RESUMO
BACKGROUND: The severity of bronchial hyperresponsiveness (BHR) is a fundamental feature of asthma. The severity of BHR varies between asthmatics and is associated with lack of asthma control. The mechanisms underlying this trait are still unclear. This study aimed to identify genes associated with BHR severity, using a genomewide association study (GWAS) on the slope of BHR in adult asthmatics. METHODS: We performed a GWAS on BHR severity in adult asthmatics from the Dutch Asthma GWAS cohort (n = 650), adjusting for smoking and inhaled corticosteroid use, and verified results in three other cohorts. Furthermore, we performed eQTL and co-expression analyses in lung tissue. RESULTS: In the discovery cohort, one genomewide significant hit located in phosphodiesterase 4D, cAMP-specif (PDE4D) and 26 SNPs with P-values < 1*10-5 were found. None of our findings replicated in adult and childhood replication cohorts jointly. In adult cohorts separately, rs1344110 in pituitary tumour-transforming 1 interacting protein (PTTG1IP) and rs345983 in Mastermind-like 3 (MAML3) replicated nominally; minor alleles of rs345983 and rs1344110 were associated with less severe BHR and higher lung tissue gene expression. PTTG1IP showed significant co-expression with pituitary tumour-transforming 1, the binding factor of PTTG1lP, and with vimentin and E-cadherin1. MAML3 co-expressed significantly with Mastermind-like 2 (MAML2), both involved in Notch signalling. CONCLUSIONS: PTTG1IP and MAML3 are associated with BHR severity in adult asthma. The relevance of these genes is supported by the eQTL analyses and co-expression of PTTG1lP with vimentin and E-cadherin1, and MAML3 with MAML2.
Assuntos
Asma/genética , Hiper-Reatividade Brônquica/genética , Proteínas de Ligação a DNA/genética , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Proteínas de Membrana/genética , Proteínas Nucleares/genética , Fatores de Transcrição/genética , Adulto , Asma/diagnóstico , Hiper-Reatividade Brônquica/diagnóstico , Estudos de Coortes , Feminino , Expressão Gênica , Genótipo , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Testes de Função Respiratória , Índice de Gravidade de Doença , TransativadoresRESUMO
BACKGROUND: Asthma is a biologically heterogeneous disease and development of novel therapeutics requires understanding of pathophysiologic phenotypes. There is uncertainty regarding the stability of clinical characteristics and biomarkers in asthma over time. This report presents the longitudinal stability over 12 months of clinical characteristics and clinically accessible biomarkers from ADEPT. METHODS: Mild, moderate, and severe asthma subjects were assessed at 5 visits over 12 months. Assessments included patient questionnaires, spirometry, bronchodilator reversibility, fractional exhaled nitric oxide (FENO), and biomarkers measured in induced sputum. RESULTS: Mild (n = 52), moderate (n = 55), and severe (n = 51) asthma cohorts were enrolled from North America and Western Europe. For all clinical characteristics and biomarkers, group mean data showed no significant change from visit to visit. However, individual data showed considerable variability. FEV1/FVC ratio showed excellent reproducibility while pre-bronchodilator FEV1 and FVC were only moderately reproducible. Of note bronchodilator FEV1 reversibility showed low reproducibility, with the nonreversible phenotype much more reproducible than the reversible phenotype. The 7-item asthma control questionnaire (ACQ7) demonstrated moderate reproducibility for the combined asthma cohorts, but the uncontrolled asthma phenotype (ACQ7 > 1.5) was inconstant in mild and moderate asthma but stable in severe asthma. FENO demonstrated good reproducibility, with the FENO-low phenotype (FENO < 35 ppb) more stable than the FENO-high phenotype (FENO ≥ 35 ppb). Induced sputum inflammatory phenotypes showed marked variability across the 3 sputum samples taken over 6 months. CONCLUSIONS: The ADEPT cohort showed group stability, individual stability in some parameters e.g. low FEV1/FVC ratio, and low FENO, but marked individual variability in other clinical characteristics and biomarkers e.g. type-2 biomarkers over 12 months. This variability is possibly related to seasonal variations in climate and allergen exposure, medication changes and acute exacerbations. The implications for patient selection strategies based on clinical biomarkers may be considerable.
