RESUMO
BACKGROUND: The low rate of nontuberculous mycobacteria (NTM) among Brazilian patients with cystic fibrosis (CF) may be due to cross-reactive Bacille Calmette-Guérin (BCG) vaccination. In the present pilot study, we aimed to compare the lymphocyte responses against Mycobacterium tuberculosis(Mtb) and Mycobacterium bovis (BCG) in BCG-vaccinated CF patients and healthy controls. METHODS: The lymphocyte responses of CF patients (n = 10) and healthy controls (n = 10) were assessed in terms of lymphocyte proliferation index (LPI), using flow cytometry. Median rates of each cell subtype - CD4, CD8, γδ T cells and CD19 (B) cells - were also determined. RESULTS: Median LPIs (CF vs. controls) were 22.9% vs. 13.0% (p = 0.481) and 23.1% vs. 17.6% (p = 0.481), upon stimulation with Mtb and BCG, respectively. Both groups had a predominant CD4 T cell response to Mtb (median rate = 82.5% vs. 79.7%; p = 0.796) and BCG (LPI = 84.3% vs. 83.0%; p = 0.853), which were significantly higher than the CD8, CD19 and γδ responses within both groups. CF patients tended to have a higher CD8 T cell response upon stimulation with the phytohemagglutinin mitogen than healthy controls (median rate = 42.8% vs. 31.7%, p = 0.075). CONCLUSION: The responses of BCG-vaccinated CF patients to Mtb and BCG are at least similar to those of healthy individuals. These are probably memory responses elicited by the BCG vaccination, which can cross-react with NTM and may explain the low frequency of NTM lung infection in our CF center.
Assuntos
Vacina BCG , Fibrose Cística , Imunidade Inata , Ativação Linfocitária/imunologia , Subpopulações de Linfócitos/imunologia , Infecções por Mycobacterium não Tuberculosas , Tuberculose/prevenção & controle , Adolescente , Vacina BCG/imunologia , Vacina BCG/uso terapêutico , Brasil/epidemiologia , Fibrose Cística/diagnóstico , Fibrose Cística/epidemiologia , Fibrose Cística/imunologia , Feminino , Humanos , Imunidade Inata/efeitos dos fármacos , Imunidade Inata/imunologia , Memória Imunológica , Masculino , Infecções por Mycobacterium não Tuberculosas/epidemiologia , Infecções por Mycobacterium não Tuberculosas/prevenção & controle , Mycobacterium bovis/imunologia , Mycobacterium tuberculosis/imunologia , Projetos Piloto , Vacinação/métodosRESUMO
BACKGROUND: The upper airways (UAW) are a niche and a reservoir of Pseudomonas aeruginosa strains that cause chronic infection of the lower airways (LAW) in cystic fibrosis (CF). Here, we assessed the role of anti-P. aeruginosa immunoglobulin A (IgA) and IgG antibodies in upper and lower airway infections in cystic fibrosis patients. METHODS: Nasal lavage fluid and induced sputum samples of 40 CF patients were microbiologically cultured. We searched for correlations between anti-P. aeruginosa IgA and IgG levels, measured by enzyme-linked immunosorbent assay (optical density), and unspecific immune mediators in both specimens. RESULTS: Anti-P. aeruginosa IgA (median optical density: 0.953 vs 0.298) and IgG (0.120 vs 0.059) were significantly higher in nasal lavage than in sputum, but not significantly different between patients with and without chronic P. aeruginosa infection in UAW. Matrix metallopeptidase-9 (MMP-9) in nasal lavage and neutrophil elastase (NE) in sputum were predictors of IgA in nasal lavage and IgA in sputum, respectively. IgA was a predictor of myeloperoxidase (MPO) in nasal lavage. Tissue inhibitor of metalloproteinases-1 (TIMP-1) was a predictor of IgG in sputum. IgG, TIMP-1, and NE in sputum were predictors of IgG in nasal lavage. CONCLUSION: The anti-P. aeruginosa IgA response was more prominent in CF patients' UAW, indicating a lower degree of inflammatory responses. Proteases may play a role in the anti-P. aeruginosa humoral response in the upper and LAW, and anti-P. aeruginosa IgG may be involved in the crosstalk between upper and lower airways in cystic fibrosis patients.
