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1.
Clin Exp Allergy ; 47(4): 488-498, 2017 Apr.
Article in English | MEDLINE | ID: mdl-28000949

ABSTRACT

BACKGROUND: Eosinophils contribute to the pathogenesis of multiple diseases, including asthma. Treatment with antibodies targeting IL-5 or IL-5 receptor α reduces the frequency of asthma exacerbations. Eosinophil receptors for IL-5 share a common ß-chain with IL-3 and GM-CSF receptors. We recently reported that IL-3 is more potent than IL-5 or GM-CSF in maintaining the ERK/p90S6K/RPS6 ribosome-directed signaling pathway, leading to increased protein translation. OBJECTIVE: We aimed to determine disease-relevant consequences of prolonged eosinophil stimulation with IL-3. RESULTS: Human blood eosinophils were used to establish the impact of activation with IL-3 on IgG-driven eosinophil degranulation. When compared to IL-5, continuing exposure to IL-3 further induced degranulation of eosinophils on aggregated IgG via increased production and activation of both CD32 (low affinity IgG receptor) and αMß2 integrin. In addition, unlike IL-5 or GM-CSF, IL-3 induced expression of CD32B/C (FCGRIIB/C) subtype proteins, without changing CD32A (FCGRIIA) protein and CD32B/C mRNA expression levels. Importantly, these in vitro IL-3-induced modifications were recapitulated in vivo on airway eosinophils. CONCLUSIONS AND CLINICAL RELEVANCE: We observed for the first time upregulation of CD32B/C on eosinophils, and identified IL-3 as a potent inducer of CD32- and αMß2-mediated eosinophil degranulation.


Subject(s)
Cell Degranulation/immunology , Eosinophils/immunology , Eosinophils/metabolism , Interleukin-3/metabolism , Macrophage-1 Antigen/metabolism , Receptors, IgG/metabolism , Antibodies, Monoclonal/pharmacology , Biomarkers , Cell Degranulation/drug effects , Cells, Cultured , Eosinophils/drug effects , Humans , Interleukin-3/pharmacology , Receptors, IgG/antagonists & inhibitors
2.
Clin Exp Allergy ; 44(12): 1484-93, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25109477

ABSTRACT

BACKGROUND: Asthma exacerbations contribute to significant morbidity, mortality and healthcare utilization. Furthermore, viral infections are associated with asthma exacerbations by mechanisms that are not fully understood. OBJECTIVE: The aim of this analysis was to determine whether cytokine patterns in patients with colds could identify risks for subsequent asthma exacerbations. METHODS: We analysed cytokine levels in nasal lavage fluid (NLF) in 59 subjects (46 with asthma) with acute upper respiratory symptoms and after symptomatic resolution. Analyte choice was based on potential relevance to asthma exacerbations: antiviral (IFN-α, IFN-ß, IFN-γ, IFN-λ1, IP-10, TRAIL), cell recruiting (G-CSF, IL-1ß, IL-8, MCP-1, MCP-3, TNF-α), polarizing (CXCL13, IL-10, IL-13, IL-17, TSLP), and injury remodelling (fibronectin, IL-33, MMP-9, VEGF). RESULTS: The overall cytokine response induced during viral infections was not different between asthmatic and non-asthmatic individuals for a wide array of cytokines. However, mean levels of VEGF, TNF-α and IL-1ß were 1.7-, 5.1- and 4.7-fold higher in samples from asthma subjects who exacerbated in the first 3 weeks of the cold compared with those who did not exacerbate (P = 0.006, 0.01, 0.048, respectively). Using receiver operating characteristic curve-defined thresholds, high VEGF and TNF-α levels predicted a shorter time-to-exacerbation after NLF sampling (25% exacerbation rate: 3 vs. 45 days, and 3 vs. 26 days; P = 0.03, 0.04, respectively). CONCLUSION AND CLINICAL RELEVANCE: Although they produce similar cytokine responses to viral infection as non-asthmatics, asthmatics with higher levels of VEGF and TNF-α in NLF obtained during acute cold phases predicted subsequent asthma exacerbations in this cohort of patients with mild-to-moderate disease. In the future, stratifying the risk of an asthma exacerbation by cytokine profile may aid the targeting of personalized treatment and intervention strategies.


Subject(s)
Asthma/diagnosis , Asthma/etiology , Common Cold/complications , Common Cold/metabolism , Nasal Lavage Fluid , Tumor Necrosis Factor-alpha/metabolism , Vascular Endothelial Growth Factor A/metabolism , Cytokines/metabolism , Disease Progression , Female , Humans , Male , Nasal Lavage Fluid/immunology , ROC Curve
3.
Clin Exp Allergy ; 44(6): 813-21, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24450586

ABSTRACT

BACKGROUND: The majority of asthma exacerbations are related to viral respiratory infections. Some, but not all, previous studies have reported that low interferon responses in patients with asthma increase the risk for virus-induced exacerbations. OBJECTIVE: We sought to determine the relationship between lower airway inflammatory biomarkers, specifically interferon gene expression, and the severity or presence of an exacerbation in asthmatics experiencing a naturally occurring viral infection. METHODS: Sputum samples were analysed from subjects in an asthma exacerbation study who experienced a confirmed viral infection. Subjects were monitored for daily symptoms, medication use and peak expiratory flow rate until baseline. Sputum samples were assessed for cell counts and gene expression. RESULTS: Interferon gamma expression was significantly greater in patients with asthma exacerbations compared to non-exacerbating patients (P = 0.002). IFN-α1, IFN-ß1 and IFN-γ mRNA levels correlated with the peak Asthma Index (r = 0.58, P < 0.001; r = 0.57, P = 0.001; and r = 0.51, P = 0.004, respectively). Additionally, IL-13, IL-10 and eosinophil major basic protein mRNA levels were greater in patients with asthma exacerbations compared to non-exacerbating patients (P = 0.03, P = 0.06 and P = 0.02, respectively), and IL-13 mRNA correlated with the peak Asthma Index (P = 0.006). CONCLUSIONS: Our findings indicate that asthma exacerbations are associated with increased rather than decreased expression of interferons early in the course of infection. These findings raise the possibility that excessive virus-induced interferon production during acute infections can contribute to airway inflammation and exacerbations of asthma.


