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1.
J Proteome Res ; 20(5): 2618-2627, 2021 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-33823594

RESUMO

New approaches to rapid, simple, in vitro diagnostic immunoassays that do not rely on centralized laboratory facilities are urgently needed for disease diagnosis and to inform treatment strategies. The recent and ongoing COVID-19 pandemic has emphasized that rapid diagnostics are needed to help guide government policies on quarantines, social distancing measures, and community lockdowns. A common approach to developing new immunoassays is to modify existing platforms (e.g., automated ELISA and lateral flow assays) for the new analyte, even though this does not address the drawbacks of existing platforms. An alternate approach is to search for robust assays that have been superseded but could in fact solve important challenges using modern technologies. Immunodiffusion is one such platform based on unique "precipitin ring" patterns formed in gels or paper following interactions between proteins and cognate antibodies in diffusion/reaction systems. Herein, we investigate the microstructure of these precipitin rings using a combination of fluorescence and electron microscopy and also perform a mass spectrometry investigation to determine the proteomic composition of the rings. We observed that the rings were composed of microparticles, which we termed "precipitin complexes", and that these complexes were composed of at least 19 key proteins, including immunoglobulins and complement factors along with a range of plasma proteins, possibly related to immune complexes and/or high-density lipoprotein particles. This information will be useful in developing new in vitro diagnostics using reaction/diffusion systems-techniques that require a single assay step and that only require calibrated length measurements for target protein quantification.


Assuntos
COVID-19 , Proteômica , Controle de Doenças Transmissíveis , Humanos , Imunodifusão , Microscopia , Pandemias , Precipitinas , SARS-CoV-2
2.
PLoS One ; 14(8): e0221228, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31412087

RESUMO

Early and accurate diagnosis of coccidioidomycosis, also known as Valley fever, is critical for appropriate disease treatment and management. Current serodiagnosis is based on the detection of patient serum antibodies that react with tube precipitin (TP) and complement fixation (CF) antigens of Coccidioides. IgM is the first class of antibodies produced by hosts in response to coccidioidal insults. The highly glycosylated ß-glucosidase 2 (BGL2) is a major active component of the TP antigen that stimulates IgM antibody responses during early Coccidioides infection. The predominant IgM epitope on BGL2 is a unique 3-O-methyl-mannose moiety that is not produced by commonly used protein expression systems. We genetically engineered and expressed a recombinant BGL2 (rBGL2ur), derived from Coccidioides, in non-pathogenic Uncinocarpus reesii, a fungus phylogenetically related to the Coccidioides pathogen. The rBGL2ur protein was purified from the culture medium of transformed U. reesii by nickel affinity chromatography, and the presence of 3-O-methyl mannose was demonstrated by gas chromatography. Seroreactivity of the purified rBGL2ur protein was tested by enzyme-linked immunosorbent assays using sera from 90 patients with coccidioidomycosis and 134 control individuals. The sensitivity and specificity of the assay with rBGL2ur were 78.8% and 87.3%, respectively. These results were comparable to those obtained using a proprietary MiraVista Diagnostic (MVD) IgM (63.3% sensitivity; 96.3% specificity), but significantly better than the ID-TP assay using non-concentrated patient sera (33.3% sensitivity; 100% specificity). Expression of rBGL2ur in U. reesii retains its antigenicity for coccidioidomycosis serodiagnosis and greatly reduces biosafety concerns for antigen production, as Coccidioides spp. are biological safety level 3 agents.


Assuntos
Anticorpos Antifúngicos , Antígenos de Fungos/imunologia , Coccidioides , Coccidioidomicose , Precipitinas , Saccharomycetales , Testes Sorológicos , Anticorpos Antifúngicos/química , Anticorpos Antifúngicos/imunologia , Coccidioides/química , Coccidioides/genética , Coccidioides/imunologia , Coccidioidomicose/diagnóstico , Coccidioidomicose/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Precipitinas/química , Precipitinas/imunologia , Saccharomycetales/química , Saccharomycetales/genética
3.
Allergol Int ; 67S: S12-S17, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29773475

