appeared to be closely associated with the degree of airflow obstruction in patients with COPD and may serve as a marker in evaluating the severity of airway inflammation, which is a risk factor for COPD.

Topics: Adult; Aged; Asthma; Blood Proteins; Case-Control Studies; Eosinophil Granule Proteins; Eosinophils; Female; Humans; Inflammation Mediators; Interleukin-8; Lung Diseases, Obstructive; Lymphocyte Activation; Male; Middle Aged; Neutrophil Activation; Peroxidase; Respiratory Function Tests; Ribonucleases; Smoking; Sputum

The aim of this study was to determine the relative production of chemokines interleukin-8 (IL-8), regulated on activation, normal T-cell expressed and secreted (RANTES) and monocyte chemotactic protein-1 (MCP-1) by intrinsic and extrinsic asthmatics. Nine intrinsic asthmatics, 10 extrinsic asthmatics, five nonatopic and five atopic controls underwent bronchoalveolar lavage (BAL). Total BAL cells were cultured in the presence or absence of lipopolysaccharide. Chemokines were measured in BAL cell supernatants and in cell-free bronchoalveolar lavage fluid (BALF) by enzyme-linked immunoabsorbent assay (ELISA). BAL cell cytospins were stained immunohistochemically for chemokines. BAL cells from asthmatics produced more IL-8 than controls (statistically significant for extrinsic asthma). RANTES was elevated in the BAL cell supernatants of four out of nine intrinsic asthmatics as compared to nonatopic controls (not statistically significant). RANTES levels in the BAL cell supernatants of extrinsic asthmatics were all low. MCP-1 production by BAL cells was similar in all groups. Immunostaining of BAL cell cytospins showed the macrophage to be the predominant positive-staining cell type and correlated well with supernatant data. Measurement of chemokines in BALF showed significantly elevated IL-8 in intrinsic asthma compared to nonatopic controls, but no increase in extrinsic asthmatics relative to atopic control RANTES was elevated in three out of nine BALFs from intrinsic asthmatics compared with nonatopic controls (not statistically significant). MCP-1 was not elevated above control levels in BALF of either asthma group. These results suggest an up-regulation in the production of interleukin-8 and regulated on activation, normal T-cell expressed and secreted, but not monocyte chemotactic protein-1 (MCP-1), by macrophages in the bronchoalveolar lavage of asthmatic subjects. In addition, the data suggest that regulated on activation, normal T-cell, expressed and secreted, may be differentially produced by macrophages in atopic and nonatopic asthma.

Topics: Adult; Aged; Asthma; Bronchoalveolar Lavage Fluid; Cells, Cultured; Chemokine CCL2; Chemokine CCL5; Enzyme-Linked Immunosorbent Assay; Female; Humans; Hypersensitivity, Immediate; Immunohistochemistry; Interleukin-8; Lipopolysaccharides; Macrophages; Male; Middle Aged; Neutrophils; T-Lymphocytes

In studies of disease processes, increasing knowledge leads to an increased awareness of the complexity of the underlying mechanisms. The intense research activity in the chemokine field has made this acutely manifest. Numerous chemokines have been discovered through the use of (1) bioassay of in vitro cell culture supernatants and in vivo exudates from animal models of inflammation and (2) molecular biology techniques. Any one chemokine can often be produced by a number of different cell types and exert its effects on different target cells. This has been interpreted by some as implying a high degree of redundancy. Although this is understandable, in disease processes parallel and sequential mechanisms are possible, and potentially important therapeutic targets have emerged. There is compelling evidence from animal and clinical studies that eosinophils are important effector cells in asthma, but this relationship is as yet unproven in the human disease. Two possible targets to prevent eosinophil recruitment to the lung are IL-5 and its receptor, which are important in several aspects of eosinophil biology, and eotaxin and its receptor, CCR3. The eotaxin receptor is particularly attractive as a target as it is expressed in high numbers on eosinophils, but not other leukocytes, and appears to be the major detector of the eosinophil for eotaxin and other chemokines such as MCP-4. Eotaxin and CCR3 knockout mice are being developed, and animal models will continue to be invaluable when antagonists are available. In the shape of receptor antagonists, the chemokine field may yet provide the final proof of concept for the long-established eosinophil theory of asthma in humans.

Topics: Amino Acid Sequence; Animals; Asthma; Bronchoalveolar Lavage Fluid; Chemokine CCL11; Chemokine CCL2; Chemokine CCL5; Chemokines; Chemokines, CC; Chromatography, High Pressure Liquid; Cytokines; Disease Models, Animal; Endothelium, Vascular; Humans; Hypersensitivity; Interleukin-8; Leukocytes; Molecular Sequence Data; Ovalbumin

We report here the results of a multiphase project to assess the significance of airway responsiveness and airway injury in ozone (O3)* sensitivity. In Phase I, we measured the preexposure methacholine responsiveness of 66 normal subjects and then exposed these subjects to 0.2 ppm O3 for 4 hours with moderate exercise. Preexposure methacholine responsiveness was weakly correlated with O3-induced increases in specific airway resistance (sRaw) but not O3-induced declines in forced expiratory volume in one second (FEV1) or forced vital capacity (FVC). In addition, O3-induced lower respiratory symptoms were not well correlated with O3-induced changes in lung function. In Phase II, we exposed 23 normal subjects to O3, following an identical protocol to that of Phase I, and then performed bronchoscopy with proximal airway lavage (PAL), bronchoalveolar lavage (BAL), and bronchial biopsy at 18 hours after exposure. Ozone-induced increases in percentage of neutrophils and total protein concentration were observed in both bronchial fraction and BAL fluids; increased percentage of neutrophils also was observed in PAL fluid. These increases were correlated with O3-induced increases in sRaw, but not with O3-induced declines in FEV1 or FVC. Ozone also appeared to increase expression of intercellular adhesion molecule-1, an important mediator of neutrophil recruitment, in bronchial mucosa. In Phase III, we exposed a group of 19 asthmatic subjects to O3, following a protocol identical to that of Phase II. We then compared the lower respiratory symptom and lung function responses of the asthmatic subjects to those of the 81 normal subjects who participated in Phase I, Phase II, or both. The changes in the PAL and BAL fluids of the asthmatic subjects were compared with those of the normal subjects who participated in Phase II. Although both the asthmatic and nonasthmatic subjects showed significant O3-induced changes in lower respiratory symptoms, FEV1, FVC, and sRaw, no significant differences were found between the groups. For sRaw, however, a nonsignificant trend toward a greater O3-induced increase was noted for the asthmatic subjects. In contrast, the O3-induced increases in percentage of neutrophils and total protein concentration in BAL fluid were significantly greater for the asthmatic subjects than for the nonasthmatic subjects. These data suggest that although the lower respiratory symptom and lung function responses to O3 are not markedly greater in asthmatic su

Topics: Adolescent; Adult; Airway Resistance; Asthma; Biopsy; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Bronchoscopy; Data Interpretation, Statistical; Female; Forced Expiratory Volume; Humans; Immunohistochemistry; Inflammation; Interleukin-8; Lung; Male; Methacholine Chloride; Middle Aged; Ozone; Physical Exertion; Therapeutic Irrigation; Time Factors; Vital Capacity

To provide bases of comparison between the studies described in Parts I and II of this Research Report, concentrations of interleukin 6 (IL-6)*, interleukin 8 (IL-8), and alpha 2-macroglobulin (a2M) were measured in airway lavage fluids obtained in the Balmes study (Part I) and compared with the same measurements in the Frampton study (Part II). For healthy subjects in the Balmes study, IL-6 and a2M, but not IL-8, increased in association with ozone exposure. Statistical analyses suggested that effects of ozone on IL-8 levels observed in the first exposure and bronchoscopy may have carried over to the second exposure and bronchoscopy, which may have obscured an effect of ozone on IL-8 after the second exposure. For asthmatic subjects in the Balmes study, IL-6 and IL-8 increased in both bronchial and alveolar lavage fluid, but not in proximal airway lavage fluid. The mean interval between exposures was longer for asthmatic subjects than for healthy subjects, and no carryover effects were seen. When the Balmes and Frampton data were analyzed together, subject groups in the two studies (nonsmokers, smokers, and subjects without and with asthma) did not differ significantly in the response of cytokines to ozone exposure. The finding of possible carryover effects in one group suggests that subtle effects of ozone exposure, or bronchoscopy including proximal airway lavage and biopsy, or both, may persist for three weeks in some subjects.

Topics: Adolescent; Adult; alpha-Macroglobulins; Asthma; Biopsy; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Bronchoscopy; Data Interpretation, Statistical; Female; Humans; Inflammation; Inflammation Mediators; Interleukin-6; Interleukin-8; Lung; Male; Ozone; Smoking; Time Factors

To explore the relationship between interleukin-8 (IL-8) and airway hyperresponsiveness (AHR) of asthma.. IL-8 at a dose of 0.5 microgram/kg or 5 micrograms/kg was administered intranasally to guinea-pigs twice a week for 3 weeks. 24 hours after the last administration, airway responsiveness was measured as an overall index of airway response to increasing concentrations of inhaled histamine (25, 50, 100 and 200 micrograms/ml) and the numbers of different inflammatory cells in bronchoalveolar lavage fluid (BALF) was counted.. The IL-8 treatment significantly enhanced airway responsiveness to histamine in a dose-dependent manner (P < 0.05 or 0.01) and induced a significant influx of neutrophils in BALF (P < 0.01), and there are many neutrophils within airway wall but not control animals treated with phosphate buffered saline (PBS).. The IL-8 given into the airways can produce neutrophil inflammation of the airways and induce AHR, it may play an important role in asthma, especially in the development of AHR.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Guinea Pigs; Interleukin-8; Leukocyte Count; Male; Neutrophils

We have investigated whether IL-8 is present in airway secretions from patients with asthma and chronic obstructive pulmonary disease (COPD) to obtain information on its possible role in airway inflammation in obstructive airways disease. In the bronchoalveolar lavage fluid (BALF) from 11 clinically stable patients with asthma the levels of IL-8 were increased compared to 10 healthy subjects (median: controls 21.5 pg/ml, asthma 244 pg/ml: p < 0.005). In the patients with asthma the levels of IL-8 correlated with the percentage neutrophils in the BALF (r = 0.81; p < 0.001) and with a parameter of the permeability of the respiratory membrane, the quotient (alpha 2-macroglobulin in BALF)/(alpha 2-macroglobulin in serum) (r = 0.66; p < 0.025). In the sputum sol phase of 9 patients with symptomatic asthma the levels of IL-8 were lower than in 9 patients with COPD (asthma: 6.4 ng/ml; COPD: 16.3 ng/ml; p < 0.02) and significantly correlated with those of neutrophilic myeloperoxidase (MPO; r = 0.85; p < 0.005). The increased levels of IL-8 in the airway secretions from both patients with asthma and COPD may be markers of an ongoing inflammatory process, which is more pronounced in patients with COPD. In patients with asthma the strong correlation between the levels of IL-8 and the percentage neutrophils and/or the levels of MPO points to a role of IL-8 in the recruitment and activation of neutrophils in the airway lumen.

