RESUMO
CONTEXT: Allergic bronchopulmonary mycoses (ABPM) can be due to molds other than Aspergillus fumigatus in patients with cystic fibrosis (pwCF). We aimed to develop immunoassays for the detection of specific IgE (sIgE) directed against five fungal species involved in ABPM: Aspergillus terreus, Scedosporium apiospermum, Lomentospora prolificans, Rasamsonia argillacea, and Exophiala dermatitidis. MATERIALS AND METHODS: Serum samples (n = 356) from 238 pwCF, collected in eight CF care centers in France, Germany, and Italy, were analyzed by dissociated enhanced lanthanide fluorescent immunoassay (DELFIA®) to assess levels of sIgE directed against antigenic extracts of each fungus. Clinical, biological, and radiological data were collected for each episode. One hundred serum samples from healthy blood donors were used as controls. Sera were classified into four groups depending on the level of sIgE according to the quartile repartition calculated for the pwCF population. A score of 4 for values above the 3rd quartile corresponds to an elevated level of sIgE. RESULTS: PwCF showed higher levels of sIgE than controls. Based on criteria from the ABPA-ISHAM working group, with an additional criterion of "a sIgE score of 4 for at least one non-A. fumigatus mold", we were able to diagnose six cases of ABPM. CONCLUSIONS: Using 417 IU/mL as the threshold for total IgE and the same additional criterion, we identified seven additional pwCF with "putative ABPM". Detection of sIgE by DELFIA® showed good analytical performance and supports the role played by non-A. fumigatus molds in ABPM. However, commercially available kits usable in routine practice are needed to improve the diagnosis of ABPM.
Assuntos
Anticorpos Antifúngicos , Fibrose Cística , Fungos , Imunoglobulina E , Humanos , Fibrose Cística/complicações , Imunoglobulina E/sangue , Feminino , Masculino , Adulto , Adulto Jovem , Adolescente , Fungos/imunologia , Fungos/classificação , Fungos/isolamento & purificação , Imunoensaio/métodos , Criança , Anticorpos Antifúngicos/sangue , Itália , França , Alemanha , Pré-Escolar , Pessoa de Meia-Idade , Aspergilose Broncopulmonar Alérgica/diagnóstico , Aspergilose Broncopulmonar Alérgica/imunologia , Aspergilose Broncopulmonar Alérgica/sangueRESUMO
AIMS: Alveolar echinococcosis (AE) is characterized by a chronically progressing hepatic injury caused by Echinococcus multilocularis. Surgery presently remains the best curative option. Currently, biological predictive features derived from the resected specimens are not suitable to assess surgery efficacy. The present study was designed to investigate whether a selection of markers measured on the resected specimens exhibits predictive features related to parasite viability, or to a total elimination of the parasite, in addition to serological markers. METHODS AND RESULTS: In a collaboration between two centres, one in France (Besançon), and one in Switzerland (Bern), samples from 40 AE patients were analysed by microarray and serology techniques, individually. Paired serum samples before and after surgery were obtained for 26 patients. In the sera, a significant decrease in PD-L1 levels was observed after surgery, in addition to anti-Em18 levels. In the liver tissue, low levels of Cluster of Differentiation (CD)-3 were correlated with the absence of serum anti-Em18 after surgery. CONCLUSION: This study showed PD-L1 is promising as a potential serological marker and further confirmed the performance of anti-Em18 serology. Further studies on a larger cohort are needed to confirm the utility of performing systematically microarray on resected liver tissue.