RESUMO

BACKGROUND: Several serological tests for specific precipitin or IgG are available to demonstrate type III hypersensitivity reactions to Aspergillus species and are essential for infectious fungal disease diagnosis. These assays are also important for allergic bronchopulmonary aspergillosis (ABPA) diagnosis; however, their concordance in ABPA was not well studied. METHODS: Fifty-two ABPA patients diagnosed based on ISHAM criteria were enrolled. Precipitins and IgG specific to Aspergillus fumigatus, Aspergillus niger, Aspergillus flavus, or Aspergillus terreus were measured using Ouchterlony double immunodiffusion tests and ImmunoCAP method, respectively. A. fumigatus-specific IgG was also determined using complement-fixation (CF) method. RESULTS: Forty-eight percent of cases were double-positive for A. fumigatus-specific precipitin and IgG (ImmunoCAP), whereas 3 (6%) and 14 (28%) cases were positive for precipitin or IgG alone, respectively. Kappa coefficient between these measurements was 0.32, suggesting poor concordance. Double-positive cases were more likely to present: Aspergillus sp. in sputum culture, lower pulmonary functions, peripheral blood eosinophilia, higher total IgE and A. fumigatus-specific IgG titer than precipitin-negative cases. A. fumigatus-specific IgG (CF) was positive only in 8 (15%) cases. The presence of A. fumigatus-specific precipitin or IgG was associated with antibodies specific for other Aspergillus spp., suggesting cross-reactivity. CONCLUSIONS: Positive rate of A. fumigatus-specific precipitin or IgG (ImmunoCAP) was superior to IgG (CF), but relatively poor concordance was noted between precipitin and IgG (ImmunoCAP). Positive precipitin for A. fumigatus suggests more active diseases. Cross-reactivity may exist between antibodies to different Aspergillus spp. Therefore, the type III hypersensitivity results in ABPA diagnosis should be carefully evaluated.


Assuntos
Anticorpos Antifúngicos/imunologia , Aspergilose Broncopulmonar Alérgica/imunologia , Aspergillus/imunologia , Imunoglobulina G/imunologia , Precipitinas/imunologia , Idoso , Feminino , Humanos , Imunoglobulina E/imunologia , Masculino , Pessoa de Meia-Idade , Escarro/microbiologia
4.
J Vet Diagn Invest ; 30(1): 42-55, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28906178

RESUMO

The gel diffusion precipitin test (GDPT) and restriction endonuclease analysis (REA) have commonly been used in the serotyping and genotyping of Pasteurella multocida. Whole genome sequencing (WGS) and single nucleotide polymorphism (SNP) analysis has become the gold standard for other organisms, offering higher resolution than previously available methods. We compared WGS to REA and GDPT on 163 isolates of P. multocida to determine if WGS produced more precise results. The isolates used represented the 16 reference serovars, isolates with REA profiles matching an attenuated fowl cholera vaccine strain, and isolates from 10 different animal species. Isolates originated from across the United States and from Chile. Identical REA profiles clustered together in the phylogenetic tree. REA profiles that differed by only a few bands had fewer SNP differences than REA profiles with more differences, as expected. The GDPT results were diverse but it was common to see a single serovar show up repeatedly within clusters. Several errors were found when examining the REA profiles. WGS was able to confirm these errors and compensate for the subjectivity in analysis of REA. Also, results of WGS and SNP analysis correlated more closely with the epidemiologic data than GDPT. In silico results were also compared to a lipopolysaccharide rapid multiplex PCR test. From the data produced in our study, WGS and SNP analysis was superior to REA and GDPT and highlighted some of the issues with the older tests.


Assuntos
Pasteurella multocida/isolamento & purificação , Mapeamento por Restrição/veterinária , Sorotipagem/veterinária , Sequenciamento Completo do Genoma/veterinária , Animais , Proteínas de Bactérias/análise , Enzimas de Restrição do DNA/análise , DNA Bacteriano/análise , Imunodifusão/métodos , Imunodifusão/veterinária , Infecções por Pasteurella/veterinária , Filogenia , Precipitinas/química , Mapeamento por Restrição/métodos , Sorotipagem/métodos , Sequenciamento Completo do Genoma/métodos
5.
Eye (Lond) ; 30(1): 127-32, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26493031

RESUMO

PURPOSE The number of antigen-presenting cells decreases during organ culture of corneoscleral discs. This might result in a decrease of immune reactions with increasing duration of organ culture. To investigate this hypothesis, we performed a retrospective analysis of all penetrating keratoplasties that were consecutively performed over the last 5 years.PATIENTS AND METHODS All cases of penetrating keratoplasties (n=1006) were divided into two groups, with the division made at the median of the storage time (21 days). These two groups were compared by a Cox proportional hazards survival model regarding the incidence of endothelial immune reactions, clear graft survival, and chronic endothelial cell loss following penetrating keratoplasty considering patient's age, donor's age, and risk situation as co-variates.RESULTS We observed statistically significantly fewer endothelial immune reactions (20.1% (95% confidence interval 15.5-24.5%) after 2 years) in the group with a storage time of more than 21 days compared with the group with a storage time of <21 days (26.5% (95% confidence interval 21.6-31.2%) after 2 years). However, the duration of organ culture did not have a statistically significant effect on clear graft survival or chronic endothelial cell loss.CONCLUSION Our results demonstrate that an increased duration of organ culture leads to a lower incidence of endothelial immune reactions following penetrating keratoplasty. However, we do not recommend increased storage times in general as overall graft survival did not improve. The reason for this apparent paradox may be that the endothelial cell count decreases during storage time.