Assuntos
Asma/tratamento farmacológico , Testes de Função Respiratória/estatística & dados numéricos , Escarro/citologia , Adulto , Asma/epidemiologia , Biomarcadores , Broncodilatadores/uso terapêutico , Europa (Continente)/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , América do Norte/epidemiologia , Prevalência , Reprodutibilidade dos Testes , Fatores de Risco , Sensibilidade e Especificidade , Índice de Gravidade de Doença , Resultado do TratamentoRESUMO
BACKGROUND: Asthma is a heterogeneous disease and development of novel therapeutics requires an understanding of pathophysiologic phenotypes. The purpose of the ADEPT study was to correlate clinical features and biomarkers with molecular characteristics, by profiling asthma (NCT01274507). This report presents for the first time the study design, and characteristics of the recruited subjects. METHODS: Patients with a range of asthma severity and healthy non-atopic controls were enrolled. The asthmatic subjects were followed for 12 months. Assessments included history, patient questionnaires, spirometry, airway hyper-responsiveness to methacholine, fractional exhaled nitric oxide (FENO), and biomarkers measured in induced sputum, blood, and bronchoscopy samples. All subjects underwent sputum induction and 30 subjects/cohort had bronchoscopy. RESULTS: Mild (n = 52), moderate (n = 55), severe (n = 51) asthma cohorts and 30 healthy controls were enrolled from North America and Western Europe. Airflow obstruction, bronchodilator response and airways hyperresponsiveness increased with asthma severity, and severe asthma subjects had reduced forced vital capacity. Asthma control questionnaire-7 (ACQ7) scores worsened with asthma severity. In the asthmatics, mean values for all clinical and biomarker characteristics were stable over 12 months although individual variability was evident. FENO and blood eosinophils did not differ by asthma severity. Induced sputum eosinophils but not neutrophils were lower in mild compared to the moderate and severe asthma cohorts. CONCLUSIONS: The ADEPT study successfully enrolled asthmatics across a spectrum of severity and non-atopic controls. Clinical characteristics were related to asthma severity and in general asthma characteristics e.g. lung function, were stable over 12 months. Use of the ADEPT data should prove useful in defining biological phenotypes to facilitate personalized therapeutic approaches.
Assuntos
Antiasmáticos/uso terapêutico , Asma/diagnóstico , Asma/tratamento farmacológico , Broncodilatadores/uso terapêutico , Pulmão/efeitos dos fármacos , Medicina de Precisão , Adolescente , Adulto , Idoso , Asma/epidemiologia , Asma/metabolismo , Asma/fisiopatologia , Biomarcadores/metabolismo , Broncoconstrição/efeitos dos fármacos , Canadá/epidemiologia , Estudos de Casos e Controles , Europa (Continente)/epidemiologia , Feminino , Humanos , Estudos Longitudinais , Pulmão/metabolismo , Pulmão/fisiopatologia , Masculino , Pessoa de Meia-Idade , Seleção de Pacientes , Fenótipo , Valor Preditivo dos Testes , Prevalência , Projetos de Pesquisa , Testes de Função Respiratória , Fatores de Risco , Índice de Gravidade de Doença , Escarro/metabolismo , Inquéritos e Questionários , Fatores de Tempo , Resultado do Tratamento , Estados Unidos/epidemiologia , Adulto JovemRESUMO
Bronchial thermoplasty is a recent endoscopic technique for the treatment of severe asthma. It is an innovative treatment whose clinical efficacy and safety are beginning to be better understood. Since this is a device-based treatment, the evaluation procedure of risks and benefits is different that for pharmaceutical products; safety aspects, regulatory requirements, study design and the assessment of the magnitude of effects may all be different. The mechanism of action and optimal patient selection need to be assessed further in rigorous clinical and scientific studies. This technique is in harmony with the development of personalised medicine in the 21st century. It should be developed further in response to the numerous challenges and needs not yet met in the management of severe asthma.
Assuntos
Asma/cirurgia , Brônquios/cirurgia , Broncoscopia/métodos , Eletrocoagulação/métodos , Adolescente , Adulto , Idoso , Asma/epidemiologia , Broncoscopia/efeitos adversos , Ablação por Cateter/efeitos adversos , Ablação por Cateter/métodos , Eletrocoagulação/efeitos adversos , Humanos , Pessoa de Meia-Idade , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologia , Índice de Gravidade de Doença , Adulto JovemRESUMO
BACKGROUND: The OX40/OX40L interaction contributes to an optimal T cell response following allergic stimuli and plays an important role in the maintenance and reactivation of memory T effector cells. OBJECTIVE: We tested whether treatment with an anti-OX40L monoclonal antibody (MAb) would inhibit allergen-induced responses in subjects with asthma. METHODS: Twenty-eight mild, atopic asthmatic subjects were recruited for a double-blind, randomized, placebo-controlled, parallel-group trial (ClinicalTrials.gov identifier NCT00983658) to compare blockade of OX40L using a humanized anti-OX40L MAb to placebo-administered intravenously in 4 doses over 3 months. Allergen inhalation challenges were carried out 56 and 113 days after the first dose of study drug. The primary outcome variable was the late-phase asthmatic response. Other outcomes included the early-phase asthmatic response, airway hyperresponsiveness, serum IgE levels, blood and sputum eosinophils, safety and tolerability. RESULTS: Treatment with anti-OX40L MAb did not attenuate the early- or late-phase asthmatic responses at days 56 or 113 compared with placebo. In the anti-OX40L MAb treatment group, total IgE was reduced 17% from pre-dosing levels, and sputum eosinophils decreased 75% by day 113 (both P = 0.04). There was no effect of anti-OX40L MAb on airway hyperresponsiveness or blood eosinophils. The frequency of AEs was similar in both groups. CONCLUSION AND CLINICAL RELEVANCE: Pharmacological activity of anti-OX40L MAb was observed by decreases in serum total IgE and airway eosinophils at 16 weeks post-dosing, but there was no effect on allergen-induced airway responses. It is possible that the treatment duration or dose of antibody was insufficient to impact the airway responses.