Subject(s)
Asthma/genetics , Asthma/physiopathology , Gene Expression , Interferons/genetics , Sputum/cytology , Adult , Asthma/complications , Biomarkers , Cytokines/genetics , Disease Progression , Female , Humans , Male , Middle Aged , RNA, Messenger/genetics , Respiratory Function Tests , Severity of Illness Index , Virus Diseases/complications , Virus Diseases/diagnosis , Young Adult
4.
Clin Exp Allergy ; 44(1): 38-46, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24131304

ABSTRACT

BACKGROUND: Interleukin 13 (IL13) is a T-helper type 2 (Th2) cytokine associated with inflammation and pathology in allergic diseases such as bronchial asthma. We have shown that treatment with lebrikizumab, an anti-IL13 monoclonal antibody, significantly improves prebronchodilator forced expiratory volume in 1 s (FEV(1)) in a subset of subjects with uncontrolled asthma. OBJECTIVE: To evaluate efficacy and safety of lebrikizumab in subjects with mild asthma who underwent bronchial allergen challenge. METHODS: Twenty-nine subjects were randomized 1 : 1-5 mg/kg lebrikizumab (n = 13) or placebo (n = 16) administered subcutaneously every 4 weeks over 12 weeks, a total of four doses. Primary efficacy outcome was late asthmatic response (LAR) at Week 13, defined as area under the curve of FEV1 measured 2-8 h following inhaled allergen challenge. Serum biomarkers were measured to verify IL13 pathway inhibition and identify patients with an increased response to lebrikizumab. RESULTS: At Week 13, the LAR in lebrikizumab subjects was reduced by 48% compared with placebo subjects, although this was not statistically significant (95% confidence interval, -19%, 90%). Exploratory analysis indicated that lebrikizumab-treated subjects with elevated baseline levels of peripheral blood eosinophils, serum IgE, or periostin exhibited a greater reduction in LAR compared with subjects with lower baseline levels of these biomarkers. Lebrikizumab exerted systemic effects on markers of Th2 inflammation, reducing serum immunoglobulin E (IgE), chemokine ligands 13 and 17 by approximately 25% (P < 0.01). Lebrikizumab was well tolerated. CONCLUSION AND CLINICAL RELEVANCE: Lebrikizumab reduced the LAR in subjects with mild asthma. Clinical trial number NCT00781443.


Subject(s)
Allergens/immunology , Anti-Asthmatic Agents/therapeutic use , Antibodies, Monoclonal/therapeutic use , Asthma/drug therapy , Asthma/immunology , Adult , Anti-Asthmatic Agents/adverse effects , Anti-Asthmatic Agents/pharmacology , Antibodies, Monoclonal/adverse effects , Antibodies, Monoclonal/pharmacology , Asthma/blood , Biomarkers/blood , Bronchial Provocation Tests , Female , Forced Expiratory Volume/drug effects , Humans , Interleukin-13 , Lung/immunology , Lung/physiopathology , Male , Middle Aged , Th2 Cells/immunology , Th2 Cells/metabolism , Treatment Outcome , Young Adult
5.
Clin Exp Allergy ; 43(12): 1342-50, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24261944

ABSTRACT

BACKGROUND: Eosinophilia is a marker of corticosteroid responsiveness and risk of exacerbation in asthma; although it has been linked to submucosal matrix deposition, its relationship with other features of airway remodelling is less clear. OBJECTIVE: The aim of this study was to investigate the relationship between airway eosinophilia and airway remodelling. METHODS: Bronchial biopsies from subjects (n = 20 in each group) with mild steroid-naïve asthma, with either low (0-0.45 mm(-2)) ) or high submucosal eosinophil (23.43-46.28 mm(-2) ) counts and healthy controls were assessed for in vivo epithelial damage (using epidermal growth factor receptor staining), mucin expression, airway smooth muscle (ASM) hypertrophy and inflammatory cells within ASM. RESULTS: The proportion of in vivo damaged epithelium was significantly greater (P = 0.02) in the high-eosinophil (27.37%) than the low-eosinophil (4.14%) group. Mucin expression and goblet cell numbers were similar in the two eosinophil groups; however, MUC-2 expression was increased (P = 0.002) in the high-eosinophil group compared with controls. The proportion of submucosa occupied by ASM was higher in both asthma groups (P = 0.021 and P = 0.046) compared with controls. In the ASM, eosinophil and T-lymphocyte numbers were higher (P < 0.05) in the high-eosinophil group than both the low-eosinophil group and the controls, whereas the numbers of mast cells were increased in the high-eosinophil group (P = 0.01) compared with controls. CONCLUSION: Submucosal eosinophilia is a marker (and possibly a cause) of epithelial damage and is related to infiltration of ASM with eosinophils and T lymphocytes, but is unrelated to mucus metaplasia or smooth muscle hypertrophy.