Assuntos
Fibrose Cística/microbiologia , Pseudomonas aeruginosa , Sistema Respiratório/microbiologia , Adulto , Formação de Anticorpos , Fibrose Cística/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Elastase de Leucócito , Masculino , Lavagem Nasal , Peptídeo Hidrolases , Peroxidase , Infecções por Pseudomonas/microbiologia , Escarro/microbiologia , Inibidor Tecidual de Metaloproteinase-1RESUMO
BACKGROUND AND OBJECTIVES: The mechanisms leading to low effectiveness of the humoral immune response against P. aeruginosa in cystic fibrosis (CF) are poorly understood. The aim of the present study was to assess the avidity maturation of specific antipseudomonal IgG before and during the development of chronic lung infection in a cohort of Danish CF patients. METHODS: Avidity maturation was assessed against a pooled P. aeruginosa antigen (St-Ag) and against P. aeruginosa alginate in 10 CF patients who developed chronic lung infection and 10 patients who developed intermittent lung colonization, using an ELISA technique with the thiocyanate elution method. Avidity was quantitatively determined by calculating the avidity Constant (Kav). RESULTS: IgG avidity to St-Ag significantly increased at the onset (Median Kav=2.47) and one year after the onset of chronic infection (Median Kav=3.27), but did not significantly changed in patients who developed intermittent colonization. IgG avidity against alginate did not significantly change over the years neither in patients who developed chronic lung infection (Median Kav=3.84 at the onset of chronic infection), nor in patients who developed intermittent colonization. CONCLUSION: IgG avidity to P. aeruginosa alginate does not significantly enhance as chronic lung infection progresses. This probably plays a role in the difficulty to mount an effective opsonophagocytic killing to clear mucoid P. aeruginosa infection in CF.
Assuntos
Biofilmes , Fibrose Cística , Imunoglobulina G/sangue , Infecções por Pseudomonas , Pseudomonas aeruginosa , Adolescente , Adulto , Criança , Doença Crônica , Fibrose Cística/imunologia , Fibrose Cística/microbiologia , Dinamarca , Feminino , Humanos , Imunidade Humoral , Masculino , Infecções por Pseudomonas/diagnóstico , Infecções por Pseudomonas/imunologia , Infecções por Pseudomonas/fisiopatologia , Pseudomonas aeruginosa/isolamento & purificação , Pseudomonas aeruginosa/fisiologia , Reprodutibilidade dos TestesRESUMO
P. aeruginosa chronic lung infection is the major cause of morbidity and mortality in patients with cystic fibrosis (CF), and is characterized by a biofilm mode of growth, increased levels of specific IgG antibodies and immune complex formation. However, despite being designed to combat this infection, such elevated humoral response is not associated with clinical improvement, pointing to a lack of anti-pseudomonas effectiveness. The mode of action of specific antibodies, as well as their structural features, and even the background involving B-cell production, stimulation and differentiation into antibody-producing cells in the CF airways are poorly understood. Thus, the aim of this review is to discuss studies that have addressed the intrinsic features of the humoral immune response and provide new insights regarding its insufficiency in the CF context.
Assuntos
Biofilmes , Fibrose Cística/imunologia , Imunidade Humoral , Infecções por Pseudomonas/prevenção & controle , Pseudomonas aeruginosa , Infecções Respiratórias/prevenção & controle , Fibrose Cística/microbiologia , Fibrose Cística/terapia , Humanos , Infecções por Pseudomonas/etiologia , Infecções Respiratórias/etiologiaRESUMO
OBJECTIVE: To evaluate the results of the tests used to identify the IgE mediated sensitization to Aspergillus fumigatus in patients with cystic fibrosis. METHODS: This is a cross-sectional descriptive study with a convenience sample of 86 patients diagnosed with cystic fibrosis in the Reference Service in Cystic Fibrosis at a tertiary teaching hospital. The following tests were performed to assess the sensitization to A. fumigatus in patients with cystic fibrosis: Total serum IgE, eosinophil count, fungus detection through oropharyngeal swab or sputum culture, serum-specific IgE, and immediate-type hypersensitivity (IgE) skin tests. We compared the results of the different tests performed. RESULTS: In 33 (38.4%) patients with cystic fibrosis, with ages ranging from 1 to 33 years (median of 8 years), the IgE-mediated A. fumigatus sensitization test results were: in 16 patients, there was an increase in serum-specific IgE (>0.35 kU/L); in 23, skin tests were positive; and six had sensitization in both tests. We observed two patients with eosinophilia (>1,000 eosinophils/mm3) and seven with increasing total serum IgE (>1,000 IU/mL), all of whom obtained negative results in skin tests and had no IgE increase specific to A. fumigatus. A. fumigatus was not detected in oropharyngeal swabs and/or sputum culture of any patients. CONCLUSIONS: We conclude that, among the tests used to assess sensitization to A. fumigatus in cystic fibrosis patients, both serum-specific IgE and immediate-type hypersensitivity (IgE) skin tests are required. Serum eosinophilia and respiratory secretion culture were not essential in this study.