Topics: Adolescent; Adult; Aged; Asthma; Blood Proteins; Bronchoalveolar Lavage Fluid; Eosinophil Granule Proteins; Humans; Immunoglobulin A, Secretory; Inflammation Mediators; Interleukin-8; Lactoferrin; Lung Diseases, Obstructive; Middle Aged; Neutrophil Activation; Neutrophils; Permeability; Peroxidase; Ribonucleases; Sputum

Asthma and chronic obstructive pulmonary disease are characterized by chronic airway inflammation. Studies using bronchoalveolar lavage (BAL) have shown an increased proportion of eosinophils in the BAL fluid from asthmatics compared with that from normal subjects, whereas studies of chronic obstructive pulmonary disease (COPD) have shown increased numbers of neutrophils. Induced sputum allows sampling of respiratory tract secretions from patients and control subjects, providing a noninvasive method of studying airway secretions and allowing characterization of cells and measurement of soluble markers. We investigated whether induced sputum was a useful method of studying airway fluid from patients with moderate to severe COPD and whether it could be used to compare inflammation in this condition with that in asthma. An initial reproducibility study was undertaken. Sputum was induced twice in 13 patients with severe COPD at a 14-d interval. Total and differential cell counts were carried out and were found to be reproducible over this period. Sputum was then induced in 14 patients with COPD, 23 patients with asthma, 12 healthy cigarette smokers, and 16 normal nonsmoking control subjects. We found a significant increase in neutrophils and increased concentrations of tumor necrosis factor-alpha (TNF alpha) and interleukin-8 (IL-8) in the patients with COPD compared with the smoking and nonsmoking control subjects. Interleukin-8, but not TNF alpha, was significantly higher in the COPD group than in the asthmatic group. We conclude that the cytokines TNF alpha and IL-8 may be involved in the inflammation in COPD.

Topics: Adult; Aged; Asthma; Bronchoalveolar Lavage Fluid; Eosinophils; Female; Humans; Inflammation; Interleukin-8; Leukocyte Count; Lung Diseases, Obstructive; Male; Middle Aged; Neutrophils; Reproducibility of Results; Smoking; Sputum; Tumor Necrosis Factor-alpha

Chemokines are cytokines that induce chemotaxis of inflammatory cells. We studied the presence of chemokines in bronchoalveolar lavage fluid (BALF) obtained from nine allergic asthmatic patients and six nonsmoking normal individuals. The cells were pelleted, and ribonucleic acid (RNA) was extracted by using RNAzol B. BALF was assayed for monocyte chemoattractant protein-1 (MCP-1), regulated upon activation in normal T cells, expressed, probably secreted (RANTES), macrophage inflammatory protein-1alpha (MIP-1alpha) and interleukin-8 (IL-8) by enzyme-linked immunosorbent assay (ELISA). The levels of MCP-1, RANTES, and MIP-1alpha were significantly higher in the asthma patients than in the control subjects (p<0.04). The concentrations of RANTES and MCP-1 correlated with the lymphocyte count in the BAL specimens (r = 0.61 and 0.68, respectively). BALF showed eosinophil chemotactic activity in vitro that was blocked by anti-RANTES and anti-MCP-3 antibodies. The total cellular RNA was reverse-transcribed and the complementary deoxyribonucleic acid (cDNA) was amplified with the polymerase chain reaction (PCR) for MCP-1, MCP-3, RANTES, MIP-1alpha, IL-8, and beta-actin. We found that messenger ribonucleic acids (mRNAs) for MCP-1, MCP-3, RANTES, MIP-1alpha, and IL-8 were produced by BAL cells from most asthmatic and normal subjects. We conclude that chemokines are produced in the airways, and that an increased recovery of MCP-1, RANTES, and MIP-1alpha is observed in allergic asthmatic patients.

Topics: Adolescent; Adult; Asthma; Base Sequence; Bronchoalveolar Lavage Fluid; Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokine CCL5; Female; Humans; Interleukin-8; Lymphocyte Count; Macrophage Inflammatory Proteins; Male; Middle Aged; Molecular Sequence Data; Monokines

Topics: Alzheimer Disease; Animals; Asthma; Chemokine CCL11; Chemokine CCL2; Chemokines; Chemokines, CC; Chemotactic Factors, Eosinophil; Chemotaxis, Leukocyte; Cytokines; Humans; Inflammation; Interleukin-8; Monocytes; Neoplasms; Neutrophils; Respiratory Distress Syndrome

Interleukin 8 (IL-8) is a chemoattractant cytokine having a distinct target specificity for the neutrophil. However, it was also found to be active on primed eosinophils. IL-8 was measured by enzyme immunoassay in the bronchoalveolar lavage fluid (BALF) of 9 control subjects, 19 asthmatics and 36 chronic bronchitis patients. Its levels were correlated with neutrophil and eosinophil counts as well as levels of myeloperoxidase (MPO) and eosinophil cationic protein in the BALF. Asthmatic and chronic bronchitis patients had elevated levels of IL-8 in comparison with normal subjects. There was no correlation between the severity of asthma or chronic bronchitis and IL-8 levels. In asthmatic subjects there was a correlation between IL-8 and MPO levels (p < 0.0001, Spearman test). There was no significant correlation between eosinophils and IL-8. In chronic bronchitis there was a correlation between IL-8 and MPO levels as well as between IL-8 and neutrophil counts (p < 0.004 and p < 0.0152, respectively, Spearman test). IL-8 is involved in neutrophilic inflammation in asthma and chronic bronchitis, but differences in the IL-8 regulation of neutrophils were observed in both diseases.

Topics: Adolescent; Adult; Aged; Asthma; Bronchitis; Bronchoalveolar Lavage Fluid; Cell Count; Eosinophils; Humans; Interleukin-8; Middle Aged; Neutrophils; Peroxidase; Severity of Illness Index

We have investigated the profile of cellular recruitment into asthmatic airways after allergen and saline exposure and its relationship to interleukin-8 (IL-8) release. Fiberoptic bronchoscopy was used to instill allergen into the middle lobe while the right upper lobe received a sham saline challenge. Bronchoalveolar lavage (BAL) of both sites was performed either 4 or 24 h later. Neutrophil numbers in BAL fluid obtained 4 and 24 h after challenge were 17 and 48 times higher than prechallenge numbers (p < or = 0.001), but there was no statistically significant difference between the numbers of neutrophils at the two sites. In contrast, eosinophil numbers were increased by 6- and 20-fold, respectively, at 4 and 24 h at allergen-challenged as compared with saline-challenged sites (p < 0.005 and p < 0.02, respectively). Baseline concentrations of IL-8 in BAL fluid were undetectable in most cases. Four hours after allergen or saline exposure, BAL fluid IL-8 concentrations were: median, 200 pg/ml; range, 20 to 750 pg/ml and median, 123 pg/ml; range, < 20 to 800 pg/ml, respectively. These declined to 23 pg/ml (range, < 20 to 126 pg/ml) and 43 pg/ml (range < 20 to 130 pg/ml), respectively, 24 h after exposure. There was a significant correlation between neutrophil numbers and IL-8 concentrations 4 h after saline exposure. These findings indicate that neutrophil infiltration is a nonspecific response to the procedure of bronchoscopy and lavage, in contrast to eosinophil recruitment, which is an allergen-specific phenomenon, and it suggests that IL-8 release may be involved in neutrophil recruitment.

Topics: Adult; Albumins; Allergens; Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Bronchoscopy; Case-Control Studies; Cell Count; Enzyme-Linked Immunosorbent Assay; Eosinophils; Female; Humans; Interleukin-8; Male; Neutrophils; Time Factors

Interleukin-8 (IL-8) belongs to the family of chemotactic cytokines and has been shown to activate neutrophils in vitro and in vivo. In this study, we measured IL-8 concentration in the serum of patients with pulmonary emphysema or bronchial asthma. IL-8 concentration in serum of emphysema patients was significantly higher than in asthmatics; in emphysema patients it was significantly correlated with the smoking index and the annual decrease of FEV1.0. In asthmatics IL-8 concentration was below the level of detection, but was markedly increased during exacerbation of asthma. Our findings suggest that IL-8 may be one of the causal factors in these diseases.