Subject(s)
Airway Remodeling , Asthma/immunology , Asthma/pathology , Eosinophilia/pathology , Adult , Asthma/metabolism , Case-Control Studies , Female , Goblet Cells/pathology , Humans , Hyperplasia , Male , Middle Aged , Mucins/metabolism , Muscle, Smooth/metabolism , Muscle, Smooth/pathology , Respiratory Mucosa/immunology , Respiratory Mucosa/metabolism , Respiratory Mucosa/pathology , Young Adult
6.
Clin Exp Allergy ; 43(3): 292-303, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23414537

ABSTRACT

BACKGROUND: IL-5 activates α(M) ß(2) integrin on blood eosinophils in vitro. Eosinophils in bronchoalveolar lavage (BAL) following segmental antigen challenge have activated ß(2) -integrins. OBJECTIVE: To identify roles for IL-5 in regulating human eosinophil integrins in vivo. METHODS: Blood and BAL eosinophils were analysed by flow cytometry in ten subjects with allergic asthma who underwent a segmental antigen challenge protocol before and after anti-IL-5 administration. RESULTS: Blood eosinophil reactivity with monoclonal antibody (mAb) KIM-127, which recognizes partially activated ß(2) -integrins, was decreased after anti-IL-5. Before anti-IL-5, surface densities of blood eosinophil ß(2) , α(M) and α(L) integrin subunits increased modestly post challenge. After anti-IL-5, such increases did not occur. Before or after anti-IL-5, surface densities of ß(2) , α(M) , α(L) and α(D) and reactivity with KIM-127 and mAb CBRM1/5, which recognizes high-activity α(M) ß(2) , were similarly high on BAL eosinophils 48 h post-challenge. Density and activation state of ß(1) -integrins on blood and BAL eosinophils were not impacted by anti-IL-5, even though anti-IL-5 ablated a modest post-challenge increase on blood or BAL eosinophils of P-selectin glycoprotein ligand-1 (PSGL-1), a receptor for P-selectin that causes activation of ß(1) -integrins. Forward scatter of blood eosinophils post-challenge was less heterogeneous and on the average decreased after anti-IL-5; however, anti-IL-5 had no effect on the decreased forward scatter of eosinophils in post-challenge BAL compared with eosinophils in blood. Blood eosinophil KIM-127 reactivity at the time of challenge correlated with the percentage of eosinophils in BAL post-challenge. CONCLUSION AND CLINICAL RELEVANCE: IL-5 supports a heterogeneous population of circulating eosinophils with partially activated ß(2) -integrins and is responsible for up-regulation of ß(2) -integrins and PSGL-1 on circulating eosinophils following segmental antigen challenge but has minimal effects on properties of eosinophils in BAL. Dampening of ß(2) -integrin function of eosinophils in transit to inflamed airway may contribute to the decrease in lung inflammation caused by anti-IL-5.


Subject(s)
Antibodies, Monoclonal/immunology , Antigens/immunology , CD18 Antigens/metabolism , Eosinophils/immunology , Eosinophils/metabolism , Interleukin-5/immunology , Adult , Antibodies, Blocking/immunology , Antibodies, Blocking/pharmacology , Antibodies, Monoclonal/pharmacology , Asthma/immunology , Asthma/metabolism , Bronchoalveolar Lavage Fluid/cytology , Bronchoalveolar Lavage Fluid/immunology , CD18 Antigens/immunology , Eosinophils/drug effects , Epitopes/immunology , Female , Humans , Interleukin-5/antagonists & inhibitors , Leukocyte Count , Male , Membrane Glycoproteins/metabolism , Young Adult
7.
Clin Exp Allergy ; 43(2): 187-97, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23331560

ABSTRACT

BACKGROUND: Allergic airway inflammation contributes to the airway remodelling that has been linked to increased obstruction and morbidity in asthma. However, the mechanisms by which allergens contribute to airway remodelling in humans are not fully established. CCL18, chitotriosidase (CHIT1) and YKL-40 are readily detectable in the lungs and contribute to remodelling in other fibrotic diseases, but their involvement in allergic asthma is unclear. OBJECTIVE: We hypothesized that CCL18, YKL-40 and CHIT1 bioactivity are enhanced in allergic asthma subjects after segmental allergen challenge and are related to increased pro-fibrotic and Th2-associated mediators in the lungs. METHODS: Levels of CCL18 and YKL-40 protein and chitotriosidase (CHIT1) bioactivity in bronchoalveolar lavage (BAL) fluid, as well as CCL18, YKL-40 and CHIT1 mRNA levels in BAL cells were evaluated in patients with asthma at baseline and 48 h after segmental allergen challenge. We also examined the correlation between CCL18 and YKL-40 levels and CHIT1 activity with the levels of other pro-fibrotic factors and chemokines previously shown to be up-regulated after allergen challenge. RESULTS: Chitotriosidase activity and YKL-40 and CCL18 levels were elevated after segmental allergen challenge and these levels correlated with those of other pro-fibrotic factors, T cell chemokines, and inflammatory cells after allergen challenge. CCL18 and YKL-40 mRNA levels also increased in BAL cells after allergen challenge. CONCLUSIONS AND CLINICAL RELEVANCE: Our results suggest that CCL18 and YKL-40 levels and CHIT1 activity are enhanced in allergic airway inflammation and thus may contribute to airway remodelling in asthma.