Assuntos
Alérgenos/imunologia , Antiasmáticos/uso terapêutico , Anticorpos Monoclonais/uso terapêutico , Asma/tratamento farmacológico , Asma/imunologia , Ligante de CD40/antagonistas & inibidores , Adulto , Antiasmáticos/efeitos adversos , Antiasmáticos/farmacologia , Anticorpos Monoclonais/efeitos adversos , Anticorpos Monoclonais/farmacologia , Asma/metabolismo , Antígenos CD40/metabolismo , Ligante de CD40/metabolismo , Células Dendríticas/imunologia , Eosinófilos , Feminino , Volume Expiratório Forçado/efeitos dos fármacos , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Transdução de Sinais/efeitos dos fármacos , Linfócitos T/imunologia , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
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.
Assuntos
Remodelação das Vias Aéreas , Asma/imunologia , Asma/patologia , Eosinofilia/patologia , Adulto , Asma/metabolismo , Estudos de Casos e Controles , Feminino , Células Caliciformes/patologia , Humanos , Hiperplasia , Masculino , Pessoa de Meia-Idade , Mucinas/metabolismo , Músculo Liso/metabolismo , Músculo Liso/patologia , Mucosa Respiratória/imunologia , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologia , Adulto JovemRESUMO
AIM: The limited potential of exercise to induce weight loss could be partly due to the overestimation of the energy cost of exercise. The objectives of this study were twofold: 1) to investigate whether men and women are able to accurately estimate exercise energy expenditure (EE); and 2) to determine whether they have the ability to accurately compensate for the EE of exercise during a buffet-type meal. METHODS: Sixteen (8 men, 8 women) moderately active (VO2 peak=45.4±7.7 mL.kg-1.min-1), normal weight (BMI=22.8±3.3 kg/m2) individuals, aged 20-35 years, were studied. They were blinded to two randomly assigned experimental conditions: a 200 and a 300 kcal (measured by indirect calorimetry) exercise sessions that were performed on a treadmill at the same intensity (50% of VO2 peak). At the end of each exercise session individuals were asked to estimate EE of the exercise sessions and to then eat the caloric equivalent of their estimated exercise EE from a buffet-type meal. RESULTS: Estimated EE was higher than measured EE for both the 200 kcal (825.0±1061.8 vs. 200.1±0.7 kcal, P<0.05) and 300 kcal (896.9±952.4 vs. 300.2±0.7 kcal, P<0.05) sessions. Further, post-exercise energy intake was higher than measured EE for the 200 kcal (556.8±204.4 vs. 200.1±0.7 kcal, P<0.001) and the 300 kcal (607.2±166.5 vs. 300.2±0.7 kcal, P<0.001) sessions. Although post-exercise energy intake was lower than estimated EE, no significant differences were noted. CONCLUSION: These results suggest that normal weight individuals overestimate EE during exercise by 3-4 folds. Further, when asked to precisely compensate for exercise EE with food intake, the resulting energy intake is still 2 to 3 folds greater than the measured EE of exercise.