Assuntos
Equinococose Hepática , Equinococose , Antígenos de Helmintos , Equinococose/diagnóstico , Equinococose/cirurgia , Equinococose Hepática/cirurgia , Seguimentos , HumanosRESUMO
AIMS: Following treatment, cystic echinococcosis (CE) exhibits a relatively high relapse rate. Here, we evaluated the value of soluble programmed death-1 (sPD-1), sPD-1 ligand (sPD-L1) and anti-recP29 antibody concentrations, as predictors of early surgical treatment outcomes in young CE-affected patients. METHODS AND RESULTS: This prospective study included 59 Tunisian children (177 plasmas), where CE was surgically treated and monitored for 3 post-operative years. Based on CE post-surgical development, patients were clustered into a 'No relapsed' CE (NRCE; n = 39) and a 'Relapsed' CE (RCE; n = 20) group. Plasma levels of sPD-1, sPD-L1 and anti-recP29 IgG were measured using ELISA. In the NRCE group, sPD-1, sPD-L1 and anti-recP29 IgG concentrations were significantly lower at D365 than at D30. By contrast, in the RCE group, no significant difference was observed between D0, D30 and D365. When considering individual variations, the probability to be 'relapse-free' was 67% and 73% when anti-recP29 IgG and sPD-L1 level, respectively, decreased between D30 and D365. The probability to be 'relapse-free' was 86% when the sPD-1 level decreased between D30 and D365 (P = .003; chi-square test). CONCLUSION: sPD-1 may be a useful biomaker for the early evaluation of surgical procedure efficacy in paediatric CE cases.
Assuntos
Antígeno B7-H1/imunologia , Equinococose/cirurgia , Adolescente , Biomarcadores , Criança , Pré-Escolar , Equinococose/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Estudos Prospectivos , Prevenção Secundária , Resultado do TratamentoRESUMO
Dairy farming is a risk factor for chronic obstructive pulmonary disease (COPD). The aim was to determine predictive markers either in blood samples or in dwelling dust samples by comparing COPD and healthy controls with or without farming activity. Dust was collected and analyzed by real-time quantitative PCR. ELISA and DELFIA® were performed to assay the level of specific IgG and IgE of 10 targeted microorganisms. The dwelling exposure of farmers was higher than in the non-farmers (Especially Eurotium amstelodami and Lichtheimia corymbifera). The IgG response against Wallemia sebi and Saccharopolyspora rectivirgula was more often higher in the farmers than the non-farmers. However, exposure and sensitization to the microorganisms tested cannot explain the occurrence of COPD in the dairy farmers' population. COPD development is probably caused by multiple factors associated with exposure over a period of several years.
Assuntos
Indústria de Laticínios , Exposição Ocupacional/análise , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Adulto , Idoso , Animais , Bactérias/imunologia , Bactérias/isolamento & purificação , Poeira/análise , Fazendeiros , Feminino , França/epidemiologia , Fungos/imunologia , Fungos/isolamento & purificação , Habitação , Humanos , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/sangue , Fatores de RiscoRESUMO
PURPOSE: The aim of this study was to understand the differential acute effects of two distinct wheat-related dusts, such as field or stored wheat dust handling, on workers' health and how those effects evolved at 6 month intervals. METHODS: Exposure, work-related symptoms, changes in lung function, and blood samples of 81 workers handling wheat and 61 controls were collected during the high exposure season and 6 months after. Specific IgG, IgE, and precipitins against 12 fungi isolated from wheat dust were titrated by enzyme-linked immunosorbent assay, dissociation-enhanced lanthanide fluorescence immunoassay, and electrosyneresis. The level of fungi was determined in the workers' environment. Levels of exhaled fraction of nitrogen monoxide (FENO) and total IgE were obtained. Exposure response associations were investigated by mixed logistic and linear regression models. RESULTS: The recent exposure to field wheat dust was associated with a higher prevalence for five of six self-reported airway symptoms and with a lower FENO than those in the control population. Exposure to stored wheat dust was only associated with cough. No acute impact of exposure on respiratory function was observed. Exposure to field wheat dust led to workers' sensitization against the three field fungi Aureobasidum, Cryptococcus, and Phoma, although exposure to storage wheat dust was associated with tolerance. The level of Ig remained stable 6 months after exposure. CONCLUSION: The clinical picture of workers exposed to field or storage wheat dust differed. The systematic characterization of the aerosol microbial profile may help to understand the reasons for those differences.