Subject(s)
Allergens/immunology , Asthma/immunology , Asthma/metabolism , Chemokines, CC/metabolism , Chitinases/metabolism , Adipokines/metabolism , Adult , Airway Remodeling , Allergens/administration & dosage , Asthma/genetics , Bronchial Provocation Tests , Bronchoalveolar Lavage Fluid/cytology , Bronchoalveolar Lavage Fluid/immunology , Chitinase-3-Like Protein 1 , Cytokines/metabolism , Enzyme Activation , Female , Humans , Inflammation Mediators/metabolism , Lectins/metabolism , Male , Time Factors , Young Adult
8.
Clin Exp Allergy ; 42(12): 1756-64, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23181791

ABSTRACT

BACKGROUND: Differentiation and activation of CD4(+) T cells is controlled by various cytokines produced by innate immune cells. We have shown that eosinophils (EOS) have the potential to influence Th1 and Th2 cytokine generation by CD4(+) cells, but their influence on IL-17A (IL-17) has not been established. OBJECTIVE: The purpose of this study is to determine the effect of EOS on IL-17 production by lymphocytes. METHODS: Pre-activated CD4(+) T cells were cultured in the presence of either autologous EOS or EOS culture supernatants. Expression of IL-17 was determined by real-time quantitative PCR (qPCR) after 5 h and protein level was measured after 48 h. To determine the effect of allergen-induced airway EOS on IL-17, subjects with mild allergic asthma underwent bronchoscopic segmental bronchoprovocation with allergen (SBP-Ag) after a treatment with an anti-IL-5 neutralizing antibody (mepolizumab) to reduce airway eosinophilia. IL-17 mRNA was measured in bronchoalveolar lavage (BAL) cells by qPCR. RESULTS: In vitro, EOS significantly increased IL-17 production by CD4(+) T cells. Addition of exogenous IL-1ß increased expression of IL-17 mRNA by CD4(+) T cells. EOS expressed and released IL-1ß. Furthermore, levels of IL-1ß in EOS supernatants highly correlated with their ability to increase IL-17 expression by CD4(+) T cells, and neutralizing antibody to IL-1ß reduced expression of IL-17 mRNA. In vivo, reduction of EOS in the airway using mepolizumab was associated with diminished IL-17 expression after SBP-Ag. CONCLUSIONS AND CLINICAL RELEVANCE: Our data demonstrate that EOS can promote IL-17 production through the release of IL-1ß. Enhanced IL-17 cytokine production is another mechanism by which EOS may participate in pathogenesis of allergic airway inflammation in asthma.


Subject(s)
Asthma/immunology , CD4-Positive T-Lymphocytes/immunology , Eosinophils/metabolism , Gene Expression Regulation , Interleukin-17/metabolism , Interleukin-1beta/metabolism , Lymphocyte Activation/immunology , Antibodies, Monoclonal, Humanized/therapeutic use , Asthma/therapy , CD4-Positive T-Lymphocytes/metabolism , Cells, Cultured , Cytokines/genetics , Cytokines/metabolism , Eosinophils/immunology , Gene Expression Regulation/immunology , Humans , Hypersensitivity , Interleukin-17/genetics , Interleukin-1beta/genetics , Treatment Outcome , Up-Regulation
9.
Mucosal Immunol ; 3(1): 69-80, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19710636

ABSTRACT

Rhinovirus (RV) infections trigger asthma exacerbations. Genome-wide expression analysis of RV1A-infected primary bronchial epithelial cells from normal and asthmatic donors was performed to determine whether asthma is associated with a unique pattern of RV-induced gene expression. Virus replication rates were similar in cells from normal and asthmatic donors. Overall, RV downregulated 975 and upregulated 69 genes. Comparisons of transcriptional profiles generated from microarrays and confirmed by quantitative reverse transcription PCR and cluster analysis showed some up- and downregulated genes in asthma cells involved in immune responses (IL1B, IL1F9, IL24, and IFI44) and airway remodeling (LOXL2, MMP10, FN1). Notably, most of the asthma-related differences in RV-infected cells were also present in the cells before infection. These findings suggest that differences in RV-induced gene expression profiles of cells from normal and mild asthmatic subjects could affect the acute inflammatory response to RV, and subsequent airway repair and remodeling.


Subject(s)
Asthma/immunology , Picornaviridae Infections/immunology , Respiratory Mucosa/metabolism , Rhinovirus/physiology , Adult , Airway Remodeling/genetics , Amino Acid Oxidoreductases/biosynthesis , Amino Acid Oxidoreductases/genetics , Asthma/complications , Asthma/genetics , Asthma/pathology , Cells, Cultured , Cytokines/biosynthesis , Cytokines/genetics , Female , Gene Expression Profiling , Gene Expression Regulation , Genome-Wide Association Study , Humans , Male , Matrix Metalloproteinase 10/biosynthesis , Matrix Metalloproteinase 10/genetics , Microarray Analysis , Picornaviridae Infections/complications , Picornaviridae Infections/genetics , Picornaviridae Infections/pathology , Respiratory Mucosa/immunology , Respiratory Mucosa/pathology , Respiratory Mucosa/virology , Virus Replication
10.
Eur Respir J ; 32(6): 1548-54, 2008 Dec.
Article in English | MEDLINE | ID: mdl-18768579