Assuntos
Metabolismo Energético , Exercício Físico , Adulto , Calorimetria Indireta , Estudos Cross-Over , Ingestão de Energia , Teste de Esforço , Feminino , Humanos , Masculino , Método Simples-Cego , Adulto JovemRESUMO
BACKGROUND: Bronchial epithelium is considered a key player in coordinating airway wall remodelling. The function of epithelial cells can be modulated by the underlying fibroblasts through autocrine and paracrine mechanisms. OBJECTIVE: To investigate the effect of phenotypic changes in bronchial fibroblasts from asthmatic subjects on epithelial cell proliferation. METHODS: Epithelial cells and fibroblasts derived from bronchial biopsies of asthmatic and healthy controls were cultured in an engineered model. Proliferation was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium-bromid (MTT). Epidermal growth factor receptor (EGFR), cyclin-dependent kinase inhibitors p21 and p27 were measured by western blots. Total and active forms of transforming growth factor (TGF)-ß1 were measured using ELISA and bioassay. TGF-ß was inhibited using a recombinant TGF-ß soluble receptor II protein. RESULTS: Proliferation of epithelial cells from asthmatics (AE) is increased when cells were cultured with fibroblasts from normal controls (NF). Fibroblasts from asthmatics (AF) significantly decreased the proliferation of epithelial cells from healthy subjects (NE). Activation of p21, p27, EGFR and TGF-ß1 reflects the proliferation data by decreasing in AE cultured with NF and increasing in NE cultured with AF. Neutralization of TGF-ß increased proliferation of epithelial cells cultured in the asthmatic model. CONCLUSION: Fibroblasts from asthmatic subjects regulate epithelial cell prolifearation, and TGF-ß signalling may represent one of the pathway involved in these interactions.
Assuntos
Asma/metabolismo , Células Epiteliais/metabolismo , Fibroblastos/metabolismo , Mucosa Respiratória/metabolismo , Transdução de Sinais/fisiologia , Asma/imunologia , Asma/patologia , Western Blotting , Brônquios/imunologia , Brônquios/metabolismo , Brônquios/patologia , Comunicação Celular/fisiologia , Proliferação de Células , Células Cultivadas , Técnicas de Cocultura , Proteínas Inibidoras de Quinase Dependente de Ciclina/biossíntese , Proteínas Inibidoras de Quinase Dependente de Ciclina/imunologia , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/imunologia , Células Epiteliais/ultraestrutura , Receptores ErbB/metabolismo , Fibroblastos/imunologia , Fibroblastos/ultraestrutura , Humanos , Microscopia Eletrônica de Varredura , Mucosa Respiratória/imunologia , Mucosa Respiratória/patologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Engenharia Tecidual , Fator de Crescimento Transformador beta/biossíntese , Fator de Crescimento Transformador beta/imunologiaRESUMO
Normalisation of eosinophil counts in sputum of asthmatic patients reduces eosinophilic exacerbations. However, the effect of this strategy on airway remodelling remains to be determined. We compared bronchial inflammation and collagen deposition after 2 yrs of treatment guided by either sputum eosinophils (sputum strategy, SS) or by clinical criteria (clinical strategy, CS). As a pilot study, 20 mild asthmatic patients were randomly assigned to CS or SS strategies. Bronchial biopsies were obtained when minimum treatment needed to maintain control was identified and this was continued for 2 yrs. Biopsies were immunostained for inflammatory cells, mucin 5A (MUC5A) and collagen. The mean dose of inhaled corticosteroids at the start and end of the study was similar in both SS and CS groups. Forced expiratory volume in 1 s increased in both groups at the study end. In SS, mucosal lymphocyte and eosinophil counts, but not neutrophils, were reduced at the end of the study. In CS, only activated eosinophil and neutrophil counts decreased. MUC5A staining decreased in SS but not CS. No change in collagen deposition underneath the basement membrane was observed in either strategy. Treatment strategies that normalise sputum eosinophils also reduce mucosal inflammatory cells and MUC5A expression, but do not change subepithelial collagen deposition in mild to moderate asthma.
Assuntos
Remodelação das Vias Aéreas , Asma/imunologia , Bronquite/imunologia , Eosinófilos , Escarro/citologia , Adulto , Asma/patologia , Biópsia , Bronquite/patologia , Contagem de Células , Feminino , Humanos , Masculino , Projetos PilotoRESUMO
BACKGROUND: Although exposure to tobacco smoke has been associated with increased morbidity and mortality, cigarette smoking is still common in the asthmatic population. Induced sputum neutrophilia has been observed in asthmatic smokers, but the effects of regular smoking on their bronchial mucosa morphology remain to be defined. This study documents the inflammatory and remodelling features in bronchial biopsies of smoking compared with non-smoking asthmatics. METHODS: We analysed bronchial biopsies from 24 steroid-naïve young subjects with mild asthma: 12 non-smoking and 12 currently smoking subjects. In addition to airway morphology assessment, inflammation and remodelling were analysed by immunohistochemistry using antibodies against CD3, CD68, major basic protein, neutrophil elastase, and tryptase. Expression of the cytokines IL-4, IL-5, IL-8, IFN-gamma, transforming growth factor-beta, and TNF was determined by in situ hybridization. RESULTS: Compared with non-smoking asthmatic subjects, smoking asthmatics' bronchial mucosa showed squamous cell metaplasia, in addition to increased expression of subepithelial neutrophil elastase, IFN-gamma, and intraepithelial IL-8. CONCLUSIONS: Smoking status modifies morphological and inflammatory processes in young subjects with mild asthma. The changes may possibly affect asthma treatment responses and clinical outcomes.