Assuntos
Doenças dos Trabalhadores Agrícolas/fisiopatologia , Poluentes Ocupacionais do Ar/efeitos adversos , Exposição Ocupacional/efeitos adversos , Doenças Respiratórias/etiologia , Doenças Respiratórias/fisiopatologia , Triticum/efeitos adversos , Adulto , Aerossóis/efeitos adversos , Doenças dos Trabalhadores Agrícolas/etiologia , Antígenos de Fungos/sangue , Poeira/análise , Grão Comestível , Ensaio de Imunoadsorção Enzimática , Feminino , Fungos , Humanos , Entrevistas como Assunto , Modelos Logísticos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/análise , Exposição Ocupacional/análise , Testes de Função Respiratória , SuíçaRESUMO
This research communication aimed to evaluate the level of immunoglobulin E from lactic acid bacteria (LAB) that are used in dairy industries. Previous studies have demonstrated that workers report symptoms of irritation and are frequently IgG-sensitised to LAB. Workers (n = 44) from a probiotic production unity and the control lab were seen by a medical practitioner and responded to an occupational questionnaire. Specific IgE by the DELFIA® technique against 6 strains of LAB were measured on 44 exposed workers and 31 controls sera. Levels of specific IgE were low and no difference was observed between the two groups. This lack of IgE response could be explained by a healthy worker effect, an efficient implementation of personal protective equipment or by an absence of allergic mechanisms to account for the self-reported irritative symptoms. Despite the high concentrations of LAB, preventive measures are effective enough to guarantee no allergic effect and to prevent other adverse health effects. The implementation of preventive measures to avoid or reduce exposure to dust of LAB, and more generally to milk powder, is recommended in all dairy industry.
Assuntos
Laticínios , Indústria Alimentícia , Imunização , Imunoglobulina E/sangue , Lactobacillales/imunologia , Exposição Ocupacional , Alérgenos/imunologia , Humanos , Imunoglobulina E/imunologia , Exposição Ocupacional/prevenção & controleRESUMO
BACKGROUND: Argan is now used worldwide in numerous cosmetic products. Nine workers from a cosmetic factory were examined in our occupational medicine department, following the diagnosis of a case of hypersensitivity pneumonitis (HP) related to handling of argan cakes. METHODS: Operators were exposed to three forms of argan (crude granulates, powder or liquid) depending on the step of the process. All workers systematically completed standardized questionnaires on occupational and medical history, followed by medical investigations, comprising, in particular, physical examination and chest X-rays, total IgE and a systematic screening for specific serum antibodies directed against the usual microbial agents of domestic and farmer's HP and antigens derived from microbiological culture and extracts of various argan products. Subjects with episodes of flu-like syndrome several hours after handling argan cakes, were submitted to a one-hour challenge to argan cakes followed by physical examination, determination of Carbon Monoxide Diffusing Capacity (DLCO) and chest CT-scan on day 2, and, when necessary, bronchoalveolar lavage on day 4. RESULTS: Six of the nine workers experienced flu-like symptoms within 8 hours after argan handling. After challenge, two subjects presented a significant decrease of DLCO and alveolitis with mild lymphocytosis, and one presented ground glass opacities. These two patients and another patient presented significant arcs to both granulates and non-sterile powder. No reactivity was observed to sterile argan finished product, antigens derived from argan cultures (various species of Bacillus) and Streptomyces marokkonensis (reported in the literature to contaminate argan roots). CONCLUSIONS: We report the first evidence of hypersensitivity pneumonitis related to argan powder in two patients. This implies preventive measures to reduce their exposure and clinical survey to diagnose early symptoms. As exposure routes are different and antibodies were observed against argan powder and not the sterile form, consumers using argan-based cosmetics should not be concerned.