ABSTRACT

The asthmatic response to the common cold is highly variable, and early characteristics that predict worsening of asthma control following a cold have not been identified. In this prospective multicentric cohort study of 413 adult subjects with asthma, the mini-Asthma Control Questionnaire (mini-ACQ) was used to quantify changes in asthma control and the Wisconsin Upper Respiratory Symptom Survey-21 (WURSS-21) to measure cold severity. Univariate and multivariable models were used to examine demographic, physiological, serological and cold-related characteristics for their relationship to changes in asthma control following a cold. Clinically significant worsening of asthma control was observed following a cold (mean+/-SD increase in mini-ACQ score of 0.69+/-0.93). Univariate analysis demonstrated that season, centre location, cold duration and cold severity measurements were all associated with a change in asthma control. Multivariable analysis of the covariates available within the first 2 days of cold onset revealed that the day 2 and cumulative sum of day 1 and 2 WURSS-21 scores were significant predictors of the subsequent changes in asthma control. In asthmatic subjects, cold severity within the first 2 days can be used to predict subsequent changes in asthma control. This information may help clinicians prevent deterioration in asthma control following a cold.


Subject(s)
Asthma/diagnosis , Asthma/physiopathology , Common Cold/complications , Adrenal Cortex Hormones/therapeutic use , Adult , Asthma/etiology , Cohort Studies , Female , Humans , Male , Middle Aged , Multivariate Analysis , Prospective Studies , Quality of Life , Risk , Surveys and Questionnaires , Treatment Outcome
11.
Allergy ; 61(5): 589-97, 2006 May.
Article in English | MEDLINE | ID: mdl-16629789

ABSTRACT

BACKGROUND: The effector function of eosinophils involves their release of toxic granule proteins, reactive oxygen species, cytokines, and lipid mediators. Murine studies have demonstrated that eosinophils can also enhance T cell function. Whether human eosinophils, in particular, airway eosinophils, have similar immunoregulatory activity has not been fully investigated. The aim of this study was to determine whether human blood and airway eosinophils can contribute to Th1 and Th2 cytokine generation from CD4+ T cells stimulated with superantigen. METHODS: Eosinophils were obtained from blood or bronchoalveolar lavage fluid 48 h after segmental allergen bronchoprovocation. Purified eosinophils were co-cultured with autologous CD4+ blood T cells in the presence of staphylococcal enterotoxin B (SEB). Cytokine levels in the supernatant fluid were determined by enzyme-linked immunosorbent assay (ELISA). Eosinophil expression of major histocompatibility complex (MHC) class II and co-stimulatory molecules was assessed by flow cytometry before culture, 24 h after granulocyte-macrophage colony-stimulating factor (GM-CSF) stimulation, and 24 h after co-culture with CD4+ T cells and SEB. RESULTS: Interleukin (IL)-5, IL-13, and interferon (IFN)-gamma generation increased when CD4+ T cells were co-cultured with either blood or airway eosinophils in the presence of SEB. The ability of eosinophils to enhance cytokine generation was independent of their source (blood vs airway), activation by GM-CSF, or detectable expression of human leukocyte antigen (HLA)-DR, CD80, or CD86. CONCLUSION: Our data demonstrate that SEB-induced generation of Th1 and Th2 cytokines is increased in the presence of human blood and airway eosinophils. Thus, eosinophils can have an immunoregulatory function in pathogen-associated allergic diseases such as atopic dermatitis, chronic sinusitis, and asthma exacerbations.


Subject(s)
Bronchoalveolar Lavage Fluid , Cytokines/metabolism , Eosinophils/physiology , Th1 Cells/metabolism , Th2 Cells/metabolism , Blood Cells/physiology , CD4-Positive T-Lymphocytes/metabolism , Cells, Cultured , Enterotoxins/metabolism , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Granulocyte-Macrophage Colony-Stimulating Factor/metabolism , Humans , In Vitro Techniques , Interferons/metabolism , Interleukin-5/metabolism , Th1 Cells/immunology , Th2 Cells/immunology
12.
Clin Exp Allergy ; 34(2): 227-33, 2004 Feb.
Article in English | MEDLINE | ID: mdl-14987302

ABSTRACT

BACKGROUND: Nocturnal enhancement of airway inflammation has been demonstrated in patients with asthma who have a significant drop in pulmonary function at night. OBJECTIVE: To investigate the circadian changes in airway inflammation and their relationship with variations in pulmonary function in subjects with mild atopic asthma. METHODS: Twelve asthma subjects were admitted to the hospital for two separate 24-h visits. Bronchoalveolar lavage (BAL) was performed at 04:00 hours during one visit, and at 16:00 hours during another visit. BAL cells were analysed for lymphocyte phenotype and the capacity to secrete cytokines following ex vivo stimulation with phytohaemagglutinin (PHA). RESULTS: The numbers of BAL lymphocytes and the percentage of CD4+ T cells were higher at 04:00 hours compared with 16:00 hours. At 04:00 hours, the forced expiratory volume in 1 s (FEV1) was inversely correlated with BAL lymphocytes and CD4+ cells. PHA-induced generation of IL-5 by BAL cells correlated with BAL eosinophils and CD4+ cells. Moreover, there was a linear relationship between the relative change (16:00-04:00 hours) in IL-5 and circadian variation in FEV1. CONCLUSIONS: These data suggest that the circadian variation in lung function in asthma is associated with increased airway CD4+ lymphocyte numbers and their capacity to generate IL-5. Furthermore, in mild asthma, these circadian changes appear to fall into a continuous range, suggesting that day/night variations in airway inflammation and lung function occur on a continuum, rather than as an all-or-none phenomenon.