Assuntos
Asma/imunologia , Bronquite/imunologia , Citocinas/imunologia , Mucosa Respiratória/imunologia , Fumar/efeitos adversos , Adulto , Asma/patologia , Bronquite/patologia , Citocinas/biossíntese , Células Epiteliais/citologia , Células Epiteliais/imunologia , Células Epiteliais/patologia , Feminino , Humanos , Masculino , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologia , Fumar/imunologiaRESUMO
The aim of the present study was to compare the effectiveness, safety and health economics of budesonide/formoterol maintenance and a novel reliever therapy with conventional best practice in patients with persistent asthma in Canada. After 2 weeks of usual therapy, 1,538 patients were randomised for 6 months to open-label budesonide/formoterol maintenance and reliever therapy 160/4.5 microg twice daily and as needed, or to guideline-based conventional best practice. Severe asthma exacerbations, reliever medication use and total inhaled corticosteroid dose were analysed in all patients and airway inflammation was assessed in a sub-study of 115 patients. No differences were seen in time to first severe exacerbation and severe asthma exacerbation rate. There were numerically fewer emergency room visits or hospitalisations with budesonide/formoterol maintenance and reliever therapy (4.4 versus 7.5 events per 100 patients x yr(-1), 41% reduction); however, this did not reach statistical significance. Mean total inhaled corticosteroid dose, reliever use, asthma medication costs and total annual costs per patient were all significantly lower with budesonide/formoterol maintenance and reliever therapy. Mean sputum eosinophil cell counts remained in the range for controlled inflammation in both groups. In conclusion, budesonide/formoterol maintenance and reliever therapy achieved similar or improved clinical control compared with conventional best practice, with significantly lower total inhaled corticosteroid dose and lower cost, while maintaining similar control of eosinophilic inflammation.
Assuntos
Anti-Inflamatórios/uso terapêutico , Asma/tratamento farmacológico , Broncodilatadores/uso terapêutico , Budesonida/uso terapêutico , Etanolaminas/uso terapêutico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anti-Inflamatórios/efeitos adversos , Anti-Inflamatórios/economia , Asma/economia , Broncodilatadores/efeitos adversos , Broncodilatadores/economia , Budesonida/efeitos adversos , Budesonida/economia , Criança , Análise Custo-Benefício , Quimioterapia Combinada , Etanolaminas/efeitos adversos , Etanolaminas/economia , Feminino , Fumarato de Formoterol , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do TratamentoRESUMO
CX3CR1, a fractalkine receptor, mediates cell-adhesive and migratory functions in inflammation. Based on CX3CR1 expression observed in bronchial tissues of asthmatic subjects, we hypothesized that genetic variation at this locus may affect susceptibility to asthma. We carried out an association study and a haplotypic analysis with selected polymorphisms of the CX3CR1 in a familial asthmatic sample from a founder population. Genetic analyses performed by FBAT software showed five CX3CR1 single nucleotide polymorphisms (rs938203, rs2669849, rs1050592, T280M and V249I) with significant associations between their common alleles and asthma (P<0.004) in a dominant model. A haplotype formed with common alleles of rs1050592, T280M and V249I is also overtransmitted in asthmatic subjects (P=0.005) under a dominant model. The associations of V249I and rs2669849 have been validated in an independent case-control sample. For V249I, odds ratios (OR) are 2.16 (common homozygous) and 2.11 (heterozygous) in dominant model (P=0.031). For rs2669849, OR are 2.75 (common homozygous) and 1.86 (heterozygous) in additive model (P=0.007) and dominant model (P=0.059). These results suggest an asthma protective effect of the minor alleles in healthy control carriers. Further functional studies of CX3CR1 are needed to document its role in the pathophysiology of asthma.