Assuntos
Alveolite Alérgica Extrínseca/etiologia , Cosméticos/efeitos adversos , Pulmão/diagnóstico por imagem , Doenças Profissionais/etiologia , Sapotaceae/efeitos adversos , Adulto , Alveolite Alérgica Extrínseca/diagnóstico , Alveolite Alérgica Extrínseca/fisiopatologia , Líquido da Lavagem Broncoalveolar/citologia , Feminino , Humanos , Masculino , Doenças Profissionais/diagnóstico , Doenças Profissionais/fisiopatologia , Capacidade de Difusão Pulmonar , Tomografia Computadorizada por Raios XRESUMO
Bird fancier's lung (BFL) is a disease produced by exposure to avian proteins present in droppings, blooms, and serum of a variety of birds. Although serological test results are currently used to confirm clinical diagnosis of the disease, bird species specificity is poorly understood. This study aimed to contribute to a better understanding of the specificity of immunogenic proteins revealed from the droppings of three bird species. Sera from four patients with BFL and two controls without exposure were analyzed by Western blotting with antigens from droppings of two pigeon and budgerigar strains and two hen species. When the antigens from the droppings of the three bird species were compared, the profile of immunogenic proteins was different and there were similarities between strains of the same species. Only one 68-kD protein was common to pigeon and budgerigar droppings, while proteins of 200, 175, 140, 100, and 35 kD were detected as specific in one bird species. These results provide insight to further characterize these proteins, and to design new serological tests specific to different bird species. These tests may help to refine strategies of antigenic exclusion and also to allow a patient compensation in case of BFL of occupational origin.
Assuntos
Pulmão do Criador de Aves/sangue , Pulmão do Criador de Aves/diagnóstico , Pulmão do Criador de Aves/imunologia , Animais , Antígenos/sangue , Antígenos/imunologia , Galinhas/imunologia , Columbidae/imunologia , Eletroforese em Gel de Poliacrilamida , Humanos , Melopsittacus/imunologia , Especificidade da EspécieRESUMO
BACKGROUND: Machine operator's lung (MOL) is a hypersensitivity pneumonitis the diagnosis of which is difficult. Our laboratory previously developed an ELISA test using recombinant antigens from Mycobacterium immunogenum isolated in French plant. The objective was to validate the previous ELISA results with ten new suspected cases from Germany. METHODS: Two serological analyses were performed: ELISA with the six recombinant antigens, and electrosyneresis with crude antigens of M. immunogenum and three other main species isolated from contaminated metalworking fluids. RESULTS: The two recombinant antigens acyl-CoA dehydrogenase and dihydrolipoyl dehydrogenase, combined together, and electrosyneresis are useful in making the diagnosis regardless of the clinical and radiological data. Finally 9 out of the 10 suspected cases were declared as MOL. CONCLUSIONS: Despite the geographical distance, the crude and recombinant antigens produced to investigate the clustered French cases also proved to be useful in diagnosing the suspected cases in Germany.
Assuntos
Alveolite Alérgica Extrínseca/diagnóstico , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Distúrbios Distônicos/diagnóstico , Mycobacterium/imunologia , Acil-CoA Desidrogenase/imunologia , Adulto , Alveolite Alérgica Extrínseca/microbiologia , Contraimunoeletroforese , Di-Hidrolipoamida Desidrogenase/imunologia , Distúrbios Distônicos/microbiologia , Ensaio de Imunoadsorção Enzimática , França , Alemanha , Humanos , Masculino , Metalurgia , Pessoa de Meia-IdadeRESUMO
CONTEXT: Bird fancier's lung (BFL) is the most prevalent form of hypersensitivity pneumonitis (HP) worldwide. The current techniques used for the serological diagnosis of BFL all use crude extracts from feathers, droppings, and blooms as test antigens, which is associated with a lack of standardization and variability of the results. An antigenic protein, immunoglobulin lambda-like polypeptide-1 (IgLL1), isolated from pigeon droppings, was recently identified to be associated with BFL. We used genetic engineering to produce IgLL1 as a recombinant antigen. AIM: We aimed to prospectively validate the use of an automated ELISA based on recombinant IgLL1 protein (r-IgLL1) as the test antigen for the serological diagnosis of BFL. METHODS: Immunoprecipitation (IP) techniques (immunodiffusion (ID), immunoelectrophoresis (IEP)) and ELISA using r-IgLL1 were performed concomitantly over 10 months on 634 sera from patients with a BFL serodiagnosis request. Questionnaires were sent to obtain details on the avian exposure, clinical data, and final diagnosis. Concordance, sensitivity (Se), and specificity (Sp) of the two techniques were compared. RESULTS: In total, 72 completed questionnaires were returned with 18 cases of BFL diagnosed and 54 of non-BFL. The concordance between the ELISA and ID+IEP precipitation techniques was 71%. The combination of immunoprecipitation techniques showed a Se of 78% and a Sp of 67%. The ELISA using r-IgLL1 showed a Se of 89% and a Sp of 91%. The automated r-IgLL1 ELISA test is sufficiently efficient to be used alone for the diagnosis of patients exposed solely to Columbidae. In cases of other avian exposure, the Se and Sp of the r-IgLL1 ELISA used for screening combined with the immunodiffusion test for confirmation were 89% and 93%, respectively. CONCLUSIONS: The automated ELISA using r-IgLL1 is a promising tool for BFL serodiagnosis. Replacing immunodiffusion by the automated ELISA using r-IgLL1 as a screening technique will be the basis of our future strategy for BFL serodiagnosis.