Subject(s)
Asthma/immunology , Asthma/physiopathology , Circadian Rhythm , Lung/immunology , Lung/physiopathology , Adult , Bronchoalveolar Lavage Fluid/immunology , CD4-Positive T-Lymphocytes/immunology , Eosinophil-Derived Neurotoxin , Female , Forced Expiratory Volume , Humans , Interleukin-5/analysis , Lymphocyte Count , Macrophages, Alveolar/immunology , Male , Neutrophils/immunology , Phytohemagglutinins/pharmacology , Ribonucleases/analysis , Statistics, Nonparametric
13.
Respir Med ; 94 Suppl F: S3-8, 2000 Oct.
Article in English | MEDLINE | ID: mdl-11059961

ABSTRACT

Bronchial biopsy provides valuable information about the inflammatory processes in lung tissue, but optimal results are only achieved if the design of intervention studies is sufficiently rigorous. The parallel-group design has merit, but the cross-over design is statistically superior, providing the wash-out period is effective. Heterogeneity of contributing pathologies in asthma patients results in large inter-patient variability which must be controlled for, for example by using strict inclusion criteria, which should ideally relate to the specific inflammatory marker being studied. The inclusion of a placebo group helps to quantify sample variability. The study must have sufficient statistical power to detect inter-group differences for each variable; appropriate adjustments should be made when multiple tests are used. Studies with larger patient numbers are best performed using a multi-centre design, with one centre analysing all tissue samples to reduce variability. Study duration depends on the type of investigation, but should ideally be short. Longer studies are necessary to evaluate chronic changes such as tissue remodelling. Changes in clinical status and cellular events may follow different time courses after intervention. Biopsy measurements are less reproducible than physiological tests, and diurnal variation in the number and function of inflammatory cells can further complicate measurement. The timing of clinical trial assessments needs to allow for these idiosyncrasies. Finally, a balance must be maintained between the risk, albeit small, and the benefit of performing bronchial biopsies.


Subject(s)
Biopsy , Bronchi/pathology , Bronchial Diseases/pathology , Clinical Trials as Topic/methods , Research Design/standards , Bronchial Diseases/surgery , Cross-Over Studies , Humans , Patient Selection , Reproducibility of Results
14.
Am J Respir Crit Care Med ; 162(3 Pt 1): 883-90, 2000 Sep.
Article in English | MEDLINE | ID: mdl-10988100

ABSTRACT

To define the mechanisms by which inhaled glucocorticosteroid regulates allergen-induced airway inflammation, a double-blind, placebo-controlled, cross-over study with inhaled budesonide was conducted in 14 subjects with allergic asthma. After baseline bronchoscopy and bronchoalveolar lavage (BAL), subjects were randomized to budesonide (400 microgram, twice daily) or placebo treatment for 4 wk. At the end of each treatment phase, whole-lung allergen inhalation challenge was performed, followed by BAL 48 h later. Budesonide treatment improved the FEV(1), attenuated both the immediate- and late-phase response to allergen, and prevented the increase in bronchial hyperresponsiveness after allergen challenge. Budesonide treatment also decreased allergen-induced airway eosinophilia. To determine the effects of budesonide on airway cell function, BAL cells were stimulated ex vivo with the T cell mitogen PHA, and cytokine generation was measured by ELISA. Budesonide decreased ex vivo generation of IL-5 and IFN-gamma by BAL cells. Ex vivo IL-5 production was significantly correlated with the number of airway eosinophils (r(s) = 0.61), and levels of eosinophil-derived neurotoxin (EDN) (r(s) = 0.57) and IL-5 (r(s) = 0.52) in BAL fluid. Moreover, PHA-induced IL-5 generation correlated with FEV(1) fall during the late-phase response to allergen (r(s) = 0.60). Budesonide decreased circulating eosinophils and serum levels of IL-5, but did not reduce IL-5 generation by peripheral blood mononuclear cells. The reduction in circulating eosinophils correlated with the decrease in levels of EDN (r(s) = 0.61) in BAL fluid and late response to inhaled allergen (r(s) = 0.51). These findings suggest that long-term treatment with inhaled budesonide reduces airway cell generation of cytokines, specifically IL-5, which then decreases circulating eosinophils and their availability for recruitment to the airway after allergen exposure.


Subject(s)
Allergens , Anti-Inflammatory Agents/administration & dosage , Asthma/drug therapy , Budesonide/administration & dosage , Respiratory Hypersensitivity/drug therapy , Systemic Inflammatory Response Syndrome/drug therapy , Administration, Inhalation , Adult , Allergens/immunology , Anti-Inflammatory Agents/adverse effects , Asthma/immunology , Budesonide/adverse effects , Cross-Over Studies , Double-Blind Method , Female , Forced Expiratory Volume/drug effects , Humans , Inflammation Mediators/metabolism , Male , Respiratory Hypersensitivity/immunology , Systemic Inflammatory Response Syndrome/immunology
15.
Am J Respir Crit Care Med ; 162(3 Pt 1): 1157-61, 2000 Sep.
Article in English | MEDLINE | ID: mdl-10988146

ABSTRACT

Persistent asthma is associated with airway inflammation, tissue damage, and deposition of extracellular matrix (ECM) proteins, which may be mediated, in part, through release of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinase-1 (TIMP-1). To investigate the role of allergen in the induction of MMP-9 and TIMP-1, bronchoscopy and segmental bronchoprovocation (SBP) with saline (SAL) and antigen (AG) were performed in 17 allergic subjects. Bronchoalveolar lavage (BAL) was done 5 min and 48 h after challenge and concentrations of MMP-9 and TIMP-1 in BAL fluid (BALF) were measured by ELISA. Forty-eight hours after AG challenge, concentrations of MMP-9 and TIMP-1 were increased in the airway, but not in serum. Zymography demonstrated that MMP-9 was the predominant metalloproteinase and was present in a latent proform. MMP-9 immunoreactivity was associated primarily with neutrophils, and concentrations of MMP-9 in BALF correlated with airway neutrophils and, to a lesser extent, with alveolar macrophages. These data suggest that AG challenge leads to generation of factors, including MMP-9, that may be associated with the initiation of bronchial injury, which may then lead to remodeling in asthma.


