RESUMO
SETTING: Information about the sputum cells of pulmonary tuberculosis (PTB) patients is scarce. The analysis of sputum cells using optical microscopy (OM) is a well-established method, but it has some serious limitations. OBJECTIVE: To establish a new flow cytometry (FC) protocol for the leucocyte evaluation of sputum samples from PTB patients. DESIGN: A new FC protocol using 0.1% dithiothreitol and 0.5% paraformaldehyde was developed to fluidise sputum samples and kill Mycobacterium tuberculosis, respectively, to allow the analysis of sputum samples collected from TB patients. The protocol was validated by comparing it with OM, and the cellularity of 30 sputum samples from patients with PTB was evaluated. RESULTS: The comparison between leucocyte subsets analysed using OM and FC showed agreement. Immunophenotyping of leucocytes from sputum samples showed that neutrophils (95.7%) comprised the largest proportion of sputum cells, followed by monocytes/macrophages (2.6%) and lymphocytes (1.6%). Among the total T-lymphocytes (100%), 12.3% were T-helper cells, 24.1% were cytotoxic T-cells and 62.9% were gamma/delta T; none of the T lymphocytes had the CD4+/CD8+ phenotype. CONCLUSION: FC is a useful method for evaluating the different subtypes of leucocytes present in the sputum samples of PTB patients.
Assuntos
Leucócitos/imunologia , Escarro/citologia , Escarro/imunologia , Tuberculose Pulmonar/imunologia , Adolescente , Adulto , Brasil , Estudos de Casos e Controles , Feminino , Citometria de Fluxo , Humanos , Masculino , Microscopia , Pessoa de Meia-Idade , Mycobacterium tuberculosis/imunologia , Adulto JovemRESUMO
BACKGROUND: Immune response has been implicated in the pathogenesis of chronic obstructive pulmonary disease (COPD) and asthma. Pentraxin 3 (PTX3) is a multifunctional pattern recognition protein and an important component of the innate immune system that can be assessed in blood and induced sputum. OBJECTIVE: To determine whether PTX3 measured in induced sputum could discriminate patients with COPD from patients with asthma. METHODS: A cross-sectional study of 68 participants (27 with COPD, 25 with asthma, and 16 healthy controls) was performed. At study inclusion sputum was collected and total and differential cell numbers and PTX3 levels were determined. RESULTS: Pentraxin 3 was detected in 89% of patients with COPD, 56% of patients with asthma, and 19% of controls (P = .001). It discriminated participants with COPD (24.6 ng/mL, 0-384 ng/mL) from controls (0 ng/mL, 0-36 ng/mL, P < .001) and from participants with asthma (1.2 ng/mL, 0-100 ng/mL, P = .01; area under the receiver operating curve 0.82 [0.71-0.94]). Regression analyses determined that sputum PTX3 and neutrophil counts were independently associated with COPD. In addition, PTX3 levels were independently associated with COPD severity. CONCLUSION: Pentraxin 3 sputum levels are increased in patients with COPD and has good power to discriminate these patients from patients with asthma and healthy individuals.
Assuntos
Asma/imunologia , Proteína C-Reativa/imunologia , Doença Pulmonar Obstrutiva Crônica/imunologia , Componente Amiloide P Sérico/imunologia , Escarro/imunologia , Adolescente , Adulto , Idoso , Asma/fisiopatologia , Contagem de Células , Estudos Transversais , Feminino , Volume Expiratório Forçado , Humanos , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Índice de Gravidade de Doença , Escarro/citologia , Adulto JovemRESUMO
Sputum and sera from 134 patients screened for tuberculosis (TB) were analyzed to investigate TB and paracoccidioidomycosis (PCM). Of these patients, 11 (8.2%) were confirmed to have TB, but six (4.5%) were positive only for PCM. All patients with PCM presented anti-43-kDa-component antibodies in Western blotting (WB) assays, while in the TB-positive patients these antibodies did not appear. This preliminary study suggests WB as a potential tool for differential laboratory diagnosis between TB and PCM.