Assuntos
Alveolite Alérgica Extrínseca , Proteínas Aviárias , Pulmão do Criador de Aves , Alveolite Alérgica Extrínseca/diagnóstico , Animais , Antígenos , Ensaio de Imunoadsorção Enzimática , Humanos , Metilcelulose , Testes SorológicosRESUMO
OBJECTIVE: This study aims to search for reliable serological biomarkers allowing the early prediction of cystic echinococcosis (CE) post-operative outcomes. METHODS: We applied immunoprecipitation (IP) of Echinococcus granulosus protoscolex antigens with pediatric CE patients' plasma collected at 1-month and 1-year post-surgery, followed by Liquid Chromatography with tandem mass spectrometry (LC-MS/MS). We compared IP proteomic content from relapsed patients within the first-year post-surgery (RCE) to cases with no relapses until 3 post-operative years (NRCE). Selected proteins were recombinantly synthesized and assessed for their prognostic performance by Enzyme-linked immunosorbent assay (ELISA). RESULTS: A total of 305 immunoreactive parasitic proteins were identified, 59 of which were significantly more abundant in RCE than NRCE for both time-points. Four proteins showed the most promising characteristics for predicting CE outcomes: cytoplasmic malate dehydrogenase (Eg-cMDH), citrate synthase (Eg-CS), annexin A6 and severin. ELISA-IgG against the four markers were significantly lower at 1-year post-surgery than 1-month in NRCE, in contrast to RCE that displayed either stable or higher levels. The Eg-cMDH and Eg-CS showed the best prognostic performance, with respective probabilities of being "relapse-free" of 83% and 81%, if a decrease of IgG levels occurred between 1-month and 1-year post-surgery. CONCLUSION: The Eg-cMDH and Eg-CS are promising biomarkers to predict early CE post-surgical outcomes.
Assuntos
Equinococose , Echinococcus granulosus , Animais , Antígenos de Helmintos , Biomarcadores , Criança , Cromatografia Líquida , Equinococose/diagnóstico , Equinococose/cirurgia , Ensaio de Imunoadsorção Enzimática , Humanos , Proteômica , Espectrometria de Massas em Tandem , Resultado do TratamentoRESUMO
Metalworking fluid-associated hypersensitivity pneumonitis (MWF-HP) is a pulmonary disease caused by inhaling microorganisms present in the metalworking fluids used in the industrial sector. Mycobacterium immunogenum is the main etiological agent. Among the clinical, radiological and biological tools used for diagnosis, serological tests are important. The aim of this study was to identify immunogenic proteins in M. immunogenum and to use recombinant antigens for serological diagnosis of MWF-HP. Immunogenic proteins were detected by two-dimensional Western blot and candidate proteins were identified by mass spectrometry. Recombinant antigens were expressed in Escherichia coli and tested by enzyme-linked immunosorbent assay (ELISA) with the sera of 14 subjects with MWF-HP and 12 asymptomatic controls exposed to M. immunogenum. From the 350 spots visualized by two-dimensional gel electrophoresis with M. immunogenum extract, 6 immunogenic proteins were selected to be expressed as recombinant antigens. Acyl-CoA dehydrogenase antigen allowed for the best discrimination of MWF-HP cases against controls with an area under the receiver operating characteristics (ROC) curve of 0.930 (95% CI=0.820-1), a sensitivity of 100% and a specificity of 83% for the optimum threshold. Other recombinant antigens correspond to acyl-CoA dehydrogenase FadE, cytosol aminopeptidase, dihydrolipoyl dehydrogenase, serine hydroxymethyltransferase and superoxide dismutase. This is the first time that recombinant antigens have been used for the serodiagnosis of hypersensitivity pneumonitis. The availability of recombinant antigens makes it possible to develop standardized serological tests which in turn could simplify diagnosis, thus making it less invasive.