OBJETIVO: Avaliar os resultados dos exames utilizados para identificar a sensibilização IgE-mediada ao Aspergillus fumigatus em pacientes com fibrose cística. MÉTODOS: Estudo transversal descritivo com amostra de conveniência de 86 pacientes com fibrose cística, acompanhados em Serviço de Referência de Fibrose Cística de hospital universitário terciário. Realizaram-se exames para avaliar sensibilização ao A. fumigatus em pacientes com fibrose cística: IgE sérica total, contagem de eosinófilos sanguíneos, identificação do fungo por swab de orofaringe ou por cultura de escarro, IgE sérica específica e testes cutâneos de hipersensibilidade imediata. Foram comparados os resultados dos diferentes exames realizados. RESULTADOS: Em 33 (38,4%) pacientes com fibrose cística, com faixa etária de 1 a 33 anos (mediana de 8 anos), os resultados dos exames sobre sensibilização IgE mediada ao A. fumigatus foram: em 16 pacientes, aumento de IgE sérica específica (>0,35 kU/L); em 23, positividade aos testes cutâneos; e seis mostraram sensibilização a partir dos dois exames. Foram observados dois pacientes com eosinofilia (>1.000 eosinófilos/mm3) e sete com aumento de IgE sérica total (>1.000 UI/mL), sem que esses apresentassem positividade aos testes cutâneos ou aumento de IgE específica ao A. fumigatus. Em nenhum paciente foi isolado A. fumigatus no swab de orofaringe e/ou na cultura de escarro. CONCLUSÕES: Concluímos que, entre os exames para avaliar a sensibilização ao A. fumigatus na fibrose cística, são necessários os teste cutâneos de hipersensibilidade imediata e a dosagem de IgE sérica específica ao A. fumigatus. A eosinofilia sérica e a cultura de secreções respiratórias não foram essenciais neste estudo.
Assuntos
Aspergillus fumigatus/imunologia , Fibrose Cística/imunologia , Hipersensibilidade Imediata/diagnóstico , Imunoglobulina E/imunologia , Adolescente , Criança , Pré-Escolar , Estudos Transversais , Feminino , Humanos , Lactente , MasculinoRESUMO
RESUMO Objetivo: Avaliar os resultados dos exames utilizados para identificar a sensibilização IgE-mediada ao Aspergillus fumigatus em pacientes com fibrose cística. Métodos: Estudo transversal descritivo com amostra de conveniência de 86 pacientes com fibrose cística, acompanhados em Serviço de Referência de Fibrose Cística de hospital universitário terciário. Realizaram-se exames para avaliar sensibilização ao A. fumigatus em pacientes com fibrose cística: IgE sérica total, contagem de eosinófilos sanguíneos, identificação do fungo por swab de orofaringe ou por cultura de escarro, IgE sérica específica e testes cutâneos de hipersensibilidade imediata. Foram comparados os resultados dos diferentes exames realizados. Resultados: Em 33 (38,4%) pacientes com fibrose cística, com faixa etária de 1 a 33 anos (mediana de 8 anos), os resultados dos exames sobre sensibilização IgE mediada ao A. fumigatus foram: em 16 pacientes, aumento de IgE sérica específica (>0,35 kU/L); em 23, positividade aos testes cutâneos; e seis mostraram sensibilização a partir dos dois exames. Foram observados dois pacientes com eosinofilia (>1.000 eosinófilos/mm3) e sete com aumento de IgE sérica total (>1.000 UI/mL), sem que esses apresentassem positividade aos testes cutâneos ou aumento de IgE específica ao A. fumigatus. Em nenhum paciente foi isolado A. fumigatus no swab de orofaringe e/ou na cultura de escarro. Conclusões: Concluímos que, entre os exames para avaliar a sensibilização ao A. fumigatus na fibrose cística, são necessários os teste cutâneos de hipersensibilidade imediata e a dosagem de IgE sérica específica ao A. fumigatus. A eosinofilia sérica e a cultura de secreções respiratórias não foram essenciais neste estudo.