Assuntos
Anticorpos Antifúngicos/análise , Western Blotting/métodos , Técnicas de Laboratório Clínico/métodos , Testes Diagnósticos de Rotina/métodos , Paracoccidioidomicose/diagnóstico , Tuberculose Pulmonar/diagnóstico , Adulto , Idoso , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Soro/imunologia , Escarro/imunologiaRESUMO
BACKGROUND: Recently, Achromobacter xylosoxidans has been related to chronic lung diseases in patients suffering from cystic fibrosis (CF), but its involvement has not been elucidated. Some virulence properties of A. xylosoxidans isolated from Brazilian patients with CF were revealed in this work. METHODS: This study examined the production of a cytotoxic factor of A. xylosoxidans capable of stimulating the secretion of inflammatory cytokines (IL-6 and IL-8) from lung mucoepidermoid carcinoma cells (NCI-H292). The cytokines were measured using enzyme-linked immunosorbent (ELISA) assays. To investigate whether the cytotoxic factors may be endotoxins, they were treated with polymyxin B. RESULTS: The culture supernatants of all A. xylosoxidans produced a heat stable, active cytotoxin in NCI-H292 cells capable of leading to intracellular vacuoles and subsequent cell contact loss, chromatin condensation, a picnotic nucleus and cell death. There was a higher concentration of proinflammatory cytokines in the NCI-H292 cells after 24 h of incubation, with the fraction greater than 50 kDa from the culture supernatant. The cytotoxin activity remained even after treatment with polymyxin B, which suggested that the release of IL-6 and IL-8 was not stimulated by lipopolysaccharide (LPS). CONCLUSION: The cytotoxic factor produced by A. xylosoxidans may represent an important virulence factor, which when associated with CF chronic lung inflammation, may cause tissue damage and decline of lung function.
Assuntos
Achromobacter denitrificans/metabolismo , Fibrose Cística/microbiologia , Endotoxinas/metabolismo , Infecções por Bactérias Gram-Negativas/imunologia , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Achromobacter denitrificans/imunologia , Achromobacter denitrificans/patogenicidade , Brasil , Carcinoma Mucoepidermoide , Linhagem Celular Tumoral , Sobrevivência Celular/imunologia , Meios de Cultivo Condicionados/toxicidade , Fibrose Cística/imunologia , Endotoxinas/imunologia , Infecções por Bactérias Gram-Negativas/metabolismo , Humanos , Técnicas In Vitro , Interleucina-6/imunologia , Interleucina-8/imunologia , Pneumonia/imunologia , Pneumonia/metabolismo , Pneumonia/microbiologia , Escarro/imunologia , Escarro/microbiologia , VirulênciaRESUMO
Interleukin (IL) 10 and interferon-gamma (IFN-) levels in induced sputum supernatants of 21 tuberculosis (TB) patients at diagnosis and during chemotherapy were correlated to recurrence rates. IL-10 decreased until day 60 of treatment (T60), and between T60 and T180 it increased again in 7 cases (Pattern 1) and further decreased in 14 cases (Pattern 2). Follow-up of 69 months was performed in 20/21 cases; 6 had recurrence of TB, of which 5/7 (71%) had Pattern 1 and 1/13 (7.7%) Pattern 2 (OR 30.0, 95%CI 2.19411.3, P 0.0072). This was not observed for IFN-. High IL-10 levels at the end of treatment may function as a risk factor for TB recurrence.
Assuntos
Antituberculosos/uso terapêutico , Interferon gama/imunologia , Interleucina-10/imunologia , Tuberculose/imunologia , Adulto , Feminino , Seguimentos , Humanos , Masculino , Recidiva , Fatores de Risco , Escarro/imunologia , Tuberculose/tratamento farmacológico , Adulto JovemRESUMO
BACKGROUND: Currently, there are no studies of well-characterized severe asthmatics in Brazil. We aimed to study a population of severe treated asthmatics still uncontrolled to characterize them and define possible phenotypes. METHODS: Descriptive cross-sectional outpatient study of severe asthmatics, evaluating functional and inflammatory markers, health-related quality of life, anxiety and depression symptoms, clinical control status, and characteristics related to atopy, age of asthma onset, induced sputum eosinophil levels, and airflow limitation. We also grouped the subgroups characteristics to identify phenotypes. The study is registered on ClinicalTrial.gov NCT 01089322. RESULTS: From 128 eligible patients with severe/uncontrolled asthma, 74 fulfilled the inclusion criteria. The cohort was comprised of 85% women, frequently with a body mass index higher than 31 kg m(-2), atopy (60%), early-onset disease (50%), sputum eosinophilia (80%), comorbidities, and reduced quality of life. Nonatopics had significant higher asthma onset (19 y.a.) and twice level of induced sputum eosinophil. Late-onset patients had significantly less atopy (57%) and higher levels of induced sputum eosinophils. Non-eosinophilics had lower levels of inflammatory markers. Patients with airflow limitation had more intensive care unit admissions (56%) and 1.5 times more airway resistance. Subgroups characteristics identified a priori four well-characterized phenotypes, with 55% presenting sputum eosinophilia. CONCLUSION: Our data emphasize the high burden of disease, the persistence of inflammation and the existence of clinical possible phenotypes population sharing common features with published cohorts. Despite the necessity of further investigation into pathogenic mechanisms, this study with clinically difficult patient group may help to improve future asthma care.
