RESUMO
OBJECTIVE: To evaluate the IgE reactivity of sera in patients suffering from type 1 diabetes (T1D), lupus nephritis (LN) and juvenile idiopathic arthritis (JIA) against a molecule constructed from T epitopes of A. lumbricoides allergens. METHODS: We designed and expressed a synthetic multi-epítope protein named MP1 from A. lumbricoides and house dust mites allergens. By indirect ELISA, we evaluated IgE-reactivity to MP1 and to the whole-body extract of Ascaris lumbricoides in 45 sera from Colombian Caribbean patients with lupus nephritis (LN; n=25), type 1 diabetes (T1D; n=10) and Juvenil idiopathic arthritis (JIA; n=10). Individuals with poly autoimmunity were excluded. All patients were referred to the study by their specialist doctor. RESULTS: IgE to whole-body extract of A. lumbricoides showed the following median concentrations.484.2 ng/ml (IQR: 203.4) in JIA patients, 325.6 ng/ml (IQR: 179.3) in individuals with LN, and 424.7 ng/ml (IQR: 80.1) in the T1D group. On the other hand, IgE-reactivity to MP1 was 126.4 ng/ml (IQR: 90.9) in JIA patients, 130.7 ng/ml (IQR: 94.8) in an individual with LN, and 148.8 ng/ml (IQR: 102.1) in the T1D group. Although no statistical differences were observed between patient groups, the IgE to MP1 in all patients (n: 45) (IgE median: 134.2 ng/ml; IQR: 100) were significantly less compared to Ascaris extract (IgE median: 380.7 ng/ml; IQR: 175.8); (W: 0.732; p-value: 1.034x10-7). CONCLUSIONS: These preliminary results suggest that MP1 showed antigenic properties with low IgE- reactivity, compared to Ascaris lumbricoides extracted in individuals with autoimmune diseases. Further studies are needed to understand better the immune response induced by this molecule.
OBJETIVO: Evaluar la reactividad IgE de sueros en pacientes que padecen diabetes tipo 1 (DT1), nefritis lúpica (NL) y artritis idiopática juvenil (AIJ) frente a una molécula construida a partir de epítopes T de alérgenos de A. lumbricoides. MÉTODOS: Se diseñó y expresó una proteína multi-epítopes sintética (MP1), a partir de alérgenos de A. lumbricoides y ácaros del polvo doméstico. Mediante ELISA indirecto, se evaluaron las reactividades IgE anti-MP1 y al extracto de cuerpo entero de Ascaris lumbricoides, en sueros de pacientes con nefritis lúpica (NL; n=25), diabetes tipo 1 (T1D; n=10) y artritis idiopática juvenil (AIJ; n=10), procedentes del Caribe colombiano. Se excluyeron los individuos con poliautoinmunidad. Todos los pacientes fueron remitidos al estudio por su médico especialista. RESULTADOS: La IgE frente al extracto de cuerpo completo de A. lumbricoides mostró concentraciones de 484,2 ng/ml (RIQ: 203,4) en pacientes con AIJ; 325,6 ng/ml (RIQ: 179,3) en individuos con NL; y 424,7 ng/ml (RIQ: 80,1) en el grupo con DT1. Por otra parte, la reactividad de IgE anti-MP1 fue de 126,4 ng/ml (RIQ: 90,9) en los pacientes con AIJ; 130,7 ng/ml (RIQ: 94,8) en los individuos con NL; y 148,8 ng/ml (RIQ: 102,1) en el grupo con DT1. Aunque no se observaron diferencias estadísticas entre los grupos de pacientes, la reactividad IgE anti- MP1 en todos los pacientes (n: 45) (mediana de IgE: 134,2 ng/ml; RIQ: 100), fue significativamente inferior en comparación con el extracto de Ascaris (mediana de IgE: 380,7 ng/ml; RIQ: 175,8); (W: 0,732; p-valor: 1,034x10-7). CONCLUSIONES: Estos resultados preliminares sugieren que MP1 mostró propiedades antigénicas con baja reactividad IgE, en comparación con el extracto de Ascaris lumbricoides en individuos con enfermedades autoinmunes. Se necesitan más estudios para comprender mejor la respuesta inmunitaria inducida por esta molécula.