ABSTRACT Objective: To evaluate the results of the tests used to identify the IgE mediated sensitization to Aspergillus fumigatus in patients with cystic fibrosis. Methods: This is a cross-sectional descriptive study with a convenience sample of 86 patients diagnosed with cystic fibrosis in the Reference Service in Cystic Fibrosis at a tertiary teaching hospital. The following tests were performed to assess the sensitization to A. fumigatus in patients with cystic fibrosis: Total serum IgE, eosinophil count, fungus detection through oropharyngeal swab or sputum culture, serum-specific IgE, and immediate-type hypersensitivity (IgE) skin tests. We compared the results of the different tests performed. Results: In 33 (38.4%) patients with cystic fibrosis, with ages ranging from 1 to 33 years (median of 8 years), the IgE-mediated A. fumigatus sensitization test results were: in 16 patients, there was an increase in serum-specific IgE (>0.35 kU/L); in 23, skin tests were positive; and six had sensitization in both tests. We observed two patients with eosinophilia (>1,000 eosinophils/mm3) and seven with increasing total serum IgE (>1,000 IU/mL), all of whom obtained negative results in skin tests and had no IgE increase specific to A. fumigatus. A. fumigatus was not detected in oropharyngeal swabs and/or sputum culture of any patients. Conclusions: We conclude that, among the tests used to assess sensitization to A. fumigatus in cystic fibrosis patients, both serum-specific IgE and immediate-type hypersensitivity (IgE) skin tests are required. Serum eosinophilia and respiratory secretion culture were not essential in this study.
Assuntos
Humanos , Masculino , Feminino , Lactente , Pré-Escolar , Criança , Adolescente , Aspergillus fumigatus/imunologia , Imunoglobulina E , Fibrose Cística/imunologia , Hipersensibilidade Imediata/diagnóstico , Estudos TransversaisRESUMO
We assessed the diagnostic ability of an enzyme-linked immunosorbent assay test for measurement of specific secretory IgA (sIgA) in saliva to identify cystic fibrosis (CF) patients with Pseudomonas aeruginosa chronic lung infection and intermittent lung colonization. A total of 102 Brazilian CF patients and 53 healthy controls were included. Specific serum IgG response was used as a surrogate to distinguish CF patients according to their P. aeruginosa colonization/infection status. The rate of sIgA positivity was 87.1% in CF chronically infected patients (median value = 181.5 U/mL), 48.7% in intermittently colonized patients (median value = 45.8 U/mL) and 21.8% in free of infection patients (median value = 22.1 U/mL). sIgA levels in saliva were significantly associated with serum P. aeruginosa IgG and microbiological culture results. The sensitivity, specificity, PPV and NPV for differentiation between presence and absence of chronic lung infection were 87%, 63%, 51% and 92%, respectively. Measurement of sIgA in saliva may be used for screening patients in risk of developing P. aeruginosa chronic lung infection in CF and possibly also for paranasal sinusitis, and, most importantly, to efficiently rule out chronic P. aeruginosa lung infection.
Assuntos
Anticorpos Antibacterianos/análise , Fibrose Cística/diagnóstico , Imunoglobulina A Secretora/análise , Imunoglobulina G/análise , Infecções Oportunistas/diagnóstico , Infecções por Pseudomonas/diagnóstico , Saliva/química , Adolescente , Adulto , Estudos de Casos e Controles , Criança , Pré-Escolar , Estudos Transversais , Fibrose Cística/complicações , Fibrose Cística/imunologia , Fibrose Cística/microbiologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Pulmão/imunologia , Pulmão/microbiologia , Pulmão/patologia , Masculino , Infecções Oportunistas/complicações , Infecções Oportunistas/imunologia , Infecções Oportunistas/microbiologia , Infecções por Pseudomonas/complicações , Infecções por Pseudomonas/imunologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa , Saliva/imunologia , Saliva/microbiologia , Sensibilidade e EspecificidadeRESUMO
Cystic fibrosis (CF) is an autosomal recessive disorder caused by mutations in the gene that codes for the CF trans-membrane conductance regulator. These mutations result in abnormal secretions viscous airways of the lungs, favoring pulmonary infection and inflammation in the middle of neutrophil recruitment. Recently it was described that neutrophils can contribute with disease pathology by extruding large amounts of nuclear material through a mechanism of cell death known as Neutrophil Extracellular Traps (NETs) into the airways of patients with CF. Additionally, NETs production can contribute to airway colonization with bacteria, since they are the microorganisms most frequently found in these patients. In this review, we will discuss the implication of individual or mixed bacterial infections that most often colonize the lung of patients with CF, and the NETs role on the disease.