Assuntos
Alveolite Alérgica Extrínseca/diagnóstico , Antígenos de Bactérias/sangue , Técnicas de Laboratório Clínico/métodos , Mycobacterium/imunologia , Doenças Profissionais/diagnóstico , Proteínas de Bactérias/análise , Proteínas de Bactérias/imunologia , Clonagem Molecular , Eletroforese em Gel Bidimensional , Escherichia coli/genética , Expressão Gênica , Humanos , Proteoma/análise , Curva ROC , Proteínas Recombinantes/isolamento & purificação , Sensibilidade e Especificidade , Testes Sorológicos/métodosRESUMO
BACKGROUND: Cystic echinococcosis (CE) affects predominantly young patients in highly endemic areas. Improved serological methods are needed for the follow-up of CE cases, especially given the high rates of post-surgical relapse that require detection as soon as possible. METHODS: We designed a study to investigate the value of antigenic proteins extracted from Echinococcus granulosus (E. granulosus) protoscoleces, and of recombinant B2t and 2B2t proteins, for assessing the efficacy of surgical treatment carried out on CE-affected children. This study was performed on 278 plasma samples collected from 59 Tunisian children surgically treated for CE and monitored for 3 years post-surgery. The patients were classified according to post-surgical outcomes into a "non-relapsed" (NRCE) and a "relapsed" (RCE) group. We performed in-house ELISAs to measure anti-B2t and anti-2B2t IgG and immunoblotting for the detection of IgG against SDS-PAGE-resolved E. granulosus protoscoleces-specific antigens. The Wilcoxon test was applied to assess anti-B2t and anti-2B2t IgG levels. We applied the Cochran Q test to compare the distribution of immunoblotting antigenic bands between 1-month and 1-year post-surgery. RESULTS: The probability of being "relapse-free" when a decrease in antibody titers occurred between 1 month and 1 year post-surgery was 81% and 75%, respectively, for anti-B2t and anti-2B2t IgG. We identified five protoscolex protein bands of 20, 26/27, 30, 40 and 46 kDa as highly immunoreactive by immunoblot for both RCE and NRCE patients at 1 month post-surgery, and significantly lower immunoreactivity after 1 year (p < 10-4) for NRCE compared to RCE patients. The proteins at 26/27 and 40 kDa displayed the best performance in predicting the outcome, with an 84% probability of being relapse-free when the reactivity against the 40 kDa antigen, the doublet at 26/27 kDa, or both was absent or disappeared between 1 month and 1 year post-surgery, and a 93% probability of being relapsed when both bands remained reactive or increased in intensity between the two time points. CONCLUSIONS: The B2t protein could be useful for the prediction of CE early post-surgical outcomes. The proteins of E. granulosus protoscoleces, especially the doublet P26/27 and P40, could be promising predictive biomarkers for the post-surgical follow-up of CE cases as well.