Assuntos
Células Apresentadoras de Antígenos/imunologia , Endotélio Corneano/imunologia , Ceratoplastia Penetrante , Adulto , Idoso , Perda de Células Endoteliais da Córnea/patologia , Criopreservação/métodos , Sobrevivência de Enxerto/fisiologia , Humanos , Incidência , Pessoa de Meia-Idade , Técnicas de Cultura de Órgãos , Preservação de Órgãos/métodos , Precipitinas/sangue , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Fatores de Tempo , Doadores de Tecidos
6.
Pathology ; 47(1): 62-7, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25474508

RESUMO

Unidentified precipitin lines (UPLs) are lines in immune precipitin assays which do not characterise with known extractable nuclear antigen (ENA) antibodies. Currently, the clinical significance of UPLs is uncertain.The aim of this study was to determine the clinical and laboratory correlates of UPLs detected over a 3 year period in a regional immunopathology laboratory.A total of 144 patients with UPLs on ENA testing were retrospectively analysed.ENA by counterimmunoelectrophoresis (CIE) was performed on 11,869 patient samples with further characterisation being performed for 1437 positive results. Ten percent of the positive ENAs demonstrated UPLs. The majority of patients with UPLs were female (71%) with an average age of 60 years. Precipitin lines for both continuously growing myeloid cell line K562 and rabbit thymus extract (RTE) were more frequent (47%), compared to K562 only (41%) or RTE only (12%). The most common antinuclear antibody (ANA) patterns associated with UPLs were speckled (29%), homogenous (16%), mixed patterns (14%), with 30% negative ANA and 5% showing cytoplasmic patterns (Golgi, GWB, mitochondrial). Both ANA positive and ANA negative patients with UPLs were generally associated with connective tissue and autoimmune disease with the majority demonstrating a positive association of UPLs with Ro52 and/or Ro60/SSA detected by Euroimmun line immunoassay. UPLs frequently seen with a negative ANA were also identified in renal disease, pulmonary fibrosis/bronchiectasis and malignancy/lymphoma.UPLs have uncertain diagnostic utility at this stage and further work needs to be done to clarify this question. UPLs were found in systemic and organ specific autoimmune disease, renal, pulmonary and neoplastic disorders. UPLs can occur in both ANA positive and ANA negative sera. The nature of the precipitating antigen in UPLs is still obscure.


Assuntos
Anticorpos Antinucleares/análise , Antígenos Nucleares/imunologia , Doenças do Tecido Conjuntivo/diagnóstico , Imunoprecipitação , Precipitinas , Doenças Autoimunes/diagnóstico , Doenças do Tecido Conjuntivo/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
7.
J. investig. allergol. clin. immunol ; 25(4): 237-250, 2015. tab, ilus
Artigo em Inglês | IBECS | ID: ibc-138419

RESUMO

Hypersensitivity pneumonitis (HP), also known as extrinsic allergic alveolitis, is a complex pulmonary syndrome mediated by the immune system and caused by inhalation of a wide variety of antigens to which the individual has been previously sensitized. The pathobiology of the disease is not fully understood, but in addition to the triggers that initiate the disease, host/genetic factors are likely to be important, as only a minority of exposed individuals develop HP. Due to the lack of a diagnostic gold standard, the diagnosis of HP is not straightforward and relies on the integration of a number of factors, including history of exposure, precipitating antibodies to the offending antigen, clinical features, bronchoalveolar lavage, and radiological and pathologic features. However, in the appropriate setting, a high index of suspicion is critically important and may obviate the need for more invasive tests. Clinical presentation and natural history vary widely. Acute forms generally resolve without sequelae, while chronic forms, which are caused by persistent low-grade exposures, are associated with poor prognosis. Corticosteroids may be useful in acute episodes for symptomatic relief or in chronic and progressive disease, but their long-term efficacy has never been validated in prospective clinical trials. Ideally, patients with HP should be referred to centers with expertise, as the overlap with other forms of interstitial lung disease may be substantial. Making the correct diagnosis has critical therapeutic and prognostic implications (AU)


La neumonitis por hipersensibilidad (NH), también conocida como alveolitis alérgica extrínseca, es un síndrome pulmonar complejo mediado por el sistema inmune y provocado por la inhalación de una amplia variedad de alérgenos frente a los cuales el paciente está previamente sensibilizado. La patogénesis de la enfermedad se conoce parcialmente; sin embargo, además de los activadores iniciales que dan lugar al desarrollo de la enfermedad hay factores genéticos que tienen importancia también, de tal forma que solo una minoría de los individuos expuestos desarrollan la enfermedad. Debido a la falta de un 'gold estándar' para el diagnóstico de NH, se hace necesaria la integración de un número de factores, entre los que se encuentran la historia de exposición al alérgeno, la presencia de anticuerpos precipitantes frente al Ag ofensor, datos clínicos y datos patológicos en el lavado broncoalveolar, y radiológicos. En cualquier caso un alto índice de sospecha clínica es crítica y puede obviar la necesidad de otros test más invasivos. La presentación clínica y la historia natural de la enfermedad puede variar ampliamente desde las formas agudas que generalmente se resuelven sin secuelas a las formas crónicas fibróticas que son provocadas por la exposición de grado bajo mantenida y que se asocian con un peor pronóstico. Los corticosteroides pueden ser útiles en el tratamiento sintomático de los episodios agudos o en la enfermedad crónica progresiva, pero su eficacia a largo plazo nunca ha sido validada en ensayos clínicos diseñados para ese fin. La dinámica adecuada debe dirigir a los pacientes con MH a centros especializados expertos, dado que puede haber otras formas solapadas de enfermedad pulmonar y el diagnóstico correcto es crítico para la aplicación de un correcto tratamiento y un mejor pronóstico (AU)