Assuntos
Infecções Bacterianas/patologia , Fibrose Cística/complicações , Fibrose Cística/imunologia , Armadilhas Extracelulares , Infiltração de Neutrófilos , Bactérias/imunologia , Infecções Bacterianas/microbiologia , Coinfecção/microbiologia , Coinfecção/patologia , Fibrose Cística/patologia , HumanosRESUMO
AIMS: Recurrent infections and activation of the inflammatory response affect the prognosis of cystic fibrosis (CF). We investigated the relationship between inflammatory response, infection, and pulmonary function in CF. MAIN METHODS: A clinical-cross-sectional study was conducted with 86 subjects: control group (CG, n=31, the same age and sex of the CF group), and CF group (CFG, n=55, age: 1-16 years), further distributed into CFG negative or positive bacteriology (CFGB(-)/CFGB(+)), and CFG negative or positive Pseudomonas aeruginosa (CFGPa(-)/CFGPa(+)). Using the Wald test, multiple linear regression (95% confidence interval) was performed between CG and CFG, and between CG and each of the CF subgroups (CFGB(-)/CFGB(+) and CFGPa(-)/CFGPa(+)). The inflammatory markers evaluated were myeloperoxidase (MPO), adenosine deaminase (ADA) activities, interleukin-1beta (IL-1ß), tumor necrosis factor-alpha (TNF-α), C-reactive protein (CRP), nitric oxide metabolites (NOx) levels, and total and differential leukocyte counts. KEY FINDINGS: After adjusting for sex and age, CFG compared to CG revealed an increase of MPO, IL-1ß (P<0.001 in all subgroups), and CRP: CFG (P=0.002), CFGB(-) (P=0.007), CFGB(+) (P=0.009), CFGPa(-) (P=0.004) and CFGPa(+) (P=0.020). NOx (P=0.001, P<0.001), leukocytes (P=0.002, P=0.001), and neutrophils (P=0.003, P<0.001) were increased in CFGB(+) and CFGPa(+), respectively. A negative correlation between FEV1 and leukocytes (P=0.008) and FEV1 and neutrophils (P=0.031) resulted in CFG. SIGNIFICANCE: The inflammatory response characterized by the increase of MPO, IL-1ß, and CRP is determinant for CF. Also leukocytosis due to neutrophilia determines the pulmonary function deficiency in this disease.
Assuntos
Fibrose Cística/complicações , Pneumonia/complicações , Infecções por Pseudomonas/complicações , Pseudomonas aeruginosa/imunologia , Adolescente , Criança , Pré-Escolar , Estudos Transversais , Fibrose Cística/diagnóstico , Fibrose Cística/imunologia , Fibrose Cística/microbiologia , Feminino , Humanos , Lactente , Pulmão/imunologia , Pulmão/microbiologia , Masculino , Pneumonia/diagnóstico , Pneumonia/imunologia , Pneumonia/microbiologia , Infecções por Pseudomonas/diagnóstico , Infecções por Pseudomonas/imunologia , Pseudomonas aeruginosa/isolamento & purificaçãoRESUMO
BACKGROUND: A systematic literature review of the last 40 years on the research of serum antibodies to Pseudomonas aeruginosa in cystic fibrosis and its utility as a diagnostic tool. METHODS: Research papers in English, Portuguese, and Spanish were accessed through electronic databases (PubMed, Medline, LILACS, and SciELO). RESULTS: 26 studies were assessed. ELISA technique was the most commonly used technique to detect serum P. aeruginosa antibodies. The most consistent results were those in which the response against the antigen St-Ag:1-17 was evaluated. The accuracy levels of the ELISA technique remain controversial, but most studies showed a good correlation between antibody titers and microbiological culture. CONCLUSIONS: The detection of serum antibodies to P. aeruginosa shows capacity for early detection of this pathogen and potential utility and viability of incorporation in the diagnostic routine of patients with cystic fibrosis.