Topics: Adolescent; Adult; Aged; Asthma; Chronic Disease; Female; Forced Expiratory Volume; Humans; Interleukin-8; Male; Middle Aged; Osmolar Concentration; Pulmonary Emphysema

Bronchial asthma is accompanied with inflammatory cell infiltration in the airway. Because kinin activity was detected in bronchoalveolar lavage fluid (BALF) from asthmatic patients, and because the concentrations of kallikrein and kinins in BALF increased after allergen challenge, we evaluated the potential that bradykinin (BK) might stimulate alveolar macrophages (AM) to release neutrophil, monocyte, and eosinophil chemotactic activity (NCA, MCA, and ECA). To test this hypothesis, bovine AM were isolated by bronchoalveolar lavage and cultured. The supernatant fluids of AM were tested for chemotactic activity by a blind well chamber technique. AM released NCA, MCA, and ECA in response to BK in a dose-and time-dependent manner (p < 0.01). The released activities were chemotactic by checkerboard analysis. Partial characterization and molecular sieve column chromatography revealed that these released activities were heterogeneous, suggesting predominant low-m.w. lipid soluble activity and weak high-m.w. peptide activity. Lipoxygenase inhibitors blocked the release of chemotactic activities (p < 0.01). The chemotactic activities were partly inhibited by leukotriene B4 (LTB4) and platelet-activating factor (PAF) receptor antagonists (p < 0.05, respectively). Immunoreactive LTB4 significantly increased in supernatant fluids in response to BK (p < 0.05), but PAF was detected only two out of six samples stimulated by BK. The receptor responsible for the release of LTB4 involved both BKB1 and BKB2 receptors. These data suggest that BK may stimulate AM and play a role in bronchial inflammation by recruiting inflammatory cells into the airway.

Topics: Animals; Asthma; Bradykinin; Bronchoalveolar Lavage Fluid; Cattle; Chemotactic Factors, Eosinophil; Cytokines; Dose-Response Relationship, Drug; Humans; Interleukin-8; Leukotriene B4; Lipoxygenase Inhibitors; Macrophages, Alveolar; Molecular Weight; Monocyte Chemoattractant Proteins; Platelet Activating Factor; Platelet Membrane Glycoproteins; Receptors, Cell Surface; Receptors, G-Protein-Coupled; Receptors, Leukotriene B4

Histamine-releasing factor consists of a group of cytokines that can cause basophils or mast cells to release histamine. However, the composition of histamine-releasing factor remains undefined.. This study was done to measure the concentrations, in plasma and mononuclear cell culture supernatants from children with asthma, of chemokines that are known to contribute to histamine-releasing factor activity.. Plasma and mononuclear cell culture supernatants were obtained from 25 children newly diagnosed with asthma, 25 good responders to immunotherapy, 23 poor responders, 25 patients with acute attacks, and 13 normal subjects. All the patient groups produced, spontaneously and after stimulation with phytohemagglutinin and mite allergen, greater amounts of monocyte chemotactic and activating factor, macrophage inflammatory protein-1 alpha, and RANTES (beta chemokines) and IL-8 and growth-related gene alpha (alpha chemokines) than did normal subjects. Successful immunotherapy resulted in decreased production, especially the spontaneous type, of beta chemokines that cause histamine release and in increased production of a chemokines that inhibit histamine release.. Abnormal chemokine production may contribute to the pathogenesis of bronchial asthma, and restoration of normal chemokine production may be used to explain, in part, the clinical efficacy of immunotherapy.

Topics: Adjuvants, Immunologic; Adolescent; Allergens; Animals; Asthma; Biomarkers, Tumor; Chemokine CCL2; Chemokine CCL4; Chemokine CCL5; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; Child; Female; Growth Substances; Humans; Immunosuppression Therapy; Immunotherapy; Intercellular Signaling Peptides and Proteins; Interleukin-8; Lymphokines; Macrophage Inflammatory Proteins; Male; Mites; Tumor Protein, Translationally-Controlled 1

The activity of neutrophil chemotactic factor (NCF) in serum of 40 patients with asthma on their acute attacks was measured with membrane filter method.. Of the 26 patients, 13 were administed salmeterol 50 micrograms bid by inhaler and another 13 took procaterol 50 micrograms bid orally for four weeks.. It was found that in patients with asthma NCF was 82 +/- 17 cell/10 HP, which was significantly higher than that in normal subjects (33 +/- 5 cell/10 HP, P < 0.01). After treatment with salmeterol the decrease in NCF activity was 26 cell/10 HP (P < 0.01), while the decrease in another group treated with procaterol was 8 cell/10 HP (P > 0.05). The difference between the two treated groups was significant (P < 0.01). In salmeterol treated group the decrease in NCF activity was more remarkable than the improvement in lung function.. It was considered that the result may be related to the anti-inflammatory effect of salmeterol.

Topics: Adrenergic beta-Agonists; Adult; Albuterol; Asthma; Female; Humans; Interleukin-8; Male; Middle Aged; Procaterol; Respiratory Function Tests; Salmeterol Xinafoate

The importance of cytokines in the asthmatic inflammatory response is becoming apparent. The aim of this study was to determine whether the non-invasive method of induced sputum combined with the polymerase chain reaction would allow the detection of messenger RNA (mRNA) encoding a range of cytokines on a qualitative basis.. Four groups were studied comprising 10 normal subjects, six atopic, 10 mild and five moderately severe asthmatic subjects. Sputum was induced by the inhalation of nebulised 3.5% saline and total RNA extracted from the expectorated cells. Expression of cytokine message within induced sputum was examined by reverse transcription and the polymerase chain reaction (PCR) using primers specific for a range of cytokines (IL-1 alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, RANTES, TNF alpha, IFN alpha, IFN gamma). Presence or absence of the signal was determined at 35 and 70 cycles of PCR by electrophoretic size fractionation on ethidium bromide stained agarose gels.. Cytokine message was detectable in sputum by this method. All samples showed a positive result for actin control. Analysis of signal for the cytokines in all subjects showed that, at 70 cycles, IL-1, IL-5, IL-8, and TNF alpha were detected in more subjects than would be expected by chance. IL-5 mRNA was detected in more of the asthmatic patients (moderate 80%, mild 40%) than in the atopic subjects (33%), who in turn showed expression of this cytokine in more individuals than nonatopic subjects (10%).. The combination of sputum induction and PCR appears to be a useful, non-invasive tool to explore the chronic inflammation of asthma and possibly other lung disorders. It should enable differences between normal and asthmatic subjects to be identified for future confirmation by quantitative techniques.

Topics: Adult; Asthma; Base Sequence; Cytokines; Female; Humans; Hypersensitivity, Immediate; Interleukin-1; Interleukin-5; Interleukin-8; Male; Middle Aged; Molecular Sequence Data; Polymerase Chain Reaction; RNA, Messenger; Sputum; Tumor Necrosis Factor-alpha

The effects of selective phosphodiesterase (PDE) isoenzyme inhibitors on eosinophil airway infiltration induced by intratracheal administration of recombinant human cytokines were investigated in the guinea pig. Recombinant human IL-5 and IL-8 elicited a concentration-dependent increase in the number of eosinophils in the bronchoalveolar lavage (BAL) fluid. In contrast, no effect was observed after intratracheal injection of recombinant human IL-3 or recombinant human RANTES. Pretreatment with the PDE IV inhibitors rolipram or Ro 20-1724 or the nonselective PDE inhibitor theophylline 1 h before intratracheal injection of IL-5 significantly reduced the number of eosinophils in the BAL fluid at 48 h. In contrast, the selective PDE III inhibitors milrinone and SK&F 94-836 and the PDE I/V inhibitor zaprinast did not inhibit the airway eosinophil infiltration induced by IL-5. Betamethasone also significantly inhibited the IL-5-induced eosinophil infiltration in BAL fluid. Administration of rolipram or betamethasone 1 h before IL-8 significantly reduced airway eosinophil infiltration. Because the selective PDE IV inhibitors markedly inhibited eosinophil infiltration in guinea pig airways induced by cytokines, it is suggested that PDE IV inhibitors have antiinflammatory effects in the airways and may be useful in the treatment of asthma.

Topics: Animals; Asthma; Betamethasone; Bronchoalveolar Lavage Fluid; Chemokine CCL5; Cytokines; Eosinophils; Guinea Pigs; Interleukin-3; Interleukin-5; Interleukin-8; Lymphokines; Male; Phosphodiesterase Inhibitors; Premedication; Recombinant Proteins; Time Factors; Trachea

Inflammation can be demonstrated in the airway mucosa of asthmatics, even in the absence of overt symptoms, but the pathogenesis of this chronic inflammation is incompletely defined. It has been suggested that inflammatory cytokines produced by epithelium may play important roles in this process. Therefore, we measured the cytokines interleukin-8 (IL-8), IL-6, and granulocyte-macrophage colony-stimulating factor (GM-CSF) in nasal lavage fluids from school-age children who were (1) "normal" (nonallergic/nonasthmatic), (2) allergic to house-dust mite antigen but nonasthmatic (no history of wheezing), or (3) allergic and asthmatic (history of > or = 10 wheezing episodes). Children underwent a single nasal lavage procedure while asymptomatic and on no anti-inflammatory medications or anti-histamines. In addition to cytokine concentrations, cell counts, differentials, albumin, histamine, and eosinophil cationic protein (ECP) concentrations were determined in nasal lavage fluids. Significant increases in IL-8 and ECP were observed in asthmatics compared with both normals and allergic nonasthmatics. Overall, IL-8 in nasal lavage fluids correlated significantly with ECP. Allergic nonasthmatics did not have significant increases in cytokines or other mediators compared with normal subjects. Concentrations of IL-6 did not differ significantly among the three groups, and GM-CSF was undetectable in all samples tested. We conclude that increased IL-8 production and eosinophil activation are characteristic of the airways of asthmatic children when asymptomatic, and we speculate that IL-8 plays a role in the maintenance of airway inflammation in asthma.