Assuntos
Asma/genética , Predisposição Genética para Doença , Variação Genética , Receptores de Quimiocinas/genética , Adolescente , Adulto , Receptor 1 de Quimiocina CX3C , Feminino , Componentes do Gene , Haplótipos/genética , Humanos , Desequilíbrio de Ligação , Pulmão/metabolismo , Masculino , Pessoa de Meia-Idade , Razão de Chances , Polimorfismo de Nucleotídeo Único/genética , Quebeque , Receptores de Quimiocinas/metabolismoRESUMO
One important goal of asthma treatment is to reduce exacerbations. The current authors investigated if the use of sputum cell counts to guide treatment would achieve this goal. A total of 117 adults with asthma were entered into a multicentre, randomised, parallel group-effectiveness study for two treatment strategies over a 2-yr period. In one strategy (the clinical strategy: CS) treatment was based on symptoms and spirometry. In the other (the sputum strategy: SS) sputum cell counts were used to guide corticosteroid therapy to keep eosinophils
Assuntos
Asma/diagnóstico , Asma/tratamento farmacológico , Espirometria , Escarro/citologia , Adulto , Contagem de Células , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: IL-4 is a key factor for T helper type 2 (Th2) differentiation and Ig class switching to IgE and IgG(4) during the development of immune responses. IL-4 is produced by T cells, mast cells, basophils, and eosinophils. However, there is also evidence suggesting that rat alveolar macrophages (AMs) produce IL-4. OBJECTIVE: Given the importance of AMs and Th2-related diseases in the lung, we investigated the production of IL-4 by human AMs. METHODS: Human AMs were isolated from bronchoalveolar lavage, purified, and IL-4 production was investigated at mRNA and protein levels using real-time PCR, flow cytometry, immunocytochemistry, and ELISA. The presence of IL-4 was investigated in subjects with asthma or asymptomatic airway hyper-responsiveness, and in normal non-smokers. RESULTS: IL-4 and IL-4delta2 (a splice variant found in other IL-4 producing cells) mRNAs were found in all these subjects, but IL-4 expression could not be correlated with a particular disease. Protein production was verified by immunocytochemistry and flow cytometry analysis demonstrating, respectively, up to 69% and 59% positive AMs, regardless of the subject condition. Furthermore, phorbol-12-myristate-13-acetate and calcium ionophore stimulated the release of IL-4 after 48 h treatment in the presence of anti-IL-4 receptor antibody. CONCLUSION: Our results show for the first time that IL-4 and IL-4delta2 mRNA are expressed and IL-4 protein produced and released by human AMs, suggesting a contribution of these cells in the modulation of Th2 immune response.
Assuntos
Interleucina-4/metabolismo , Macrófagos/metabolismo , Alvéolos Pulmonares/metabolismo , Líquido da Lavagem Broncoalveolar/citologia , Células Cultivadas , Citometria de Fluxo/métodos , Humanos , Imuno-Histoquímica/métodos , Interleucina-4/análise , Interleucina-4/biossíntese , Monócitos/imunologia , Isoformas de Proteínas/análise , Isoformas de Proteínas/metabolismo , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodosRESUMO
INTRODUCTION: Hypersensitivity pneumonitis (HP) is a pulmonary disease with symptoms of dyspnoea and cough resulting from the inhalation of an antigen to which the patient has been previously sensitized. STATE OF ART: Acute and subacute HP represent the most active forms of the disease which may become chronic while remaining progressive. HP may also evolve to end-stage lung disease. Clinical symptoms and signs tend to be non-specific and the diagnosis of HP often relies on the clinical context. The immune response is initiated when the alveolar macrophage phagocytoses the antigen, provoking the expansion of lymphocytes T and B that reach the pulmonary parenchyma through the systemic circulation. This reaction is amplified by the expression of a number of inflammatory mediators. PERSPECTIVE AND CONCLUSION: This article summarizes our current understanding of the diagnostic approach and immunological mechanisms related to HP.
Assuntos
Alveolite Alérgica Extrínseca/imunologia , Alveolite Alérgica Extrínseca/diagnóstico , Alveolite Alérgica Extrínseca/metabolismo , Anticorpos/metabolismo , Líquido da Lavagem Broncoalveolar/citologia , Proteínas da Matriz Extracelular/metabolismo , Humanos , Mediadores da Inflamação/metabolismo , Linfócitos/patologia , Macrófagos Alveolares/patologia , Neutrófilos/patologia , Inibidores de Proteases/metabolismo , Surfactantes Pulmonares/metabolismoRESUMO
We have reported morphological and functional features of cells isolated from human bronchial biopsies. Both epithelial and fibroblastic cells were isolated from the same biopsies using collagenase. A few models have been established to study normal bronchial response to various agents and to understand the mechanisms responsible for some disorders, such as asthma. We produced three-dimensional bronchial equivalents in culture, using human epithelial and fibroblastic cells. We previously showed that peripheral anchorage can prevent the dramatic collagen contraction in gels seeded with fibroblasts when properly adapted to the size and type of cultured tissues. Our bilayered bronchial constructs were anchored and cultured under submerged conditions and at the air-liquid interface. Three culture media were compared. Serum-free medium supplemented with retinoic acid (5 x 10(-8) M) was found to be the best for maintenance of bronchial cell properties in the reconstructed bronchial tissue. Immunohistological and ultrastructural analyses showed that these equivalents present good structural organization, allowing ciliogenesis to occur in culture. Moreover, human bronchial goblet cells could differentiate and secrete mucus with culture time. Laminin, a major constituent of the basement membrane and basal cells, was also detected at the mesenchymoepithelial interface. Such models will be useful for studying human bronchial properties in vitro.