Assuntos
Feminino , Humanos , Masculino , Alveolite Alérgica Extrínseca/epidemiologia , Alveolite Alérgica Extrínseca/etiologia , Reações Antígeno-Anticorpo/imunologia , Fibrose Pulmonar Idiopática/diagnóstico , Doenças Pulmonares Intersticiais/fisiopatologia , Diagnóstico Diferencial , Fibrose Pulmonar Idiopática/etiologia , Prognóstico , Linfócitos/patologia , Imunoglobulina G/isolamento & purificação , Precipitinas/isolamento & purificação , Broncoscopia/tendências , Lavagem Broncoalveolar
8.
Cornea ; 33(4): 331-4, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24488131

RESUMO

PURPOSE: The migratory capacity of donor corneal endothelial cells after Descemet membrane endothelial keratoplasty (DMEK) is not fully understood. We report 2 patients who developed immune reactions after DMEK with endothelial precipitates detectable not only on the grafts but also on host corneal areas stripped off the host Descemet membrane during surgery and initially not covered by the donor Descemet membrane and endothelium ("naked stroma"), strongly suggesting that migration of donor-derived endothelial cells had occurred. METHODS: Observational case series of 2 patients. RESULTS: A 71-year-old man (case 1) and an 84-year-old man (case 2) with Fuchs endothelial dystrophy underwent successful DMEK surgery. Postoperatively, the peripheral corneal stroma showed a denuded area that was not covered by the DMEK graft or by the patients' residual Descemet membrane (because of large descemetorhexis and slight graft decentration). After 18 (case 1) and 6 (case 2) months, a diffuse endothelial immune reaction with precipitates on the graft and, surprisingly, also on peripheral corneal areas that were initially denuded of the host Descemet membrane and not covered by the donor Descemet membrane was observed. The outermost corneal parts covered by the patients' own residual Descemet membrane showed no precipitates. Under treatment with topical corticosteroids, the precipitates rapidly disappeared. Visual acuity, central corneal thickness, and endothelial cell counts were not significantly affected. CONCLUSIONS: The immune reaction episodes in our patients with endothelial precipitates detectable on adjacent host areas initially stripped off and not covered by donor Descemet membrane provide direct in vivo evidence of donor corneal endothelial cell migration after DMEK, filling areas of "naked stroma."


Assuntos
Movimento Celular/fisiologia , Ceratoplastia Endotelial com Remoção da Lâmina Limitante Posterior , Endotélio Corneano/fisiologia , Distrofia Endotelial de Fuchs/cirurgia , Precipitinas , Idoso , Idoso de 80 Anos ou mais , Contagem de Células , Glucocorticoides/uso terapêutico , Humanos , Implante de Lente Intraocular , Masculino , Facoemulsificação , Prednisolona/análogos & derivados , Prednisolona/uso terapêutico , Doadores de Tecidos , Acuidade Visual
10.
Med Mycol ; 50(6): 654-7, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22329454

RESUMO

Hypersensitivity pneumonitis (HP) is a pulmonary granulomatosis involving an immunoallergic mechanism caused by chronic inhalation of antigens, most frequently organic substances, as well as chemicals. We report the first European case of hypersensitivity pneumonitis due to the inhalation of Shiitake mushroom spores. A 37-year-old French Caucasian man with a one-month history of persistent dry cough, shortness of breath and loss of weight was admitted to our hospital on December 2010. Anamnesis showed he was involved in mushroom production beginning in the summer of 2010. His temperature on admission was 36.6°C and he had a normal blood pressure (135/90 mmHg). Bilateral fine crackles were audible in the base of both lungs. Pulmonary function tests showed a mild restrictive pattern with decreased DLco and a PaO(2) of 65 mmHg, Chest CT scan revealed reticulo-nodular shadows, slight ground glass opacities, liner atelectasis, and subpleural opacities in both lung fields. Bronchoscopy was normal but cytological examination of BAL revealed a predominant lymphocytosis (55%). Serum precipitins to the Shiitake mushroom spores were positive (3 precipitins arcs with high intensity) and as a result we advised the patient to cease his mushroom production activities. The diagnosis of hypersensitivity pneumonitis due to inhalation of Shiitake mushroom spores was established as a result of the improvement of all of his clinical symptoms, i.e., cough, weight loss, bilateral fine crackles, mild restrictive pattern of pulmonary function, and reticulo-nodular shadows on chest CT, once exposure was eliminated. Recent interest in exotic mushrooms varieties, e.g., Shiitake, in developed countries because of their possible medicinal properties might increase the potential risk of HP among mushrooms workers. Therefore, healthcare professionals have to take this new potential respiratory disease into account.