Topics: Adolescent; Animals; Asthma; Blood Proteins; Case-Control Studies; Child; Cytokines; Dust; Enzyme-Linked Immunosorbent Assay; Eosinophil Granule Proteins; Female; Humans; Inflammation Mediators; Interleukin-8; Longitudinal Studies; Male; Mites; Nasal Lavage Fluid; Respiratory Hypersensitivity; Ribonucleases

Adenosine potentiates mast cell activation, but the receptor type and molecular mechanisms involved have not been defined. We, therefore, investigated the effects of adenosine on the human mast cell line HMC-1. Both the A2a selective agonist CGS21680 and the A2a/A2b nonselective agonist 5'-N-ethylcarboxamidoadenosine (NECA) increased cAMP, but NECA was fourfold more efficacious and had a Hill coefficient of 0.55, suggesting the presence of both A2a and A2b receptors. NECA 10 microM evoked IL-8 release from HMC-1, but CGS21680 10 microM had no effect. In separate studies we found that enprofylline, an antiasthmatic previously thought to lack adenosine antagonistic properties, is as effective as theophylline as an antagonist of A2b receptors at concentrations achieved clinically. Both theophylline and enprofylline 300 micro completely blocked the release of IL-8 by NECA. NECA, but not CGS21680, increases inositol phosphate formation and intracellular calcium mobilization through a cholera and pertussis toxin-insensitive mechanism. In conclusion, both A2a and A2b receptors are present in HMC-1 cells and are coupled to adenylate cyclase. In addition, A2b receptors are coupled to phospholipase C and evoke IL-8 release. This effect is blocked by theophylline and enprofylline, raising the possibility that this mechanism contributes to their antiasthmatic effects.

Topics: Adenosine; Adenosine-5'-(N-ethylcarboxamide); Anti-Asthmatic Agents; Asthma; Calcium; Cyclic AMP; Humans; Inositol Phosphates; Interleukin-8; Mast Cells; Receptors, Purinergic P1; Tumor Cells, Cultured; Xanthines

Accumulation of CD4+ interleukin (IL)-2R+ lymphocytes in the airways of asthmatics is generally attributed to the presence of chemoattractant cytokines. The precise mechanism for the initiation of the earliest CD4+ lymphocyte infiltration and activation is unknown. In this study, we describe for the first time the presence of lymphocyte chemoattractant activity in the bronchoalveolar lavage (BAL) fluid obtained from asthmatics 6 h after antigen challenge. The majority of the chemoattractant activity at this early time point is represented by IL-16 (lymphocyte chemoattractant factor), a CD4+ cell-specific chemoattractant and growth factor. In addition to IL-16, macrophage inflammatory protein 1 alpha (MIP1 alpha) chemotactic bioactivity was detected in significant levels. While IL-16, MIP1 alpha, and IL-8 were all identified by enzyme-linked immunosorbent assay, the great majority of the lymphocyte chemoattractant activity in the BAL fluid after antigen challenge is attributable to IL-16 and MIP1 alpha. There were no detectable levels of IL-16 nor MIP1 alpha in BAL fluid of antigen-challenged normal subjects nor atopic nonasthmatics nor in saline-challenged lobes from the asthmatics. The identification of multiple lymphocyte chemoattractants early after antigen challenge suggests a complex cellular, as well as chemoattractant cytokine, profile in initiating the CD4+ T cell-mediated inflammatory process that is specific for the atopic asthmatic phenotype.

Topics: Adult; Antigens; Asthma; Bronchoalveolar Lavage Fluid; Chemokine CCL5; Chemotaxis; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Lymphocytes; Male; Middle Aged; Receptors, Chemokine; Receptors, Immunologic; Succinimides; Time Factors

Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin (IL)-5 or IL-8 have been suggested to play an important role in the pathogenesis of eosinophilic airway inflammation in bronchial asthma or neutrophilic airway inflammation in chronic bronchitis, respectively, However, GM-CSF and IL-8 have biological activities to either eosinophils or neutrophils.. To investigate the contribution of these cytokines to airway inflammation, we compared the cellular differential and immunolocalization of GM-CSF, IL-5 and IL-8 in sputum cells from patients with bronchial asthma and chronic bronchitis.. Cytospins of sputum cells from 12 patients with bronchial asthma and 12 with chronic bronchitis were subjected to cellular differential counting and immuno-cytochemistry with anti-human GM-CSF, IL-5 and IL-8 antibody.. The predominant cells in bronchial asthma were eosinophils and lymphocytes, while those in chronic bronchitis were neutrophils. All cytokines examined were detected in either bronchial asthma or chronic bronchitis, although the percentage of GM-CSF and Il-5 positive cells in bronchial asthma (53.4 +/- 6.0 [mean +/- SEM]% and 9.7 +/- 2.8%, respectively) was significantly higher than that in chronic bronchitis (11.4 +/- 2.5%; P < 0.001 and 1.7 +/- 0.3%; P < 0.007, respectively). In contrast, the percentage of IL-8 positive cells in chronic bronchitis (23.8 +/- 7.0%) was significantly higher than that in bronchial asthma (7.& +/- 1.9%; P < 0.04). The cells positive for IL-5 were lymphocytes in bronchial asthma and chronic bronchitis. The cells positive for GM-CSF in bronchial asthma were predominantly eosinophils, while those in chronic bronchitis were monocytes/macrophages and neutrophils. In contrast, neutrophils are mainly positive for IL-8 in chronic bronchitis, while monocytes/macrophages and bronchial epithelial cells are positive for IL-8 in bronchial asthma.. The immunochemical comparison of GM-CSF and IL-8 localization in sputum cells between bronchial asthma/chronic bronchitis suggests the differential regulation and roles of these cytokines in eosinophilic vs neutrophilic airway inflammation, resulting in the development of different types of airway inflammation.

Topics: Adult; Aged; Aged, 80 and over; Asthma; Bronchitis; Child; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-5; Interleukin-8; Male; Middle Aged; Sputum

Interleukin-8 (IL-8) has been shown to be a chemotactic factor for neutrophils, T-lymphocytes and eosinophils, but it is unknown whether the IL-8-induced inflammatory cell accumulation into the airways can cause the bronchial hyperresponsiveness (BHR) characteristic of asthma. IL-8 at a dose of 0.5 or 5 micrograms/kg was administered intranasally to guinea-pigs twice a week for 3 weeks. One day after the last administration, animals were anesthetized and artificially ventilated through tracheal cannula and lateral pressure at the cannula (Pao) was measured as an overall index of airway responses to increasing concentrations of inhaled histamine (25, 50, 100, and 200 micrograms/ml). The IL-8 treatment significantly enhanced bronchial responsiveness to histamine in a dose-dependent manner (ANOVA P < 0.01). The provocative concentration of histamine causing a 100% increase in Pao (PC100) at a dose of 0.5 and 5 micrograms/kg of IL-8 was 68.1 (GSEM 1.12) and 35.6 (GSEM 1.25) micrograms/ml, respectively. The latter was significantly (P < 0.01) lower than that in control animals treated with PBS (93.3 [GSEM, 1.14] micrograms/ml). The IL-8 treatment also induced a significant influx of neutrophils, but not eosinophils, in bronchoalveolar lavage (BAL) fluid (18.3 +/- 8.8 and 30.6 +/- 8.3% in animals treated with 0.5 and 5 micrograms/kg, respectively, of IL-8 vs 3.6 +/- 0.7% in phosphate buffered saline-(PBS)-treated animals). Furthermore, we examined the effect of the thromboxane receptor antagonist S-1452 (0.01 or 0.1 mg/kg, i.p. 24 and 1 h before anesthesia) on this IL-8 induced BHR. S-1452 significantly inhibited the BHR dose-dependently (ANOVA P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Administration, Intranasal; Animals; Asthma; Bridged Bicyclo Compounds; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Fatty Acids, Monounsaturated; Guinea Pigs; Interleukin-8; Male; Neutrophils; Receptors, Prostaglandin; Thromboxane A2

Eosinophils possess the capacity to synthesize various cytokines. We demonstrate that IL-8 mRNA and protein are constitutively expressed by freshly isolated resting human eosinophils. Most of the patients with bronchial asthma or atopic dermatitis show evidence for up-regulated IL-8 protein expression in eosinophils but not in neutrophils, suggesting that an eosinophil-specific cytokine may act in these patients. To investigate whether the intracellular IL-8 can be released, eosinophils were stimulated by different cytokines and platelet-activating factor. Priming with granulocyte-macrophage CSF and a subsequent 25-min stimulation with RANTES or platelet-activating factor resulted in release of IL-8 from highly purified human eosinophils in vitro. As the eosinophil is the predominant cell in asthmatic inflammation, we determined IL-8 concentrations in bronchoalveolar lavage fluids from normal individuals and asthmatic patients. Bronchoalveolar lavage fluids from patients with bronchial asthma consistently demonstrated high IL-8 concentration compared with the controls. This suggests that IL-8 is released in vivo by inflammatory bronchial cells in asthma.

Topics: Asthma; Base Sequence; Bronchoalveolar Lavage Fluid; Eosinophils; Flow Cytometry; Humans; Immunoblotting; Immunohistochemistry; Interleukin-8; Molecular Sequence Data; Neutrophils; Precipitin Tests; RNA, Messenger

Although glucocorticosteroids are a very effective treatment for asthma and other chronic inflammatory diseases, a small proportion of patients are resistant to their therapeutic effects. The molecular mechanism for this steroid resistance is unclear. Steroid resistance cannot be explained by pharmacokinetic mechanisms, by a defect in the binding of steroids to glucocorticoid receptors, nor by defective nuclear translocation of this receptor, thereby suggesting that the molecular abnormality lies distal to nuclear translocation. We examined the ability of nuclear translocated glucocorticoid receptors to bind to their DNA binding sites (GRE) using electrophoretic mobility shift assays in PBMC from patients with steroid-sensitive and steroid-resistant asthma. The binding of the glucocorticoid receptor to DNA in these patients was also studied using Scatchard analysis. Dexamethasone induced a significant rapid and sustained twofold increase in GRE binding in PBMCs from steroid-sensitive asthmatic patients and nonasthmatic individuals, but this was markedly reduced in steroid-resistant asthmatic patients. Scatchard analysis of glucocorticoid receptor-GRE binding showed no change in binding affinity but did show a reduced number of receptors available for DNA binding in the steroid-resistant patients. These results suggest that the ability of the glucocorticoid receptor to bind to GRE is impaired in steroid-resistant patients because of a reduced number of receptors available for binding to DNA.