Subject(s)
Asthma/physiopathology , Bronchoalveolar Lavage Fluid/chemistry , Matrix Metalloproteinase 9/analysis , Respiratory Hypersensitivity/physiopathology , Adult , Allergens , Bronchial Provocation Tests , Enzyme-Linked Immunosorbent Assay , Female , Humans , Male , Matrix Metalloproteinase 9/physiology , Tissue Inhibitor of Metalloproteinase-1/analysis , Tissue Inhibitor of Metalloproteinase-1/physiology
16.
Ann Allergy Asthma Immunol ; 85(2): 115-20, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10982218

ABSTRACT

BACKGROUND: Although preferential expression of the Th2 cytokines, IL-4 and IL-5, has been described in atopic asthma, the role of IFN-gamma and IL-10 are less clear. OBJECTIVE: To determine the cytokine pattern of T cell mitogen-induced peripheral blood mononuclear cells obtained from atopic asthmatic (AA) subjects. METHODS: Peripheral blood mononuclear cells obtained from AA (n = 24), allergic rhinitis (AR) (n = 9), and normals (NL) (n = 9) were stimulated with phytohemagglutinin (PHA) and the generation of IL-4, IL-5, IFN-gamma, IL-10, and GM-CSF was quantified using ELISA. RESULTS: Compared with NL subjects, peripheral blood mononuclear cells from the atopic groups had increased generation of both IL-5 (AA, P = .001 and AR, P = .024) and IFN-gamma (AA, P = .037 and AR, P = .048) and decreased generation of IL-10 (AA, P = .038 and AR, P = .036). The absolute levels of cytokines did not differ between the two atopic groups; however, the ratio of IL-5/IL-10 was significantly higher in AA (P < .05), but not in AR when compared with NL subjects. CONCLUSION: The concomitant increase in the generation of IL-5 and IFN-gamma, with a decrease in IL-10 in the atopic groups suggests that in, at least a subset of these patients, there is potential expression of both Th2- and Th1-type cytokines. Furthermore, the increased IL-5 to IL-10 ratio could represent a key feature that distinguishes atopic asthmatic from non-asthmatic atopic subjects.


Subject(s)
Asthma/blood , Cytokines/biosynthesis , Hypersensitivity, Immediate/blood , Leukocytes, Mononuclear/metabolism , Adolescent , Adult , Granulocyte-Macrophage Colony-Stimulating Factor/biosynthesis , Humans , Interferon-gamma/biosynthesis , Interleukin-10/biosynthesis , Interleukin-4/biosynthesis , Interleukin-5/biosynthesis , Th1 Cells/metabolism , Th2 Cells/metabolism
17.
J Allergy Clin Immunol ; 105(6 Pt 1): 1169-77, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10856152

ABSTRACT

BACKGROUND: Viral respiratory tract infections are the most frequent cause of asthma exacerbations. Of the respiratory viruses associated with these exacerbations, rhinovirus (RV) is the most common. It is proposed that these RV infections may enhance airway inflammation and thus provoke asthma. OBJECTIVE: It is our hypothesis that RV infections generate nasal proinflammatory mediators that are associated with an initial increase in circulating leukocytes and may contribute to later development of neutrophilic airway inflammation. METHODS: To evaluate this hypothesis, subjects with a history of allergic asthma were experimentally inoculated with strain 16 RV (RV16). The effect of this experimental infection was evaluated on circulating leukocytes, nasal-derived mediators, and markers of bronchial inflammation that were obtained by bronchoscopy and lavage. RESULTS: RV16 inoculation was associated with an initial increase in circulating neutrophils. Paralleling these acute changes in circulating neutrophils was an increase in nasal concentrations of IL-8 and granulocyte-colony-stimulating factor (G-CSF). The RV16-associated changes in circulating and nasal G-CSF correlated with increases in peripheral blood neutrophils (r(s) = 0.874, P <. 001 and r(s) = 0.898, P <.001, respectively). Bronchial lavage samples showed no increase in neutrophils 48 hours after RV16 inoculation; however, 96 hours after RV inoculation there was a significant increase in bronchial neutrophils compared with preinoculation values. CONCLUSIONS: These results suggest that the production of nasal mediators associated with the RV infection, particularly G-CSF, may be important to the eventual development of neutrophilic bronchial inflammation and thus contribute to asthma exacerbations.