Assuntos
Alveolite Alérgica Extrínseca/microbiologia , Exposição por Inalação/efeitos adversos , Cogumelos Shiitake/imunologia , Esporos Fúngicos/imunologia , Adulto , Alveolite Alérgica Extrínseca/imunologia , Antígenos de Fungos/efeitos adversos , Antígenos de Fungos/imunologia , Líquido da Lavagem Broncoalveolar/imunologia , Broncoscopia , Humanos , Pulmão/microbiologia , Pulmão/patologia , Linfocitose/imunologia , Linfocitose/microbiologia , Masculino , Doenças Profissionais/imunologia , Doenças Profissionais/microbiologia , Testes de Precipitina , Precipitinas/sangue , Testes de Função Respiratória , Tórax/diagnóstico por imagem , Tomografia Computadorizada por Raios X , Ultrassonografia
11.
Am J Ophthalmol ; 152(3): 449-453.e1, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21676373

RESUMO

PURPOSE: To describe immune ring (IR) formation as a manifestation of cytomegalovirus (CMV) endotheliitis. DESIGN: Retrospective observational case series. METHODS: Review of all consecutive CMV anterior uveitis and endotheliitis patients seen at the Singapore National Eye Centre for the occurrence of an IR. CMV infection was diagnosed by polymerase chain reaction analysis of the aqueous. RESULTS: None of the 72 eyes with CMV anterior uveitis had an IR formation. Four episodes of IR formation were seen in 3 eyes (14.3%) of 21 cases of CMV-positive endotheliitis. All were unilateral and all 3 patients were human immunodeficiency virus negative middle-aged Chinese males. The IR developed 2 to 7 months after occurrence of the endotheliitis and the aqueous was positive for CMV during 2 of the episodes of IR formation. In 2 eyes, the IR occurred at the completion of a course of systemic ganciclovir when the aqueous was negative for CMV. The immune rings resolved with combination therapy of topical prednisolone acetate 0.12% and ganciclovir. Patient 1 had a recurrence of the IR 4 months after stopping treatment but again resolved following treatment with ganciclovir and topical corticosteroids. CONCLUSION: Corneal immune rings can occur as a result of CMV infection. Hence CMV infection may have to be considered in such cases.


Assuntos
Infecções por Citomegalovirus/diagnóstico , Endotélio Corneano/imunologia , Infecções Oculares Virais/diagnóstico , Uveíte Anterior/diagnóstico , Administração Tópica , Adulto , Antivirais/administração & dosagem , Antivirais/uso terapêutico , Humor Aquoso/virologia , Citomegalovirus/genética , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/tratamento farmacológico , Infecções por Citomegalovirus/imunologia , DNA Viral/análise , Quimioterapia Combinada , Infecções Oculares Virais/tratamento farmacológico , Infecções Oculares Virais/imunologia , Feminino , Ganciclovir/administração & dosagem , Ganciclovir/uso terapêutico , Glucocorticoides/administração & dosagem , Glucocorticoides/uso terapêutico , Soronegatividade para HIV , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Precipitinas/imunologia , Prednisolona/administração & dosagem , Prednisolona/análogos & derivados , Prednisolona/uso terapêutico , Estudos Retrospectivos , Resultado do Tratamento , Uveíte Anterior/tratamento farmacológico , Uveíte Anterior/imunologia
12.
Eur Respir J ; 37(3): 640-7, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20693254

RESUMO

Metalworking fluids (MWF) are responsible for hypersensitivity pneumonitis (HP). The aim of the present study was to identify the antigen (Ag) responsible for MWF-associated HP, and to optimise serological diagnosis by definition of a threshold allowing discrimination between HP patients and asymptomatic exposed workers. 13 patients, who were workers at a car engine manufacturing plant, were suspected of MWF-associated HP. Microbial analysis of 83 used MWFs was carried out. Sera from 13 MWF-associated HP patients, 12 asymptomatic exposed workers and 18 healthy unexposed controls were tested to determine their immunological responses to three Ags, including Mycobacterium immunogenum. M. immunogenum was identified in 40% of used fluids by culture and confirmed by DNA sequencing. The threshold for differentiating MWF-associated HP patients from asymptomatic exposed workers was five arcs of precipitation (sensitivity 77% and specificity 92%), as determined by electrosyneresis (ES). Using ELISA methods with protein extract from M. immunogenum, a threshold leading to 92% sensitivity and 100% specificity was established. The detection of specific antibodies against M. immunogenum Ag at high levels in case sera suggests that M. immunogenum-contaminated MWF is responsible for MWF-associated HP. To discriminate MWF-associated HP patients from asymptomatic exposed workers, we suggest a five-arc threshold for ES and a 1.6-AU threshold for ELISA methods.