Assuntos
Alérgenos , Ascaris lumbricoides , Imunoglobulina E , Humanos , Animais , Ascaris lumbricoides/imunologia , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Alérgenos/imunologia , Feminino , Masculino , Doenças Autoimunes/imunologia , Doenças Autoimunes/sangue , Adolescente , Criança , Epitopos de Linfócito T/imunologia , AdultoRESUMO
OBJECTIVE: To quantify the production of Th1/Th2/Th17 cytokines induced by Ascaris lumbricoides antigens in peripheral blood mononuclear cells (PBMCs) using a multiplex technique. METHODS: PBMCs were cultured from individuals with mild A. lumbricoides infection (n = 20) and uninfected individuals (n = 21) and stimulated with A. lumbricoides extract (ExtAscaris), a mix of anti-CD2/CD3/CD28 (CDmix) as a positive control, and only medium (negative control). Cytokines in the supernatants were measured using the BD™ Cytometric Bead Array Human Th1/Th2/Th17 kit, to identify IFN-γ, TNF, IL-10, IL-6, IL-4, IL-2, and IL-17A. Readings were taken on a spectral cytometer (Northern Lights, Cytek, USA), and analysis was performed using R software with packages "tidyverse," "beadplexr," "flowCore," and "arsenal." Cytokine concentrations were calculated using a 5-parameter logistic curve. The t-test was used to compare cases and controls, and statistical significance was set at p < 0.05. The study was approved by the Ethics Committee of the University of Cartagena and the participants provided informed consent. This study was financially supported by the Colombian Sistema General de Regalías under the BPIN2020000100405 - BPIN2020000100364. RESULTS: Efficient fluorescence intensity extraction for each cytokine was achieved using detection channel R8 and the "mclust" clustering model (Figure 1). No significant differences were found in the levels of the seven cytokines between cases and controls (Figure 2). Although the IFN-γ response to ExtAscaris was higher in cases than in controls (252.5 ng/mL vs. 173.1 ng/mL), the difference was not significant. IL-17A (detection limit: 18.9 pg/mL) was more detectable in cases than controls (5 cases, 23% vs. 2 controls, 9.5%). IL-4 was only detected in the supernatants from CDmix-stimulated cultures but not with the Ascaris extract (Figure 2). CONCLUSIONS: The multiplex technique using spectral flow cytometry combined with open-source software analysis proved applicable for quantifying cytokines induced by A. lumbricoides antigens in PBMCs. However, a more sensitive method is needed to evaluate IL-4 response in the context of ascariasis. The results did not reveal significant differences in cytokine production between cases and controls for the evaluated stimuli.
OBJETIVOS: Cuantificar la producción de citoquinas Th1/Th2/Th17, inducida por antígenos de Ascaris lumbricoides en PBMCs, utilizando una técnica de multiplex. MÉTODOS: Se realizaron cultivos de PBMCs de personas con infección leve por A. lumbricoides (n = 20), y no infectadas (n = 21), y se estimularon con extracto de A. lumbricoides (ExtAscaris), un mix de anti-CD2/CD3/CD28 (CDmix), como control positivo, y solo medio (control negativo). Las citoquinas en los sobrenadantes, se midieron usando el estuche BD™ Cytometric Bead Array Human Th1/Th2/Th17, para identificar IFN-γ, TNF, IL-10, IL-6, IL-4, IL-2 e IL-17A. La lectura se realizó en un citómetro espectral (Northern Lights, Cytek, USA), y el análisis en software R, usando los paquetes tidyverse, beadplexr, flowCore y arsenal. Se calculó la concentración de citoquinas mediante ajuste de curva logística de cinco parámetros. Se empleó la prueba t para comparar casos y controles y una p < 0,05, se consideró como significativa. Se contó con autorización del Comité de Ética de la Universidad de Cartagena para hacer la investigación y con el consentimiento informado por parte de los participantes. Este trabajo fue financiado por el Sistema General de Regalías de Colombia, bajo el BPIN2020000100405 - BPIN2020000100364. RESULTADOS: Al utilizar el canal de detección R8 para identificar las citoquinas y el modelo de agrupamiento mclust, se extrajo eficientemente la intensidad de fluorescencia para cada citoquina (Figura 1). No se encontraron diferencias significativas en los niveles de las siete citoquinas entre casos y controles (Figura 2). Aunque la respuesta de IFN-, γ hacia ExtAscaris fue más alta en los casos de controles (252,5 ng/mL vs 173,1 ng/mL), la diferencia no fue significativa. La IL-17A (límite de detección: 18,9 pg/mL) fue más detectable en casos que en controles (cinco casos, 23% vs dos controles, 9,5%). La IL-4 solo se detectó en los sobrenadantes de cultivos estimulados con CDmix, pero no con el extracto de Ascaris (Figura 2). CONCLUSIONES: La técnica multiplex por citometría espectral, combinada con el análisis en software de licencia libre, se mostró aplicable para cuantificar citoquinas inducidas por antígenos de A. lumbricoides en PBMCs. Sin embargo, se requiere de un método más sensible para evaluar la respuesta de IL-4 en el contexto de la ascariasis. Los resultados no revelaron diferencias significativas en la producción de citoquinas entre casos y controles para los estímulos evaluados.