Assuntos
Antígenos de Helmintos/sangue , Western Blotting/métodos , Equinococose/sangue , Equinococose/diagnóstico , Echinococcus granulosus/química , Cirurgia Geral , Proteínas de Helminto/sangue , Adolescente , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/imunologia , Criança , Pré-Escolar , Echinococcus granulosus/genética , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Imunoglobulina G/sangue , Masculino , Estudos Prospectivos , Recidiva , Testes Sorológicos/métodos , Resultado do Tratamento , TunísiaRESUMO
This study investigated the plant features associated with increased irritation symptoms and levels of inflammation markers among compost workers (CWs). Ninety CWs were followed over 18 months, using questionnaires on respiratory symptoms, fractional exhaled nitric oxide measurements, spirometry, a methacholine bronchial challenge test, and quantification of specific immunoglobulins E (IgE) and G. CWs in plants processing the highest quantities of waste exhibited more airway irritation symptoms. So did the CWs in partially and fully indoor plants as compared to those in plants entirely outdoors. Working in sewage sludge versus green waste plants and having a high level of exposure were associated with higher levels of different IgE. The duration of employment decreased the FEV1 by 16 ml per year. Working in an indoor plant is linked to symptoms and inflammation markers in CWs.
Assuntos
Poluentes Ocupacionais do Ar/efeitos adversos , Hiper-Reatividade Brônquica/etiologia , Compostagem , Exposição Ocupacional , Plantas , Adulto , Testes de Provocação Brônquica , Humanos , Masculino , Pessoa de Meia-Idade , Espirometria , Inquéritos e QuestionáriosRESUMO
We propose a strategy for serodiagnosis of hypersensitivity pneumonitis (HP): 1) question patients about their private or occupational activity, or visit him on site; 2) select panels of six somatic specific antigens appropriate for each type of exposure; 3) and use ELISA to test concomitantly two recombinant antigens highly specific to Farmer's lung, Metalworking-fluid HP, and for Bird fancier's lung. The serodiagnosis provides an immunological argument that may complete radiological, functional lung exploration and clinical features; 4) If the serodiagnosis is negative but the suspicion of HP is strong, a microbial analysis of the patient's specific exposure is conducted; 5) "A la carte" antigens are produced from the microorganisms isolated in the patient's environment sample and tested; 6) Finally, the patient may be asked to undergo a specific inhalation challenge with the offending antigens in a safety cabin, or to avoid his usual environment for a few days.
Assuntos
Alveolite Alérgica Extrínseca/diagnóstico , Alveolite Alérgica Extrínseca/imunologia , Pulmão do Criador de Aves/imunologia , Testes Sorológicos/métodos , Alveolite Alérgica Extrínseca/diagnóstico por imagem , Alveolite Alérgica Extrínseca/etiologia , Antígenos/imunologia , Testes de Provocação Brônquica/métodos , Testes de Provocação Brônquica/normas , Exposição Ambiental/efeitos adversos , Humanos , Valor Preditivo dos Testes , Radiografia/normas , Inquéritos e Questionários/estatística & dados numéricosRESUMO
The use of recombinant antigens has been shown to improve both the sensitivity and the standardization of the serological diagnosis of Farmer's lung disease (FLD). The aim of this study was to complete the panel of recombinant antigens available for FLD serodiagnosis with antigens of Lichtheimia corymbifera, known to be involved in FLD. L. corymbifera proteins were thus separated by 2D electrophoresis and subjected to western blotting with sera from 7 patients with FLD and 9 healthy exposed controls (HEC). FLD-associated immunoreactive proteins were identified by mass spectrometry based on a protein database specifically created for this study and subsequently produced as recombinant antigens. The ability of recombinant antigens to discriminate patients with FLD from controls was assessed by ELISA performed with sera from FLD patients (n = 41) and controls (n = 43) recruited from five university hospital pneumology departments of France and Switzerland. Forty-one FLD-associated immunoreactive proteins from L. corymbifera were identified. Six of them were produced as recombinant antigens. With a sensitivity and specificity of 81.4 and 77.3% respectively, dihydrolipoyl dehydrogenase was the most effective antigen for discriminating FLD patients from HEC. ELISA performed with the putative proteasome subunit alpha type as an antigen was especially specific (88.6%) and could thus be used for FLD confirmation. The production of recombinant antigens from L. corymbifera represents an additional step towards the development of a standardized ELISA kit for FLD diagnosis.