Topics: Adult; Asthma; Base Sequence; Cells, Cultured; DNA-Binding Proteins; Drug Resistance; Electrophoresis, Polyacrylamide Gel; Female; Glucocorticoids; Humans; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Molecular Sequence Data; Polymerase Chain Reaction; Receptors, Glucocorticoid; RNA, Messenger

Previous work has demonstrated an increase in the production of granulocyte-macrophage colony-stimulating factor (GM-CSF) by monocytes derived from asthmatic individuals. We have suggested that monocytes and macrophages enhance airways inflammation by augmented cytokine production. We tested this hypothesis by measuring the production of GM-CSF and macrophage-derived cytokines, namely interleukin-1 beta (IL-1 beta), tumour necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8), from unstimulated and lipopolysaccharide (LPS)-stimulated peripheral blood monocytes and alveolar macrophages in 31 asthmatic and 11 normal, control subjects. The basal production of GM-CSF was four fold higher in the monocytes of asthmatic individuals, but there was no significant difference in the basal production of TNF-alpha, IL-1 beta and IL-8. After stimulation with LPS, asthmatic monocytes produced twofold more GM-CSF and fourfold more IL-1 beta than the monocytes from control subjects. Unstimulated macrophages from asthmatic subjects produced significantly less GM-CSF and TNF-alpha than macrophages from controls, and there was no difference in either IL-1 beta or IL-8 production. When stimulated by LPS, macrophages from asthmatic subjects produced twofold more GM-CSF, threefold more TNF-alpha and fourfold more IL-8. The levels of IL-8 produced by both monocytes and macrophages were at least 20 fold higher than those of the other cytokines measured. There is selectivity in the upregulation of cytokine production by monocytes and macrophages in asthma.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Asthma; Bronchoalveolar Lavage Fluid; Cells, Cultured; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-1; Interleukin-8; Lipopolysaccharides; Macrophages, Alveolar; Male; Monocytes; Tumor Necrosis Factor-alpha

Topics: Asthma; Cell Movement; Chemotaxis, Leukocyte; Eosinophils; Humans; Interleukin-8; Lung

Plasma interleukin-8 (IL-8) concentrations were measured in patients with atopic dermatitis. Plasma IL-8 was not detected in 25 controls (0/25), in allergic rhinitis (0/20), or in bronchial asthma during remission (0/13), while low concentrations of IL-8 were detectable in a few patients with urticaria (1/19), contact dermatitis (4/17), and bronchial asthma at the time of attack (6/16). In contrast, IL-8 was detectable in most cases of atopic dermatitis (41/52). Moreover, IL-8 concentrations were significantly higher in severe than in mild or moderate atopic dermatitis. IL-8 concentrations decreased as atopic dermatitis was improved by treatment, and IgE production in vitro was also decreased while serum IgE concentrations remained unchanged. IL-8 measurement may be a useful tool for the study of the pathogenesis and clinical course of atopic dermatitis.

Topics: Adolescent; Asthma; Cells, Cultured; Child; Child, Preschool; Dermatitis, Atopic; Dermatitis, Contact; Female; Humans; Immunoglobulin E; Infant; Interleukin-1; Interleukin-6; Interleukin-8; Male; Rhinitis; Urticaria

The objective of this study was to determine whether exposure to ozone causes inflammatory or functional changes in the upper or lower airways of asthmatic and nonasthmatic individuals. Ten asthmatic and eight nonasthmatic subjects were exposed to clean air, 120 ppb ozone, or 240 ppb ozone for 90 min during intermittent moderate exercise using a head dome exposure system. Pulmonary function tests, posterior rhinomanometry, and nasal lavage were performed before and after exposure. Leukocyte counts and chemotactic factors leukotriene B4 (LTB4), platelet-activating factor (PAF), and interleukin-8 (IL-8) were analyzed from nasal lavage fluid. In subjects with asthma, a significant increase (p < 0.05) in the number of white blood cells in lavage fluid was detected both immediately and 24 h after exposure to 240 ppb ozone, as was a significant increase in epithelial cells immediately after exposure (p < 0.05). No significant cellular changes were seen in nonasthmatic subjects. A significant correlation was observed between IL-8 and white blood cells counts after exposure to 240 ppb ozone (r = 0.76) in asthmatic subjects. No significant changes in pulmonary or nasal function or biochemical mediators were found in either the asthmatic or nonasthmatic subjects. These data indicate that asthmatic individuals are more sensitive to the acute inflammatory effects of ozone than nonasthmatic individuals.

Topics: Adolescent; Adult; Asthma; Female; Histamine; Humans; Inflammation; Interleukin-8; Leukocyte Count; Leukotriene B4; Male; Nasal Lavage Fluid; Ozone; Platelet Activating Factor; Respiratory Mechanics; Respiratory System

T lymphocytes may orchestrate the inflammatory response in atopic asthma, but the mechanisms that promote T-cell accumulation in asthmatic airways are still unclear. In this study, we tested the hypothesis that bronchial epithelial cells of patients with atopic asthma release chemoattractant factors for T lymphocytes.. Sixteen patients with atopic asthma and eight healthy control subjects were selected for this study. Bronchial epithelial cells were isolated from biopsy specimens obtained by means of bronchoscopy and cultured for 48 hours in serum- and hormone-free medium, with or without 10(-6) mol/L histamine.. Only the supernatants of cells from donors with asthma showed chemotactic activity for T lymphocytes, and this was significantly increased (p < 0.025) by exposure to histamine. Chemotactic activity was in part mediated by interleukin-8 (IL-8), because an antibody against human IL-8 significantly reduced it (p < 0.05) and the cell supernatants contained appreciable amounts of immunoreactive IL-8 (0.89 +/- 0.39 ng/ml). Both the residual chemotactic activity of unstimulated epithelial cells and the increased activity caused by histamine were mediated by a single protease-sensitive substance with an apparent molecular weight of 56,000 d and an estimated isoelectric point of 8.8 to 9.1. The partially purified chemoattractant specifically enhanced the migration of CD4+ T lymphocytes, and its activity was inhibited by the univalent Fab fragment of a monoclonal antibody against CD4.. These results extend our previous observations, indicating an important effector role of bronchial epithelium in asthma.

Topics: Adult; Asthma; Bronchi; Cells, Cultured; Chemotactic Factors; Chemotaxis, Leukocyte; Epithelium; Female; Histamine; Humans; Interleukin-8; Male; T-Lymphocytes

The cytokine granulocyte-macrophage colony-stimulating factors, interleukin-3 and interleukin-5, are important modulators of eosinophilia and eosinophil function. In particular, eosinophil chemotaxis is very sensitive to cytokine priming.. We evaluated chemotactic responses of eosinophils from patients with allergic asthma. These cells exhibited a primed phenotype as deduced from enhanced responses toward formyl-methionyl-leucyl-phenylalanine and platelet-activating factor and a decreased responsiveness toward granulocyte-macrophage colony-stimulating factor. Bronchoprovocation of patients with allergic asthma with allergen was performed as a possible means to enhance in vivo priming.. Indeed, eosinophils isolated 3 hours after allergen challenge exhibited a more pronounced primed phenotype, which was reflected by an induction of responsiveness towards interleukin-8. Eosinophil responses induced by platelet-activating factor, formyl-methionyl-leucyl-phenylalanine, complement fragment C5a, interleukin-3, interleukin-5, and granulocyte-macrophage colony-stimulating factor were not significantly altered after allergen challenge.. These data provide further evidence that eosinophils are already primed in the peripheral blood of individuals with allergic asthma. This is most likely due to the presence of circulating cytokines in the peripheral blood of those individuals. This in vivo priming results in selective upregulation and downregulation of responses toward various chemotaxins, which may be released in the lungs during allergic inflammation.

Topics: Adult; Asthma; Bronchial Provocation Tests; Chemotaxis, Leukocyte; Complement C5a; Eosinophils; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Male; Middle Aged; N-Formylmethionine Leucyl-Phenylalanine; Platelet Activating Factor; Up-Regulation

To evaluate an effect of Intal on the late post-provocation bronchospasm in 30 patients with atopic asthma (with early bronchospasm) the treatment was accompanied by the assessment of NCF activity (as a marker of mastocytes degranulation. It was shown that after therapy with Intal exerted an effect on the NCF activity not only in the early but also late bronchospasm in comparison with its activity in patients after exercise test but prior to therapy--9/22 and 6/29. The authors share an opinion tha the presence of NCF in part of patients with the late postexercise bronchospasm (treated with Intal) may indicate that its origin is different (not from mastocytes).

Topics: Asthma; Bronchial Provocation Tests; Cromolyn Sodium; Exercise Test; Humans; Interleukin-8

To assess the incidence of the late reaction of the bronchi in patients with atopic asthma the following methods of the provocation with atopic asthma the following methods of the provocation were applied: a) exercise test, b) a 3-minute ventilation with cold air and c) inhalation of the distilled water Disturbances of ventilation were assessed spirographically. An emphasis was also on the activity of NCF in the blood serum following the applied provocation tests. It was found that the late bronchospasm followed the exercise test in about half of the examined patients (48%). It was less frequent after hyperventilation with cold air (33%) and inhalation of the distilled water (38%). Bronchospasm was accompanied by the increase in NCF activity in the blood serum. The late bronchospasm after provocation tests in asthmatics is a real fact, not depending on the accidental ventilation disorders.