Assuntos
Alveolite Alérgica Extrínseca/microbiologia , Óleos Industriais/microbiologia , Mycobacterium/metabolismo , Doenças Profissionais/microbiologia , Adulto , Alveolite Alérgica Extrínseca/patologia , Celulose/análogos & derivados , Celulose/química , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Hipersensibilidade , Masculino , Metalurgia , Pessoa de Meia-Idade , Doenças Profissionais/diagnóstico , Exposição Ocupacional , Precipitinas/química , Análise de Sequência de DNA
13.
Korean J Lab Med ; 30(4): 334-8, 2010 Aug.
Artigo em Coreano | MEDLINE | ID: mdl-20805703

RESUMO

Involvement of the central nervous system is very uncommon in multiple myeloma, observed in approximately 1% of the multiple myeloma patients. We report a case of central nervous system myelomatosis with complex chromosome aberrations in a 62-yr-old female patient, who had previously been diagnosed as multiple myeloma. Fluorescent in situ hybridization revealed 13q deletion, p53 gene deletion and IGH/FGFR3 rearrangement and chromosomal study showed complex chromosome aberrations. After four cycles of chemotherapy, the patient was admitted to the hematology department with severe headache. Plasma cells were found in the cerebrospinal fluid (CSF), and CSF immunoelectrophoresis revealed abnormal precipitin arcs against anti-IgG and anti-lambda antisera. She was given systemic chemotherapy and eight courses of intrathecal chemotherapy, which cleared plasma cells in the CSF. Two months later, she was given autologous stem cell transplantation. Three months after stem cell transplantation, central nervous system myelomatosis progressed to plasma cell leukemia and two months later, the patient expired.


Assuntos
Neoplasias do Sistema Nervoso Central/diagnóstico , Deleção Cromossômica , Mieloma Múltiplo/diagnóstico , Translocação Genética , Antineoplásicos/uso terapêutico , Neoplasias do Sistema Nervoso Central/tratamento farmacológico , Neoplasias do Sistema Nervoso Central/genética , Líquido Cefalorraquidiano/citologia , Terapia Combinada , Progressão da Doença , Feminino , Deleção de Genes , Humanos , Imunoeletroforese , Hibridização in Situ Fluorescente , Leucemia Plasmocitária/diagnóstico , Pessoa de Meia-Idade , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/genética , Plasmócitos/patologia , Precipitinas/metabolismo , Receptor Tipo 3 de Fator de Crescimento de Fibroblastos/genética , Transplante de Células-Tronco , Transplante Autólogo , Proteína Supressora de Tumor p53/genética
14.
Clin Vaccine Immunol ; 17(9): 1330-6, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20631331

RESUMO

Allergic bronchopulmonary aspergillosis (ABPA) is a frequent complication in cystic fibrosis patients. The diagnosis remains difficult and requires a combination of clinical, radiological, biological, and mycological criteria. The aim of this study was to analyze the added value of two recombinant antigens, rAspf4 and rAspf6, associated with the detection of specific IgG; precipitins; total IgE; and Aspergillus fumigatus in sputum for the diagnosis of ABPA. In a retrospective study, we determined the specific IgE responses to these recombinants in 133 sera of 65 cystic fibrosis patients. We selected an average of five serum samples from each of the 17 patients with ABPA (13 proven and 4 probable ABPA) and from 3 patients with Aspergillus bronchitis and rhinosinusitis. One serum sample for the 45 patients without ABPA was tested. The sensitivity of specific IgE detection against rAspf4 calculated per patient (92.3%) was significantly higher (P < 0.05) than that of rAspf6 (53.8%). When rAspf4 IgE detection was associated with anti-Aspergillus IgG enzyme-linked immunosorbent assay (ELISA) and precipitin detection, the sensitivity rose to 100%. The specificities of rAspf4 and rAspf6 IgE detection were 93.7% and 91.6%, respectively. Other diagnostic criteria had slightly lower specificities (87.5% for anti-Aspergillus IgG ELISA, 89.6% for precipitins, 84.4% for total IgE, and 85.0% for positive A. fumigatus culture in sputum). In conclusion, this retrospective study showed the relevance of rAspf4 IgE detection, in combination with other biological markers (Aspergillus IgG ELISA, precipitins, and total IgE), for improving the biological diagnosis of ABPA.