Assuntos
Resistência das Vias Respiratórias/imunologia , Asma/imunologia , Eosinófilos/imunologia , Hipersensibilidade Imediata/imunologia , Escarro/imunologia , Adolescente , Adulto , Idoso , Ansiedade/epidemiologia , Ansiedade/fisiopatologia , Asma/epidemiologia , Asma/genética , Asma/fisiopatologia , Índice de Massa Corporal , Brasil/epidemiologia , Ensaios Clínicos como Assunto , Estudos de Coortes , Estudos Transversais , Depressão/epidemiologia , Depressão/fisiopatologia , Feminino , Humanos , Hipersensibilidade Imediata/epidemiologia , Hipersensibilidade Imediata/genética , Hipersensibilidade Imediata/fisiopatologia , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/metabolismo , Fenótipo , Testes de Função Respiratória , Índice de Gravidade de Doença , Distribuição por Sexo , Adulto JovemRESUMO
The identification of appropriate laboratory measures to confirm clinical hypotheses is important in routine paracoccidioidomycosis medical care. The clinical records and laboratory reports of 401 paracoccidioidomycosis patients attended at the Tropical Diseases Area, Faculdade de Medicina de Botucatu, from 1974 to 2008 were reviewed. Direct mycological (DM), cell block (CB), histopathological (HP), and double immunodiffusion (DID) tests were evaluated before treatment. Typical Paracoccidioides brasiliensis yeast forms were observed in clinical specimens of 86% of the patients, but 14% were detected only by serological test. DM of 51 different tissue specimens produced 74.5% sensitivity, and 62.5% sensitivity was observed in 112 sputum samples. CB in 483 sputum samples generated 55.3% sensitivity. HP performed in 239 samples from different tissues revealed 96.7% sensitivity. Serology carried out in 351 patients and 200 healthy controls provided 90.0% sensitivity, 100.0% specificity, 100.0% positive predictive value, 85.1% negative predictive value and 93.6% accuracy. Comparisons of laboratory measurements performed in the same patient showed that sensitivity decreases from HP to DID to CB and DM, with the last two assays providing similar sensitivities. This study demonstrated that P. brasiliensis identification by HP, CB, and/or DM associated with DID is sufficient to establish the laboratorial diagnosis of paracoccidioidomycosis in practically all cases.
Assuntos
Imunodifusão , Paracoccidioides/isolamento & purificação , Paracoccidioidomicose/imunologia , Escarro/imunologia , Adulto , Brasil/epidemiologia , Testes Diagnósticos de Rotina , Feminino , Hospitais Universitários , Humanos , Imunodifusão/métodos , Masculino , Pessoa de Meia-Idade , Paracoccidioidomicose/diagnóstico , Paracoccidioidomicose/epidemiologia , Estudos RetrospectivosRESUMO
Se realizó un estudio descriptivo y transversal para caracterizar a los 155 pacientes con tuberculosis en la provincia de Santiago de Cuba durante los años 2005-2007. Hubo un ligero incremento de la tasa de incidencia de la enfermedad en ese trienio, con predominio en los municipios de Santiago de Cuba, Palma Soriano y Julio Antonio Mella (en este último aumentó de forma significativa en el 2007). Se observó, de forma general, que los afectados tenían entre 2 y 3 factores de riesgo y la mayoría pertenecieron a los grupos etarios de más de 55 años. Primaron la tuberculosis pulmonar y la positividad del bacilo ácido-alcohol resistente. En la atención primaria se diagnosticó el mayor número de personas infectadas, las cuales tenían menos de 23 días de haber estado experimentando los síntomas. Hubo 5 pacientes que presentaron coinfección con el virus de inmunodeficiencia adquirida.