Topics: Adult; Asthma; Bronchial Provocation Tests; Bronchial Spasm; Female; Humans; Interleukin-8; Male

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Asthma; Bronchi; Cells, Cultured; Cytokines; Epithelium; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Hydrocortisone; Interleukin-6; Interleukin-8; Male; Nedocromil; Quinolones

To clarify whether thromboxane A2 (TXA2) is involved in type III and IV allergy, or so called "cell-mediated allergy", we studied the effect of a specific TXA2 synthetase inhibitor, sodium ozagrel (OKY-046) on peripheral blood mononuclear cells and neutrophils in adult intractable asthmatics. The results revealed, firstly, that lymphocyte blastogenesis and interleukin-2 (IL-2) production from peripheral blood mononuclear cells stimulated by PHA and Candida antigen in intractable asthmatics was significantly suppressed dose-dependently by OKY-046. Secondly, there was a tendency that neutrophil chemotactic factor (NCF) and eosinophil chemotactic factor (ECF) from peripheral blood mononuclear cells stimulated by Candida antigen in intractable asthmatics were suppressed by OKY-046. Thirdly, leukotriene (LTC4) and superoxide (O2-) production from peripheral blood neutrophils in intractable asthmatics was significantly suppressed dose-dependently by OKY-046. That is, OKY-046 has a suppressive effect on type IV allergy caused by lymphocyte activation and on mediator release from neutrophils. These results suggest that TXA2 plays an important role in the development of bronchial asthma and OKY-046 might be a useful drug in the treatment of intractable asthmatics.

Topics: Adult; Aged; Asthma; Depression, Chemical; Dose-Response Relationship, Drug; Female; Humans; Interleukin-2; Interleukin-8; Lymphocyte Activation; Lymphocytes; Male; Methacrylates; Middle Aged; Neutrophils; SRS-A; Superoxides; Thromboxane-A Synthase

Eosinophilia and eosinophil function are regulated by cytokines such as granulocyte/macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and interleukin-5 (IL-5). We have investigated the modulatory role of IL-5 on N-formyl-methionyl-leucyl-phenylalanine (FMLP), neutrophil-activating factor (NAF/IL-8), platelet factor 4 (PF4), and cytokine-induced chemotaxis of eosinophils from normal individuals. These eosinophils show a small chemotactic response toward PF4 but not to NAF/IL-8 and FMLP. Preincubation of eosinophils with low concentrations of IL-5 caused significantly increased responses toward PF4 and induced a significant chemotactic response toward FMLP and NAF/IL-8. In marked contrast, IL-5 (or IL-3) priming of eosinophils from normal donors resulted in a strong inhibition of GM-CSF-induced chemotaxis. A similar decrease in the chemotactic response toward GM-CSF was observed in eosinophils derived from allergic asthmatic individuals. This finding suggests that the latter eosinophils may have had a prior exposure to IL-5 (or IL-3). Washing of the cells after priming did not abrogate the inhibition of the GM-CSF response. Our data indicate that at low concentrations IL-5 is an important modulator of eosinophil chemotaxis, causing selective upregulation or downregulation of chemotactic responses toward different agents.

Topics: Adult; Asthma; Chemotaxis, Leukocyte; Eosinophils; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-3; Interleukin-5; Interleukin-8; Male; Middle Aged; N-Formylmethionine Leucyl-Phenylalanine; Platelet Factor 4

We have previously demonstrated that cultured human bronchial epithelial cells produce cytokines with potent proinflammatory properties on exposure to several stimuli in vitro, and we have hypothesized that these epithelial cell-derived factors may contribute to the pathogenesis of some inflammatory diseases of the bronchial mucosa, particularly asthma, by promoting the infiltration of granulocytes and T cells and their local activation. We provide, in this study, direct evidence of an increased expression of granulocyte-macrophage-colony-stimulating factor, interleukin-6, and interleukin-8 genes and proteins in bronchial epithelium from patients with symptomatic asthma. The up regulation of the production of these cytokines in bronchial epithelial cells of patients with asthma could be abolished in vitro by corticosteroids (hydrocortisone, 10(-7) mol/L), but the up regulation also spontaneously disappeared during a period of 6 days after the removal of the cells from the diseased tissue.

Topics: Adrenal Cortex Hormones; Adult; Asthma; Base Sequence; Bronchi; Cytokines; Epithelium; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunohistochemistry; Interleukin-6; Interleukin-8; Male; Middle Aged; Molecular Probes; Molecular Sequence Data; RNA, Messenger

In order to investigate the mechanism of adenosine-induced bronchoconstriction in asthma, serum neutrophil chemotactic activity (NCA) was measured in normal individuals and patients with asthma before and 5 min after bronchoprovocation testing with adenosine. Challenge testing was terminated when the FEV1 fell by 20% or a concentration of 10 mg/ml was reached. Participants were separated into three groups: six asthmatics hyperresponsive to adenosine (Group 1), seven asthmatics hyperresponsive to histamine but not adenosine (Group 2), and six normal subjects (Group 3). The mean percentage increase in NCA was 84% for Group 1 (p less than 0.001), 29% for Group 2 (p less than 0.05), and only 13% for Group 3. No significant increase in NCA was observed after histamine challenge in seven individuals with asthma derived from Groups 1 and 2. Four patients from Group 1 were rechallenged after treatment with therapeutic doses of oral theophylline. Theophylline therapy was associated with a significant attenuation of the increase in NCA at the concentration of adenosine which caused a 20% decrease in FEV1 before treatment (18% versus 84%, p less than 0.01). The concentration of adenosine which caused a 20% drop in FEV1 was increased at least twofold for each of the four patients. Analysis of NCA by gel filtration chromatography demonstrated an increase in a high molecular weight neutrophil chemotactic factor in the serum of two Group 1 patients after adenosine challenge. Release of a high molecular weight neutrophil chemotactic factor is consistent with a mast cell source for inflammatory mediators in adenosine-induced bronchoconstriction. The therapeutic effects of theophylline, a potent adenosine antagonist, in asthma may therefore occur in part through the inhibition of this process.

Topics: Adenosine; Adult; Asthma; Bronchial Provocation Tests; Bronchoconstriction; Chemotaxis, Leukocyte; Female; Humans; Interleukin-8; Male; Middle Aged; Neutrophils; Theophylline

The mechanism of exercise-induced asthma (EIA) is still controversial, although the role of chemical mediator is strongly suspected. In the present study, 50 asthmatic patients were observed after exercise on bicycle ergometer during dry air breathing, and changes of plasma histamine and neutrophil chemotactic factor (NCF) were measured and effect of anti-allergic drugs was examined. 31 patients developed postexertional bronchoconstriction and their % reduction of FEV1 after exercise correlated significantly with the degree of airway hyperresponsiveness to inhaled methacholine determined by Astograph. Plasma histamine levels were examined in 20 EIA positive cases and 13 EIA negative cases, but no significant changes were observed between pre- and post-histamine levels in either group. On the other hand, NCF was elevated significantly after exercise in both EIA positive and negative cases, but postexertional NCF levels were significantly higher in EIA positive than in EIA negative cases. The relationship between % increase of NCF and the % reduction of FEV1 after exercise was significant (r = 0.472, p less than 0.05). DSCG and Azelastine protected the development of EIA in 14 out on 19 cases and 7 out of 12 cases, respectively. Pretreatment with DSCG significantly reduced the increase of NCF after exercise. These results indicates that one of the chemical mediator, NCF, may play an important role in producing postexertional bronchoconstriction in asthmatic patients.

Topics: Adolescent; Adult; Asthma; Asthma, Exercise-Induced; Chemotactic Factors; Female; Histamine; Histamine H1 Antagonists; Humans; Interleukin-8; Male; Middle Aged; Phthalazines

Heat-stable (HS) and heat-labile (HL) neutrophil chemotactic activities (NCAs) have been demonstrated in serum after allergen challenge of subjects with asthma. In this investigation, we have studied the possible occurrence of similar activities in 20 atopic individuals on natural exposure to allergen, that is, during the birch-pollen season. Since eosinophil accumulation is a hallmark of an ongoing allergic inflammation in the respiratory tract also, the possible production of eosinophil chemotactic activity (ECA) was examined in serum after allergen challenge and at natural exposure to pollen. Both HL-NCA and HL-ECA were produced to a significant extent (p less than 0.001) during the season, with the peak of activities occurring simultaneously with the peak pollen count. HL-ECA was produced after allergen challenge of subjects with asthma in the laboratory, as has been demonstrated for NCA previously. The activity of the HS-NCA was unaltered during season. Gel-filtration studies of the major HL-NCA and HL-ECA indicated a molecular weight for both activities of 100 to 150,000, and the activities produced during season cochromatographed with the HL-NCA and HL-ECA produced after allergen challenge in the laboratory, suggesting that all these activities are due to one and the same molecule. The results suggest that the heat-labile chemotactic activity found in serum of atopic subjects and subjects with asthma after allergen exposure may be involved in the attraction of eosinophils and neutrophils to the site of allergic inflammation.

Topics: Asthma; Chemotactic Factors; Chemotactic Factors, Eosinophil; Chemotaxis, Leukocyte; Chromatography, Gel; Eosinophils; Humans; Interleukin-8; Neutrophils; Pollen

The activation of blood neutrophil leukocytes has been proven in subjects with IgE-mediated and non-IgE-mediated asthma. This event appears to be modulated by the release of humoral factors. We submitted 12 toluene-diisocyanate (TDI) asthmatic workers to TDI provocation. During the late asthmatic reaction there was release of a serum chemoattracting factor for normal neutrophil leukocytes and activation of asthmatic neutrophil leukocytes. This appears to be the first demonstration of neutrophil chemotactic activity liberated during the late TDI reaction in humans. The results are explained by an acute inflammatory process occurring during the late asthmatic reaction induced by TDI.