Assuntos
Brônquios/citologia , Fibroblastos/fisiologia , Mucosa Respiratória/fisiologia , Engenharia Tecidual/métodos , Cílios/fisiologia , Meios de Cultura , Gelatinases , Humanos , Imuno-Histoquímica , Laminina/metabolismo , Microscopia Eletrônica de Varredura , Mucosa Respiratória/ultraestrutura , Tretinoína/fisiologiaRESUMO
Bronchial biopsies are currently used to study the pathophysiology of airway diseases, and comparisons are often made with biopsies from healthy volunteers. It is therefore important to evaluate the variability in each parameter analyzed in bronchial biopsies of healthy volunteers in order to be able to discriminate significant changes. We analyzed bronchial biopsies of 31 nonsmoking, nonatopic healthy subjects who volunteered as normal controls for studies on pathophysiology of asthma. Mean % epithelial desquamation was 23.7% of observed total epithelial length. No subepithelial fibrosis was observed. Inflammatory cell counts (/mm(2) connective tissue surface) were variable among subjects but not different between small (Assuntos
Brônquios/citologia
, Brônquios/patologia
, Adolescente
, Adulto
, Animais
, Asma/diagnóstico
, Asma/imunologia
, Asma/fisiopatologia
, Biópsia
, Brônquios/efeitos dos fármacos
, Testes de Provocação Brônquica
, Broncoscopia
, Contagem de Células
, Células Epiteliais/imunologia
, Células Epiteliais/metabolismo
, Feminino
, Humanos
, Imuno-Histoquímica
, Masculino
, Pessoa de Meia-Idade
, Variações Dependentes do Observador
, Valor Preditivo dos Testes
, Valores de Referência
, Testes de Função Respiratória
, Testes Cutâneos
, Fumar/imunologia
, Fumar/fisiopatologia
RESUMO
Migration from blood to tissue modulates eosinophil function, possibly through interactions with endothelial cells. The effects of contact with and migration through endothelial cells on eosinophil expression of surface markers and release of leukotriene C4 were evaluated. A small proportion (2.6%) of eosinophils spontaneously migrated through endothelial cell monolayers. Activation of endothelial cells by interleukin (IL)-4 or IL-1beta slightly increased this migration (to 12.4%), which became much greater when a chemoattractant was placed in the lower chamber (84.3%). However, the chemotactic effect was downregulated by pretreating endothelial cells with interferon gamma (IFN-gamma; 63.1%). At baseline, 5% of eosinophils expressed CD69; this increased to 30.7% in culture on untreated endothelial cells and to 50.9% on IL-1beta-pretreated endothelial cells. This effect was mediated through intercellular adhesion molecule-1/CD11b interaction. Eosinophil migration through endothelial cells further increased CD69 expression to 63.9% and also increased CD35 expression from 83.3 to 91.3%. Upon stimulation, eosinophils that had migrated through endothelial cells produced more leukotriene C4 than control cells (872.4 and 103.9 pg x mL(-1), respectively). Endothelial cell pretreatment with IL-4 or IL-1beta further increased leukotriene C4 release (1,789.1 and 2,895.1 pg x mL(-1), respectively), whereas pretreatment with IFN-gamma decreased it (293.7 pg x mL(-1)). These data show that in vitro interactions with endothelial cells upregulate eosinophil membrane receptor expression and mediator release and that these effects are differently modulated by T-helper cell type 1 and 2 cytokines. These eosinophil modulations may play an important role in asthma pathogenesis.