Subject(s)
Bronchoalveolar Lavage Fluid/cytology , Neutrophils/virology , Picornaviridae Infections/physiopathology , Adult , Asthma/physiopathology , Bronchoalveolar Lavage Fluid/chemistry , Bronchoscopy , Female , Forced Expiratory Volume/physiology , Granulocyte Colony-Stimulating Factor/blood , Humans , Male , Peroxidase/analysis , Rhinovirus
18.
Chronobiol Int ; 16(5): 631-9, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10513886

ABSTRACT

In a majority of patients, exacerbations of asthma occur more frequently during the night than day. Many hypotheses have been proposed to explain such variation in asthma. The airways of asthmatic persons are characterized by an abnormal degree of inflammation and bronchial hyperresponsiveness to both nonspecific and specific challenges. Studies of both children and adults with asthma document marked circadian rhythmicity in the response of airways to bronchial challenges with histamine, methacholine, acetylcholine, saline, and house dust mite. Taken together, the findings of these investigations indicate that the hyperreactivity of airways to these agents is more profound and prolonged following evening and overnight tests compared to tests conducted in the midday and afternoon. The temporal pattern in bronchial response to the hyperventilation of cold dry air is different. The hyperresponsiveness of airways to this challenge is greatest in the afternoon. The amplitude of the circadian rhythm in airway hyperreactivity seems to be correlated to the amplitude of the circadian rhythm of pulmonary function; the greater the day-night difference in bronchial reactivity is, the greater is the day-night difference in flow rates.


Subject(s)
Asthma/physiopathology , Bronchial Hyperreactivity , Circadian Rhythm , Adult , Allergens/administration & dosage , Animals , Asthma/etiology , Bronchial Provocation Tests , Child , Humans , Mites/immunology
19.
Am J Respir Crit Care Med ; 160(1): 336-41, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10390421

ABSTRACT

If pulmonary surfactant develops a dysfunction, its ability to maintain patency of narrow conducting airways diminishes, which is likely to cause an increased airway resistance. We hypothesized that antigen challenge will cause inflammation in the conducting airways and that this will cause a surfactant dysfunction. Twenty atopic patients underwent bronchoalveolar lavage (BAL) 5 min and 48 h after challenge with antigen in one segment and with saline solution in another. BAL fluid (BALF) cell count, differential, and proteins were determined. Surfactant function was studied with a capillary surfactometer (CS), an instrument specifically designed to evaluate surfactant's ability to maintain patency. Eosinophils increased 80-fold 48 h after antigen challenge and total protein increased from 84 to 241 micrograms/ml (median values). BALF surfactant lost part of its ability to maintain openness of the capillary, from 68.8% to 14.0% (p < 0.05). Protein concentration negatively correlated with percent openness (r = -0.62, p = 0.005). We conclude that the antigen challenge resulted in an inflammatory reaction that caused pulmonary surfactant to lose some of its ability to maintain airway patency and speculate that surfactant dysfunction is probably an important factor contributing to increased airway obstruction in allergen-induced exacerbation of asthma.


Subject(s)
Antigens/immunology , Asthma/immunology , Bronchial Hyperreactivity/immunology , Pulmonary Surfactants/physiology , Respiratory Hypersensitivity/immunology , Adult , Airway Resistance/immunology , Bronchial Provocation Tests , Bronchoalveolar Lavage Fluid/chemistry , Female , Forced Expiratory Volume/physiology , Humans , Male , Vital Capacity/physiology
20.
Am J Respir Crit Care Med ; 159(2): 619-25, 1999 Feb.
Article in English | MEDLINE | ID: mdl-9927382

ABSTRACT

Fibronectin may contribute to asthma pathogenesis by recruitment and activation of inflammatory cells, and by promotion of subepithelial fibrosis. Fibronectin is produced by several types of airway cells, including epithelial cells, fibroblasts, and alveolar macrophages. To test the hypothesis that antigen-induced airway inflammation is associated with increased local generation of fibronectin, segmental bronchoprovocation (SBP) with antigen and saline was performed in 17 atopic patients. Bronchoalveolar lavage (BAL) was performed at 5 min and 48 h after segmental challenge with saline or antigen. Fibronectin concentrations in BAL fluid, measured by enzyme-linked immunosorbent assay (ELISA), increased more than 5-fold 48 h after antigen challenge (65 [47 to 110] versus 407 [240 to 697] ng/ml, median and 25 to 75% interquartiles, p < 0.05). Fibronectin concentrations 48 h after antigen challenge correlated with histamine concentrations 5 min after antigen challenge and numbers of eosinophils, neutrophils, macrophages, and total cells in BAL fluid 48 h after antigen challenge. BAL was more enriched in fibronectin 48 h after challenge than would be predicted solely from increased permeability of plasma proteins. Western blot analysis showed that fibronectin in BAL fluid was largely intact and contained the extra domain-A (ED-A) splice variant of cellular fibronectin, indicative of local production. We conclude that antigen challenge in atopic subjects causes increased production of fibronectin by airway cells and speculate that this response may contribute to airway remodeling in allergic inflammation.


Subject(s)
Antigens/adverse effects , Bronchoalveolar Lavage Fluid/chemistry , Fibronectins/biosynthesis , Respiratory Hypersensitivity/metabolism , Adult , Albumins/metabolism , Biomarkers , Blotting, Western , Bronchial Provocation Tests , Bronchoalveolar Lavage Fluid/cytology , Enzyme-Linked Immunosorbent Assay , Eosinophils/metabolism , Epithelium/metabolism , Female , Follow-Up Studies , Histamine/metabolism , Humans , Leukocyte Count , Lymphocytes/metabolism , Macrophages, Alveolar/metabolism , Male , Neutrophils/metabolism , Respiratory Function Tests , Respiratory Hypersensitivity/etiology
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