Topics: Adult; Asthma; Bronchial Provocation Tests; Chemotactic Factors; Chemotaxis, Leukocyte; Cyanates; Female; Humans; Interleukin-8; Male; Neutrophils; Occupational Diseases; Toluene 2,4-Diisocyanate

Topics: Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Chemotactic Factors; Humans; Interleukin-8; Kallikreins; Superoxides

Topics: Air; Asthma; Bronchial Spasm; Chemotactic Factors; Cold Temperature; Humans; Hyperventilation; Interleukin-8; Physical Exertion

Topics: Animals; Asthma; Cells, Cultured; Chemotactic Factors; Humans; Interleukin-8; Lymphocytes; Mites; Molecular Weight; Monocytes; Phytohemagglutinins; Swimming

Topics: Asthma; Asthma, Exercise-Induced; Chemotactic Factors; Humans; Interleukin-8; Lung Volume Measurements; Physical Exertion; Sympathetic Nervous System; Water Loss, Insensible

We have previously shown that there were elevations of neutrophil chemotactic activity (NCA) and increases in the percentages of neutrophil and monocyte complement rosettes after exercise-induced asthma (EIA). These observations suggested that leukocyte activation may occur after EIA, possibly as a result of the release of mast-cell-associated mediators. In the present study, we have attempted to establish whether neutrophils and monocytes are functionally altered after EIA as assessed by changes in their cytotoxic capacity. Cytotoxicity was assessed by a direct visual killing assay using opsonized (complement-coated) schistosomula of Schistosoma mansoni as target organisms. Neutrophils and mononuclear cells obtained from 8 patients after exercise-induced asthma (EIA+ve) had increased cytotoxicity for opsonized schistosomula for as long as 60 min after exercise. These changes were preceded by elevations in the concentrations of serum high molecular weight NCA (which were maximal at 10 min after exercise). In asthmatic patients who did not develop exercise-induced asthma (EIA-ve), no significant increases in neutrophil or mononuclear cell killing of schistosomula, or serum NCA concentrations, were observed. There was a highly significant correlation (p less than 0.001) between the reduction in FEV1 and the increases in neutrophil cytotoxicity. In 5 EIA+ve patients, administration of disodium cromoglycate (cromolyn) prior to the exercise task inhibited both the enhancement in neutrophil and mononuclear cell cytotoxicity, as well as the elevations in circulating NCA and the reductions in FEV1.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adolescent; Adult; Asthma; Asthma, Exercise-Induced; Chemotactic Factors; Cromolyn Sodium; Cytotoxicity, Immunologic; Female; Forced Expiratory Volume; Humans; Interleukin-8; Leukocytes; Phagocytosis; Schistosoma mansoni

To investigate whether exercise- and ultrasonic "fog"-induced asthma are due to the same mechanism, i.e. mediator release induced by osmotic changes, we measured the serum neutrophil chemotactic activity before and after exercise and inhalation of "fog" in 15 asthmatic subjects. To assess changes in airway caliber we measured specific airway conductance (SGaw); to assess changes in neutrophil chemotactic activity we measured the maximum distance reached by neutrophils in a filter when challenged with the subject's serum in a Boyden chamber. In 10 subjects, SGaw decreased by more than 35% and neutrophil chemotactic activity increased significantly (P less than 0.05) both after exercise and "fog", whereas in five subjects no change occurred either after exercise or "fog". We conclude that both exercise- and "fog"-induced asthma are associated with increased serum neutrophil chemotactic activity, and that both stimuli may cause asthma by osmotically triggering mediator release from mast cells.

Topics: Adolescent; Adult; Airway Resistance; Asthma; Asthma, Exercise-Induced; Chemotactic Factors; Female; Humans; Humidity; Interleukin-8; Male; Middle Aged; Neutrophils; Physical Exertion

Adenosine, when it is administered by inhalation to asthmatic subjects, is a potent bronchoconstrictor, although its mechanism of action is not known. Since adenosine has been demonstrated to potentiate IgE-dependent mediator release from mast cells, we have investigated the possible relationship between adenosine-induced bronchoconstriction and release of mast cell mediators in 14 asthmatic subjects. In the first study the effect of the putative mast cell-stabilizing drug cromolyn sodium (SCG) was observed on the dose-related changes in SGaw and FEV1 produced by inhaled adenosine and histamine in seven subjects. Inhaled SCG (20 mg) had no effect on the airway responses to histamine. In contrast SCG significantly protected against adenosine-induced bronchoconstriction in four of the seven subjects as reflected by a decrease in the airway response to the highest concentrations of adenosine, from 65 +/- 8% to 12 +/- 3% (mean +/- SEM) for SGaw and 31 +/- 7% to 8 +/- 3% for FEV1. Those three subjects whose adenosine response was unaffected by SCG had received regular SCG until 12 hr before the studies. In a separate study on eight subjects, a single inhalation of adenosine, causing a maximum 61 +/- 4% fall in SGaw at 10 min, had no significant effect on circulating levels of histamine, neutrophil chemotactic factor, or cyclic AMP. Together these two studies suggest that bronchoconstriction produced by adenosine is not a consequence of enhanced mast cell-mediator release and that the inhibitory effects of SCG occur by a mechanism other than through mast cell stabilization.

Topics: Adenosine; Adult; Asthma; Bronchial Provocation Tests; Bronchial Spasm; Chemotactic Factors; Cromolyn Sodium; Cyclic AMP; Female; Forced Expiratory Volume; Histamine; Humans; Interleukin-8; Male; Mast Cells; Middle Aged

Topics: Adolescent; Asthma; Asthma, Exercise-Induced; Chemotactic Factors; Child; Female; Forced Expiratory Volume; Humans; Interleukin-8; Male; Pulse; Swimming

Circulating concentrations of the mast cell-associated mediators, histamine and neutrophil chemotactic factor (NCF) of high molecular weight, were measured in atopic and nonatopic asthmatics after treadmill exercise. Elevations in the concentrations of both mediators accompanied the development of exercise-induced asthma (EIA). Normal individuals did not release mediators or develop bronchoconstriction after an identical exercise. The elaboration of mediators was not due to the onset of airflow obstruction, the postexercise basophilia, or the exercise task per se. A treadmill exercise undertaken while inhaling fully conditioned air inhibited EIA and NCF release; in contrast the same exercise undertaken while breathing cold, dry air elicited EIA and the production of mediators. This suggests that the stimulus for EIA and mediator release may be identical. Late-phase asthmatic reactions occur 3 to 9 hr after exercise in some asthmatics and are accompanied by the appearance of circulating NCF, as previously reported in allergen-induced late responses. In addition to the contribution of mediators to the spasmogenic reaction in EIA, mediators may contribute to bronchial inflammation by activating circulating leukocytes. There was a kinetic increase in the expression of neutrophil C3b receptors in EIA (+) asthmatics for up to 60 min after treadmill exercise. The enhancement of C3b receptors, as evidence of neutrophil activation, was preceded by release of NCF and reductions in peak expiratory flow rates. The prior administration of cromolyn inhibited EIA, NCF release, and enhancement of C3b receptors. These changes were not observed in EIA (-) asthmatics after an identical exercise task. These findings support the view that mediators are released in EIA and may play an important role in its pathogenesis.

Topics: Asthma; Asthma, Exercise-Induced; Basophils; Chemotactic Factors; Cromolyn Sodium; Female; Histamine; Histamine Release; Humans; Hypersensitivity, Immediate; Interleukin-8; Male; Time Factors

Immediate and late asthmatic reactions may occur after inhalation of specific antigens such as house dust mites or pollens. Recently, similar dual reactions have been found to occur after asthma induced by exercise. The immediate reaction to an antigen is believed to be due to the release of preformed and newly formed mediators derived primarily from mast cells via IgE antibodies. The late reaction also appears to be the result of an IgE reaction but involves inflammation in which cells play a role. The late reaction to antigen is often more severe than the immediate reaction and may be followed by a prolonged period of "nonspecific" bronchial hyperreactivity. By contrast, the late reaction to exercise is usually less severe than the immediate reaction. Both immediate and late exercise reactions are also associated with peaks of neutrophil chemotactic factor activity. Cromolyn sodium administered before challenge modifies both the immediate and the late response to antigen and exercise. Glucocorticoids block the late but not the early reaction to antigens. Their effect on the late exercise reaction is not yet known. Albuterol prevents the initial reaction to both exercise and antigen and may modify the late reaction to exercise as well.

Topics: Albuterol; Antigens; Asthma; Asthma, Exercise-Induced; Bronchial Provocation Tests; Chemotactic Factors; Cromolyn Sodium; Forced Expiratory Volume; Histamine Release; Humans; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Interleukin-8; Mast Cells; Methacholine Chloride; Methacholine Compounds; Respiratory Hypersensitivity; Skin Tests; Theophylline

Extreme sensitivity of airways to multiple stimuli characterizes asthma. Airway hyperresponsiveness can be produced experimentally in otherwise healthy subjects or animals by inflammatory damage (e.g., induced by respiratory viruses or by inhaled oxidants). Evidence is presented that cell-to-cell interactions play an important role in experimental hyperreactivity and that similar inflammatory cascades may play a similar role in clinical asthma. Although the importance of epithelial cells and neutrophils has been identified in the present studies, other inflammatory mechanisms (e.g., sensory nerve release of substance P, epithelial mast cells, eosinophils) may also play key roles. In exercise-induced bronchospasm, the stimulus (e.g., cooling or drying) must affect a cell (e.g., one near the epithelial surface) by decreasing temperature or by increasing osmolality. This signal may cause mediator release and a subsequent cascade, leading to contraction of smooth muscle. Environmental irritants (e.g., ozone) inhaled during exercise may potentiate these effects by producing further inflammation.