Assuntos
Antígenos de Fungos/metabolismo , Pulmão de Fazendeiro/diagnóstico , Mucorales/metabolismo , Antígenos de Fungos/classificação , Antígenos de Fungos/genética , Antígenos de Fungos/imunologia , Western Blotting , Estudos de Casos e Controles , Bases de Dados Genéticas , Eletroforese em Gel Bidimensional , Ensaio de Imunoadsorção Enzimática , Pulmão de Fazendeiro/microbiologia , Feminino , Humanos , Imunoglobulina G/sangue , Masculino , Espectrometria de Massas , Mucorales/genética , Mucorales/isolamento & purificação , RNA Fúngico/isolamento & purificação , RNA Fúngico/metabolismo , RNA Mensageiro/química , RNA Mensageiro/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Sensibilidade e Especificidade , Análise de Sequência de RNARESUMO
Very few studies have been conducted on cystic fibrosis (CF) patients' exposure to the indoor environment and, to our knowledge, there are no studies dealing with the link between specific fungal environmental exposure at home and fungal colonization resulting in allergic bronchopulmonary aspergillosis (ABPA). Fungal exposure of CF adult patients with ABPA (n=4) with fungal sensitization (n=7) and with no ABPA (n=5) was assessed in 16 homes by dust sampling with electrostatic dust fall collectors (EDCs). Aspergillus fumigatus was specifically quantified by real-time quantitative polymerase chain reactions (qPCRs), and A. fumigatus DNA concentrations were significantly higher in homes of ABPA patients (p<0.001). Results indicate that indoor fungal contamination could be a factor favoring ABPA and suggest that environmental surveys could help in preventing fungal risk in CF patients.
Assuntos
Poluição do Ar em Ambientes Fechados , Aspergilose Broncopulmonar Alérgica , Aspergillus fumigatus/isolamento & purificação , Fibrose Cística , Exposição por Inalação , Adulto , Poluição do Ar em Ambientes Fechados/efeitos adversos , Poluição do Ar em Ambientes Fechados/análise , Poluição do Ar em Ambientes Fechados/prevenção & controle , Antígenos de Fungos/análise , Aspergilose Broncopulmonar Alérgica/complicações , Aspergilose Broncopulmonar Alérgica/diagnóstico , Aspergilose Broncopulmonar Alérgica/microbiologia , Aspergilose Broncopulmonar Alérgica/prevenção & controle , Fibrose Cística/complicações , Fibrose Cística/microbiologia , DNA Fúngico/análise , Monitoramento Ambiental/métodos , Feminino , França , Humanos , Exposição por Inalação/efeitos adversos , Exposição por Inalação/análise , Exposição por Inalação/prevenção & controle , MasculinoRESUMO
Electrosyneresis and double diffusion are immunoprecipitation techniques commonly used in the serological diagnosis of Farmer's lung disease (FLD). These techniques are reliable but lack standardization. The aim of this study was to evaluate Western blotting for the serodiagnosis of FLD. We carried out Western blotting with an antigenic extract of Lichtheimia corymbifera, an important aetiological agent of the disease. The membranes were probed with sera from 21 patients with FLD and 21 healthy exposed controls to examine the IgG antibody responses against purified somatic antigens. Given the low prevalence of the disease, 21 patients could be considered as a relevant series. Four bands were significantly more frequently represented in membranes probed with FLD sera (bands at 27.7, 40.5, 44.0 and 50.5 kDa) than those probed with control sera. We assessed the diagnostic value of different criteria alone or in combination. The diagnostic accuracy of the test was highest with the inclusion of at least two of the following criteria: at least five bands on the strip and the presence of one band at 40.5 or 44.0 kDa. Sensitivity, specificity and positive and negative predictive values were all 81%, and the odds ratio was 18.06. Inclusion of bands of high intensity diminished rather than improved the diagnostic value of the test. We concluded that Western blotting is a valuable technique for the serodiagnosis of FLD. The industrial production of ready-to-use membranes would enable the routine use of this technique in laboratories, and provide reliable and standardized diagnostic results within a few hours.