A descriptive and cross-sectional study was carried out to characterize the 155 patients with tuberculosis in Santiago de Cuba province during the years 2005-2007. There was a slight increase of the incidence rate of the disease in that triennium, with prevalence in Santiago de Cuba, Palma Soriano and Julio Antonio Mella municipalities (in this last one it increased in a significant way in 2007). In general it was observed, that the affected ones had between 2 and 3 risk factors and most of them were in the age group of more than 55 years. The lung tuberculosis and the positivity of the acid-alcohol resistant bacillus prevailed. The highest number of infected people who had less than 23 days of experiencing the symptoms, was diagnosed in the primary care. There were 5 patients that presented co-infection with the acquired immunodeficiency virus.
Assuntos
Humanos , Masculino , Feminino , Escarro/citologia , Escarro/imunologia , HIV , Programas Nacionais de Saúde , Atenção Primária à Saúde , Testes Respiratórios/métodos , Fatores de Risco , Tuberculose , Tuberculose Pulmonar , Estudos Transversais , Epidemiologia Descritiva , Estudos Observacionais como AssuntoRESUMO
BACKGROUND: It is noteworthy that there is a clear clinical, epidemiological and pathophysiological association between upper and lower airway inflammation in rhinitis and asthma. OBJECTIVE: The aim of this study was to compare the eosinophil counts in induced sputum and nasal lavage fluids in asthma, checking their association and the accuracy of nasal eosinophilia as a predictor of sputum eosinophilia by a cross-sectional study. METHODS: The clinical evaluation, asthma control questionnaire (ACQ), pre- and post-bronchodilator spirometry, nasal and sputum sample was performed. The nasal eosinophilia was analysed by a receiver operating curve and logistic regression model. RESULTS: In 140 adults, the post-bronchodilator forced expiratory volume in 1 s (FEV(1)) did not differ between patients with or without sputum eosinophilia (0.18). After adjusted for upper airway symptoms, age, ACQ score and post-bronchodilator FEV(1), sputum eosinophilia was associated with 52 times increase in odds of nasal eosinophilia, whereas each 1% increase in bronchodilator response was associated with 7% increase in odds of nasal eosinophilia. CONCLUSION: This study brings further evidence that upper airway diseases are an important component of the asthma syndrome. Furthermore, monitoring of nasal eosinophilia by quantitative cytology may be useful as a surrogate of sputum cytology in as a component of composite measurement for determining airway inflammation.
Assuntos
Asma , Eosinofilia/diagnóstico , Inflamação , Nariz/imunologia , Administração Intranasal , Adulto , Asma/diagnóstico , Asma/imunologia , Asma/fisiopatologia , Estudos Transversais , Eosinofilia/imunologia , Eosinófilos/citologia , Eosinófilos/imunologia , Feminino , Humanos , Inflamação/imunologia , Inflamação/fisiopatologia , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Líquido da Lavagem Nasal , Escarro/imunologiaRESUMO
INTRODUCTION: Common variable immunodeficiency is characterized by defective antibody production and recurrent pulmonary infections. Intravenous immunoglobulin is the treatment of choice, but the effects of Intravenous immunoglobulin on pulmonary defense mechanisms are poorly understood. OBJECTIVE: The aim of this study was to verify the impact of intravenous immunoglobulin on the physical properties of the sputum and on inflammatory alterations in the airways of patients with Common variable immunodeficiency associated with bronchiectasis. METHOD: The present study analyzed sputum physical properties, exhaled NO, inflammatory cells in the sputum, and IG titers in 7 patients with Common variable immunodeficiency and bronchiectasis with secretion, immediately before and 15 days after Intravenous immunoglobulin. A group of 6 patients with Common variable immunodeficiency and bronchiectasis but no sputum was also studied for comparison of the basal IgG level and blood count. The 13 patients were young (age=36+/-17 years) and comprised predominantly of females (n=11). RESULTS: Patients with secretion presented significantly decreased IgG and IgM levels. Intravenous immunoglobulin was associated with a significant decrease in exhaled NO (54.7 vs. 40.1 ppb, p<0.05), sputum inflammatory cell counts (28.7 vs. 14.6 cells/mm(3), p<0.05), and a significant increase in respiratory mucus transportability by cough (42.5 vs. 65.0 mm, p < 0.05). CONCLUSION: We concluded that immunoglobulin administration in Common variable immunodeficiency patients results in significant improvement in indexes of inflammation of the airways with improvement in the transportability of the respiratory mucus by cough.