Topics: Animals; Arachidonic Acids; Asthma; Cell Communication; Chemotactic Factors; Dinoprostone; Dogs; Epithelial Cells; Epithelium; Humans; Hydroxyeicosatetraenoic Acids; Inflammation; Interleukin-8; Leukotriene B4; Neutrophils; Ozone; Prostaglandins E; Sputum

In order to determine if mast cell mediators are released during aspirin challenge in aspirin-sensitive asthmatics, we measured neutrophil chemotactic activity, which has been shown to be an indicator of mast cell degranulation. Four aspirin-sensitive asthmatic subjects were given doses of aspirin (60 to 325 mg) previously determined to cause a 20 to 30% fall in forced expiratory volume in one second (FEV1); pulmonary function was followed by serial spirograms and body plethysmography. Serum was obtained before and at 30, 60, 90, 120, 180, and 240 min after challenge, corresponding to the times of pulmonary function measurements. Neutrophil chemotactic activity was measured using a modified Boyden chamber assay. Maximal bronchoconstriction occurred 60 to 120 min after aspirin ingestion. An increase in neutrophil chemotactic activity of 300 to 600% over baseline was detected in all subjects. In 3 subjects, neutrophil chemotactic activity release paralleled bronchoconstriction, and in 1 subject, it followed onset of bronchoconstriction. Physicochemical analysis showed that the neutrophil chemotactic activity eluted in the void volume of a Sephadex G-200 column (greater than or equal to 250,000 daltons) and from Sephadex QAE anion exchange chromatography in a region corresponding to 0.2 to 0.3 M NaCl. Its isoelectric point was in the pH range 6.5 to 7.5. These characteristics are compatible with neutrophil chemotactic factor of mast cell origin. Pretreatment with sodium cromolyn (40 mg) completely eliminated neutrophil chemotactic factor release, but only partially suppressed the fall in FEV1 in 2 subjects and had no effect on FEV1 fall in a third.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Aspirin; Asthma; Chemotactic Factors; Chemotaxis, Leukocyte; Chromatography, Gel; Chromatography, Ion Exchange; Cromolyn Sodium; Cytoplasmic Granules; Female; Forced Expiratory Volume; Humans; Interleukin-8; Isoelectric Focusing; Male; Mast Cells; Middle Aged; Neutrophils

Neutrophil chemotactic activity (NCA) following oral challenge with aspirin (ASA) was determined in ASA-intolerant asthmatic subjects, and in ASA-tolerant asthmatic and normal subjects. There was a statistically significant fall in FEV1 and a rise in NCA (P less than 0.01) following challenge in the ASA-sensitive subjects compared with that of the ASA-tolerant subjects and normal controls. No significant difference was observed between the latter two groups. The chemotactic factor identified in this study had a molecular weight greater than 150 000 which is consistent with NCF-A (neutrophil chemotactic factor of anaphylaxis). The ASA-induced fall in FEV1 and rise in NCA was further studied in three of the ASA-intolerant asthmatic subjects, with and without pretreatment with inhaled sodium cromoglycate. In these subjects, the drug inhibited both the oral ASA-induced bronchoconstriction and the increase in neutrophil chemotactic activity. These results suggest that ASA-induced asthma involves mediator release from mast cells, as shown by the increase in NCA following ASA challenge and the protective effect of sodium cromoglycate which is considered to inhibit mast-cell degranulation.

Topics: Adult; Aspirin; Asthma; Chemotactic Factors; Cromolyn Sodium; Drug Hypersensitivity; Female; Forced Expiratory Volume; Humans; Interleukin-8; Male; Mast Cells

Two adults and 13 children with exercise-induced asthma had both immediate and late reductions in forced expiratory volume in one second (FEV1) after treadmill exercise. The late reactions developed 4 to 10 hours after exercise and in each instance were associated with wheezing or chest tightness (or both). Increases in neutrophil chemotactic activity, measured in the 2 adults and in 11 of the children, accompanied the reductions in FEV1 in all these subjects. In contrast, four other adults with only an immediate fall in FEV1 after exercise had only an initial elevation in neutrophil chemotactic activity, with no subsequent increase for the remaining 24-hour period. The agent responsible for the neutrophil chemotactic activity released during exercise-induced late reactions appeared to be identical to that released during immediate reactions. These observations suggest that some patients with exercise-induced asthma have late reactions that, as in the case of antigen-induced bronchoconstriction, are accompanied by the release of neutrophil chemotactic activity.

Topics: Adolescent; Adult; Asthma; Asthma, Exercise-Induced; Chemotactic Factors; Chemotaxis, Leukocyte; Child; Chromatography, Gel; Female; Forced Expiratory Volume; Humans; Interleukin-8; Male; Neutrophils

Topics: Animals; Asthma; Bronchi; Bronchial Provocation Tests; Chemotactic Factors; Forced Expiratory Volume; Histamine; Humans; Interleukin-8; Lung; Rabbits; Time Factors

Elevated levels of the mast-cell-associated serum neutrophil chemotactic factor (NCF) and an increase in blood basophil counts were observed in 6 atopic asthmatics during exercise-induced asthma (EIA). These changes were not found when the same degree of airways obstruction was elicited in the same subjects by isocapnic hyperventilation (ISH) with cold air. The NCF was unlikely to be related to the basophilia alone, because asthmatics without EIA who underwent the same exercise task, produced a similar basophilia but significantly less NCF. These findings suggest that mast-cell-associated (as opposed to basophil-associated) mediators of hypersensitivity are detectable in the bloodstream during the bronchoconstriction induced by exercise, but not by ISH.

Topics: Adolescent; Adult; Asthma; Asthma, Exercise-Induced; Basophils; Chemotactic Factors; Cold Temperature; Female; Humans; Hyperventilation; Interleukin-8; Leukocyte Count; Male; Mast Cells; Physical Exertion

Neutrophil chemotactic factor (NCF) is a slightly acidic macromolecule which is released into the circulation of asthmatic individuals following an exercise task. Its appearance was basophil-independent, accompanied the development of airflow obstruction and its release could be inhibited by administration of disodium cromoglycate. There was no elaboration of NCF after isocapnic hyperventilation with cold air, although the same asthmatics produced NCF following treadmill exercise. Finally, NCF was also released during exercise-induced late asthmatic reactions, as in antigen-induced late responses.

Topics: Asthma; Asthma, Exercise-Induced; Basophils; Chemotactic Factors; Humans; Hyperventilation; Interleukin-8; Respiratory Function Tests

Topics: Asthma; Asthma, Exercise-Induced; Bronchial Provocation Tests; Chemotactic Factors; Cromolyn Sodium; Forced Expiratory Volume; Histamine; Humans; Interleukin-8

Concentrations of plasma histamine and serum neutrophil chemotactic factor (NCF) were measured in seven atopic asthmatics who developed exercise-induced asthma (EIA) after a treadmill task. The results were compared with those obtained after inhalation of specific antigen or methacholine. Plasma histamine concentrations were measured with a novel double-isotope radiometric assay, and NCF was identified by its elution in the void volume fractions of Sephadex G-200 and as a single peak of activity at approximately 0.20 molar NaCl after anion exchange chromatography on diethylaminoethyl-Sephacel (pH 7.8). After exercise or antigen challenge, the time courses of appearance of both mediators were virtually identical and accompanied the increase in airways obstruction. There was a statistically significant correlation between the concentrations of histamine or NCF and the magnitude of airflow obstruction after exercise and antigen challenge. This suggested that there may be a direct association between mediator release and EIA or antigen-induced bronchoconstriction. In contrast, there were no significant elevations in circulating histamine and NCF after inhalation of methacholine, at concentrations giving a fall in FEV1 comparable to that induced by exercise or antigen. The prior administration of cromolyn to three asthmatics inhibited both their EIA and the release of histamine and NCF. When four asthmatics were exercised for periods of 1, 3, and 6 min, the release of NCF and fall in peak expiratory flow rate were directly related to the duration of the exercise. The rise of NCF activity in subjects with EIA was fivefold greater than that observed in asthmatics who did not experience airways obstruction when subjected to the same exercise task. These results provide further evidence that mediators of hypersensitivity are released during EIA.

Topics: Adolescent; Adult; Asthma; Chemotactic Factors; Female; Histamine Release; Humans; Immunoglobulin E; Interleukin-8; Male; Physical Exertion; Skin Tests

Topics: Adolescent; Adult; Allergens; Antigen-Antibody Complex; Asthma; Bronchial Provocation Tests; Chemotactic Factors; Female; Hot Temperature; Humans; Interleukin-8; Lactoferrin; Macrophages; Male; Monocytes; Muramidase; Neutrophils; Peak Expiratory Flow Rate

Significant increase in the maximum post-exercise values of plasma histamine (PH), whole blood histamine (WBH) and neutrophil chemotactic factor (NCF) occurred in arterial blood within the first hour after exercise in asthmatic patients. However, similar changes in PH and WBH also occurred in the control group. Significant increases in circulating basophil counts following exercise were found in both groups, which closely mirrored the changes in PH and NCF, and there was a highly significant correlation between rises in WBH and basophil counts (P less than 0.001). When plasma histamine as assayed in venous blood using a more sensitive and specific double isotope radio enzymatic assay no significant alteration in plasma histamine levels was detected in either the asthmatic or the control group. We conclude that there is no evidence from these studies to support the suggestion that mast cell mediator release is involved in the pathogenesis of exercise-induced asthma, and that any observed changes in levels of PH and NCF after exercise may be related to changes in levels of circulating basophils.

Topics: Adult; Asthma; Asthma, Exercise-Induced; Basophils; Chemotactic Factors; Forced Expiratory Volume; Histamine; Humans; Interleukin-8; Leukocyte Count; Male; Peak Expiratory Flow Rate; Respiratory Function Tests

Topics: Asthma; Basophils; Bronchi; Chemotactic Factors; Chemotaxis, Leukocyte; Eosinophils; Humans; Interleukin-8; Leukotriene B4; Mast Cells; Neutrophils

Topics: Antigens; Asthma; Bronchial Spasm; Chemotactic Factors; Chemotactic Factors, Eosinophil; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Interleukin-8; Lymphocytes; Mast Cells; Neutrophils