Assuntos
Adjuvantes Imunológicos/farmacologia , Antígenos de Superfície/análise , Asma/fisiopatologia , Movimento Celular/efeitos dos fármacos , Movimento Celular/fisiologia , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/fisiopatologia , Eosinófilos/efeitos dos fármacos , Eosinófilos/fisiologia , Interleucina-1/farmacologia , Interleucina-4/farmacologia , Leucotrieno C4/análise , Linfocinas/análise , Adolescente , Adulto , Antígenos CD/análise , Antígenos de Diferenciação de Linfócitos T/análise , Feminino , Humanos , Técnicas In Vitro , Lectinas Tipo C , Masculino , Pessoa de Meia-Idade , Receptores de Complemento 3b/análise , Linfócitos T Auxiliares-IndutoresRESUMO
BACKGROUND: Tissue eosinophils express more membrane receptors and release more mediators than blood eosinophils, suggesting that migration from blood to tissue modulates eosinophil phenotype and functions. OBJECTIVE: We postulated that eosinophil passage through endothelial basement membrane, an important step of eosinophil migration into tissue, may be responsible for some of these changes. METHOD: We previously showed that 5-oxo-6, 8, 11, 14-eicosatetraenoic acid (5-oxo-ETE) in combination with IL-5 promotes eosinophil migration through Matrigel, a mouse tumour cell-derived basement membrane. Using this model, we evaluated the effect of trans-Matrigel migration on purified human blood eosinophil expressions of CD44, CD69 and HLA-DR that either increase or appear on activated eosinophils, and releases of peroxidase (EPO), leukotriene (LT) C(4) and granulocyte-monocyte colony stimulating factor (GM-CSF). RESULTS: IL-5, but not 5-oxo-ETE, increased eosinophil expression of CD44 and CD69. Migration of eosinophils through Matrigel significantly increased CD44 expression level over the one induced by IL-5 (P = 0.0001). Migration through Matrigel did not modify CD69 expression compared with the one obtained in the presence of IL-5 alone; however, incubation of eosinophils on Matrigel decreased IL-5-induced CD69 (P = 0.0001). Trans-Matrigel migration did not modify HLA-DR expression, nor EPO, LTC(4) and GM-CSF releases. CONCLUSION: These data show that in vitro trans-Matrigel migration and Matrigel contact modulate eosinophil membrane receptor expression. Consequently, they suggest that migration through basement membrane mediates changes in cell-surface phenotype observed on activated eosinophils and probably prepares them for interactions with tissue components and cells.
Assuntos
Membrana Basal/metabolismo , Movimento Celular/fisiologia , Eosinófilos/metabolismo , Receptores de Hialuronatos/biossíntese , Adolescente , Adulto , Ácidos Araquidônicos/farmacologia , Asma/metabolismo , Asma/fisiopatologia , Membrana Basal/citologia , Membrana Basal/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Movimento Celular/efeitos dos fármacos , Colágeno/farmacologia , Citocinas/efeitos dos fármacos , Citocinas/metabolismo , Combinação de Medicamentos , Eosinófilos/efeitos dos fármacos , Feminino , Volume Expiratório Forçado/fisiologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Humanos , Receptores de Hialuronatos/efeitos dos fármacos , Mediadores da Inflamação/metabolismo , Interleucina-5/farmacologia , Laminina/farmacologia , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Proteoglicanas/farmacologia , Receptores de Superfície Celular/biossíntese , Receptores de Superfície Celular/efeitos dos fármacosRESUMO
BACKGROUND: Pathophysiology of corticosteroid (CS)-resistant asthma remains incompletely understood. OBJECTIVE: To determine if failure of asthma to clinically improve with CS is due to a defective response of airway bronchial inflammation to these drugs. METHODS: Twenty-one asthmatics having a decreased baseline FEV1 that improved >or= 30% with inhaled beta2 agonist got bronchial biopsies before and at the end of an oral CS treatment (methylprednisolone 40 mg daily for 14 days). They were arbitrarily divided into two groups according to baseline FEV1 improvement following this treatment: >or= 23% designated as CS-sensitive (CSS) (n = 10) and < 15% as CS-resistant (CSR) (n = 11). RESULTS: Before oral CS, counts of bronchial mucosa inflammatory cells identified by immunohistochemistry (CD3, MBP, tryptase, CD68, neutrophil elastase and CD25 for lymphocytes, eosinophils, mast cells, macrophages, neutrophils and IL-2 receptors, respectively) were similar in CSS and CSR subjects. Oral CS decreased CD3+ cell counts (medians: 60-20 cells/mm(2); P = 0.014) and MBP+ cell counts (medians: 19-4 cells/mm(2); P = 0.03) in CSS asthmatics, but only tryptase+ cell counts in CSR asthmatics (medians: 30-18 cells/mm(2); P = 0.05). Few bronchial neutrophil elastase+ cells were observed and their counts were similar in the two groups of asthmatics before and when on oral CS (all medians: = 2 cells/mm(2)). CONCLUSIONS: These data show that, in these subjects with moderate to severe asthma, lymphocytes and eosinophils constitute most of the inflammatory cells infiltrating the bronchial mucosa. They also demonstrated that clinical impaired response to CS is associated with a persistent bronchial mucosa cellular infiltrate despite oral CS treatment. Additional studies are required to determine the role of this CS-resistant bronchial inflammation in the impaired asthma clinical response to these drugs.