Assuntos
Alérgenos , Aspergilose Broncopulmonar Alérgica/diagnóstico , Biomarcadores , Fibrose Cística/complicações , Micologia/métodos , Adolescente , Adulto , Anticorpos Antifúngicos/sangue , Aspergillus fumigatus/isolamento & purificação , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Lactente , Masculino , Pessoa de Meia-Idade , Precipitinas/sangue , Proteínas Recombinantes , Estudos Retrospectivos , Sensibilidade e Especificidade , Escarro/microbiologia , Adulto Jovem
15.
J Asthma ; 47(1): 2-6, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20100013

RESUMO

There is a substantial body of evidence supporting an association between asthma severity and fungal exposure and sensitization. Fungal allergens are a recognized risk factor for severe asthma. We describe the case of a 44-year-old asthmatic whose asthma control deteriorated after moving to a new flat with walls covered in mould. Allergic bronchopulmonary aspergillosis was excluded. Although sensitization to Candida was demonstrated by a positive Candida-specific radioallergosorbent test, the patient did not entirely satisfy the criteria for a diagnosis of allergic bronchopulmonary candidiasis. The patient's asthma control improved after engaging in a monthly washing regimen of the walls. This case further demonstrates the association between fungal sensitization and asthma severity. The term severe asthma with fungal sensitization has been recently coined to describe this phenomenon.


Assuntos
Asma/imunologia , Asma/fisiopatologia , Fungos/imunologia , Adulto , Poluição do Ar em Ambientes Fechados/efeitos adversos , Asma/diagnóstico por imagem , Asma/terapia , Candida albicans/imunologia , Feminino , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Precipitinas/sangue , Precipitinas/imunologia , Radiografia
16.
Am J Pathol ; 176(2): 608-18, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20056836

RESUMO

IgA nephropathy (IgAN) and Henoch-Schönlein purpura (HSP) are diseases characterized by IgA deposits in the kidney and/or skin. Both may arise after upper respiratory tract infections, but the pathogenic mechanisms governing these diseases remain unclear. Patients with IgAN (n = 16) and HSP (n = 17) were included in this study aimed at examining whether IgA-binding M proteins of group A streptococci could be involved. As M proteins vary in sequence, the study focused on the IgA-binding-region (IgA-BR) of three different M proteins: M4, M22, and M60. Renal tissue from IgAN and HSP patients and skin from HSP patients were examined for deposits of streptococcal IgA-BR by immunohistochemistry and electron microscopy using specific antibodies, and a skin sample from a HSP patient was examined by mass spectrometry. IgA-BR deposits were detected in 10/16 IgAN kidneys and 7/13 HSP kidneys. Electron microscopy demonstrated deposits of IgA-BRs in the mesangial matrix and glomerular basement membrane, which colocalized with IgA. Skin samples exhibited IgA-BR deposits in 4/5 biopsies, a result confirmed by mass spectrometry in one patient. IgA-BR deposits were not detected in normal kidney and skin samples. Taken together, these results demonstrate IgA-BR from streptococcal M proteins in patient tissues. IgA-BR, would on gaining access to the circulation, encounter circulatory IgA and form a complex with IgA-Fc that could deposit in tissues and contribute to the pathogenesis of IgAN and HSP.


Assuntos
Antígenos de Bactérias/metabolismo , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Transporte/metabolismo , Glomerulonefrite por IGA/metabolismo , Imunoglobulina A/metabolismo , Precipitinas/metabolismo , Adolescente , Adulto , Sequência de Aminoácidos , Antígenos de Bactérias/química , Antígenos de Bactérias/imunologia , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/imunologia , Biópsia , Proteínas de Transporte/química , Proteínas de Transporte/imunologia , Criança , Pré-Escolar , Feminino , Glomerulonefrite por IGA/patologia , Humanos , Imuno-Histoquímica , Rim/metabolismo , Rim/patologia , Masculino , Microscopia Eletrônica , Dados de Sequência Molecular , Precipitinas/ultraestrutura , Ligação Proteica , Pele/metabolismo , Pele/patologia , Pele/ultraestrutura , Adulto Jovem
17.
J Theor Biol ; 264(1): 37-44, 2010 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-20060841

RESUMO

We present the results of our comprehensive study of precipitation pattern formation by interacting immunogenic proteins in a gel medium. Formation of immunoprecipitation patterns was studied both theoretically and experimentally. Based on a system of reaction-diffusion equations, continuous deterministic description provides a quantitative model of reaction kinetics. Discrete stochastic microscopic description was used to supplement the results of reaction-diffusion model by mimicking product aggregation that contributes to a deeper understanding of the mechanism that governs the phenomenon. Our studies have shown that the mechanism of immunoprecipitation pattern formation is specific for protein precipitation and differs from such mechanisms for any inorganic or biological substances. By microscopic examination, we demonstrated that immunoprecipitation patterns can have a microstructure. We found that the microscopic structure of immunoprecipitation patterns results from multicomponent composition of antiserum.


Assuntos
Complexo Antígeno-Anticorpo/química , Reações Antígeno-Anticorpo/imunologia , Géis/química , Imunoprecipitação , Modelos Químicos , Algoritmos , Animais , Anticorpos/química , Anticorpos/imunologia , Anticorpos Monoclonais/química , Anticorpos Monoclonais/imunologia , Diversidade de Anticorpos/imunologia , Simulação por Computador , Humanos , Soros Imunes/imunologia , Imunodifusão , Imunoglobulina G/química , Imunoglobulina G/imunologia , Cinética , Precipitinas/química , Precipitinas/imunologia , Coelhos , Sefarose/química
18.
Ophthalmology ; 117(2): 373-80, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20022116

RESUMO

PURPOSE: To test the hypothesis that morphologic patterns of keratic precipitates (KPs) evaluated by in vivo confocal microscopy (IVCM) can differentiate infectious from noninfectious uveitis. DESIGN: Cross-sectional, observational case series. PARTICIPANTS: Sixty-eight eyes of 53 subjects with uveitis. METHODS: A cross-sectional study was performed in patients with infectious and noninfectious uveitis presenting to a tertiary care eye hospital. Detailed ophthalmologic evaluation was performed in all the subjects. Keratic precipitates were studied by IVCM using the HRT II Rostock corneal module (Heidelberg Engineering GmbH, Heidelberg, Germany) and categorized on the basis of morphologic patterns. MAIN OUTCOME MEASURES: Morphology of KPs by slit-lamp biomicroscopy and confocal microscopy. RESULTS: The age of patients ranged from 15 to 87 years (median 40 years). Thirty-two patients were male (60.37%). Thirty-eight subjects had a unilateral presentation (71.69%) of uveitis. Infectious uveitis was seen in 38 cases (71.69%). The characteristics in KPs as seen in infectious uveitis were dendritic, central globular with dendritic, and infiltrative. In noninfectious uveitis (28.3%), stippled, globular, and multiple globular types of KPs were found. The sensitivity, specificity, and positive predictive value for specific combinations of KPs with an infectious cause were 84.21%, 93.33%, and 96.96%, respectively. CONCLUSIONS: In vivo confocal microscopy can act as an adjunct tool for differentiating infectious from noninfectious uveitis. A central globular with dendritic form of KPs is strongly suggestive of infectious uveitis.


Assuntos
Doenças da Córnea/diagnóstico , Endotélio Corneano/patologia , Infecções Oculares/diagnóstico , Microscopia Confocal , Uveíte/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos Transversais , Reações Falso-Positivas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Precipitinas , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Uveíte/microbiologia , Uveíte/parasitologia , Uveíte/virologia , Adulto Jovem
19.
Br J Radiol ; 82(980): e151-4, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19592397

RESUMO

Allergic bronchopulmonary aspergillosis (ABPA) is a complex immune hypersensitivity reaction to the fungus Aspergillus fumigatus, which occurs in patients with asthma and cystic fibrosis. The common radiological findings include fleeting pulmonary opacities, bronchiectasis and mucoid impaction. Uncommon radiological findings encountered in ABPA include lung masses, perihilar opacities simulating hilar adenopathy and pleural effusions. Herein, we describe a 36-year-old man who presented to the chest clinic with miliary nodules on a chest radiograph and was diagnosed as having ABPA confirmed on both serology and lung biopsy. This is the first report of ABPA presenting as randomly scattered nodules on CT.


Assuntos
Aspergilose Broncopulmonar Alérgica/diagnóstico por imagem , Aspergillus fumigatus/imunologia , Pulmão/diagnóstico por imagem , Adulto , Aspergilose Broncopulmonar Alérgica/sangue , Síndrome de Churg-Strauss/diagnóstico por imagem , Diagnóstico Diferencial , Humanos , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Masculino , Precipitinas/sangue , Testes Cutâneos , Tomografia Computadorizada por Raios X , Tuberculose Miliar/diagnóstico por imagem
20.
J Bras Pneumol ; 35(5): 480-3, 2009 May.
Artigo em Inglês, Português | MEDLINE | ID: mdl-19547859

RESUMO

Pulmonary cavitation is rather uncommon in patients with sarcoidosis, and aspergilloma is even more uncommon in such cases. Here, we present the case of a 63-year-old female patient with cavitary lung disease who had been under treatment for TB for 9 months. A diagnosis of pulmonary sarcoidosis was established based on the fiberoptic bronchoscopy finding of noncaseating granuloma. Treatment with corticosteroids led to a dramatic improvement in symptoms. While under treatment for sarcoidosis, the patient developed an aspergilloma. She presented immediate skin test reactivity to Aspergillus fumigatus, as well as positivity for A. fumigatus serum precipitins. This is the first reported case of aspergilloma formation in a patient with cavitary sarcoidosis in India.


Assuntos
Aspergilose/microbiologia , Aspergillus fumigatus , Pneumopatias Fúngicas/microbiologia , Sarcoidose Pulmonar/complicações , Aspergillus fumigatus/imunologia , Biomarcadores/sangue , Feminino , Humanos , Pessoa de Meia-Idade , Precipitinas/sangue , Sarcoidose Pulmonar/tratamento farmacológico
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