Assuntos
Ascaríase , Citocinas , Citometria de Fluxo , Humanos , Citocinas/sangue , Citometria de Fluxo/métodos , Ascaríase/imunologia , Ascaríase/diagnóstico , Masculino , Feminino , Adulto , Animais , Leucócitos Mononucleares/imunologia , Ascaris lumbricoides/imunologia , Adulto Jovem , Pessoa de Meia-Idade , Adolescente , Antígenos de Helmintos/imunologiaRESUMO
Genetic and epigenetic factors are considered to be critical for host-parasite interactions. There are limited data on the role of such factors during human infections with Ascaris lumbricoides. Here, we describe the potential role of genetic factors as determinants of the Th2 immune response to A. lumbricoides in Brazilian children. Stool samples were collected from the children to detect A. lumbricoides by microscopy and peripheral blood leukocytes (PBLs) were cultured in whole blood cultures for detection of cytokines (IL-5, IL-10, and IL-13) in vitro. Levels of anti-A. lumbricoides IgE and IgG4 were measured in plasma. DNA was extracted from PBLs and genotyped using Illumina 2.5 Human Omni Beadchip. Candidate genes associated with A. lumbricoides responses were identified and SNVs in these selected genes associated with the Th2 immune response to A. lumbricoides. Haplotype, gene expression, and epigenetic analyses were done to identify potential associations with Th2 immune responses. GWAS on samples from 1,189 children identified WSB1 as a candidate gene, and IL-21R was selected as a biologically relevant linked gene for further analysis. Variants in WSB1 and IL21R were associated with markers of Th2 immune responses: increased A. lumbricoides-specific IgE and IL-5/IL-13 by PBLs from infected compared to uninfected individuals. In infected children, WSB1 but not IL21R gene expression was suppressed and increased methylation was observed in the WSB1 promoter region. This is the first study to show an association between genetic variants in WSB1 and IL21R and Th2 immune responses during A. lumbricoides infections in children. WSB1/IL21R pathways could provide a potential target for the treatment of Th2-mediated diseases.
Assuntos
Ascaríase/imunologia , Ascaris lumbricoides/imunologia , Peptídeos e Proteínas de Sinalização Intracelular/genética , Receptores de Interleucina-21/genética , Células Th2/imunologia , Animais , Brasil , Células Cultivadas , Criança , Citocinas/metabolismo , Metilação de DNA , Feminino , Variação Genética , Estudo de Associação Genômica Ampla , Genótipo , Humanos , Imunidade Celular , Masculino , Regiões Promotoras Genéticas/genéticaRESUMO
To analyze the impact of Ascaris lumbricoides infection on the pathogenesis and diagnosis of allergic diseases, new allergens should be identified. We report the identification of a new Ascaris lumbricoides allergen, Asc l 5. The aim of this study was to evaluate the physicochemical and immunological features of the Asc l 5 allergen. We constructed an A. lumbricoides cDNA library and Asc l 5 was identified by immunoscreening. After purification, rAsc l 5 was physicochemically characterized. Evaluation of its allergenic activity included determination of Immunoglobulin E (IgE) binding frequency (in two populations: 254 children and 298 all-age subjects), CD203c based-basophil activation tests (BAT) and a passive cutaneous anaphylaxis (PCA) mouse model. We found by amino acid sequence analysis that Asc l 5 belongs to the SXP/RAL-2 protein family of nematodes. rAsc l 5 is a monomeric protein with an alpha-helical folding. IgE sensitization to rAsc l 5 was around 52% in general population; positive BAT rate was 60%. rAsc l 5 induced specific IgE production in mice and a positive PCA reaction. These results show that Asc l 5 has structural and immunological characteristics to be considered as a new allergen from A. lumbricoides.
Assuntos
Alérgenos/imunologia , Anticorpos Anti-Helmínticos/imunologia , Antígenos de Helmintos/imunologia , Ascaris lumbricoides/imunologia , Asma/imunologia , Hipersensibilidade/imunologia , Imunoglobulina E/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Asma/sangue , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Imunoglobulina E/sangue , Lactente , Camundongos , Camundongos Endogâmicos BALB C , Pessoa de Meia-Idade , Estudos Prospectivos , Adulto JovemRESUMO
Severe helminth infections are negatively associated to allergic diseases like asthma; therefore, the immunomodulatory properties of parasite-derived components have been analyzed, raising the possibility of their use as anti-inflammatory molecules. We evaluated the immunomodulatory properties of Ascaris lumbricoides recombinant cysteine protease inhibitor (rAl-CPI) in a mouse model of allergic airway inflammation induced by the house dust mite (HDM) Blomia tropicalis and its effects on human monocyte-derived dendritic cells (HmoDCs). The B. tropicalis sensitized/challenged mice developed extensive cellular airway inflammatory response, which was significantly reduced upon treatment with rAl-CPI prior to B. tropicalis sensitization, affecting particularly the perivascular/peribronchial infiltrate cells, eosinophils/neutrophils, and goblet cells. A significant decrease of Th2 cytokines, total, and specific IgE antibodies was observed in rAl-CPI treated mice. The antibody response was biased to IgG, mainly IgG2a. Administration of rAl-CPI-alone and rAl-CPI before mite sensitization were associated with a significant increase of regulatory T cells (Tregs) in spleen and elevated IL-10 levels in BAL and splenocytes culture supernatants, which was partially affected by anti-IL10 receptor use. In vitro, rAl-CPI showed a modulatory effect on HmoDCs, lowering the expression of HLA-DR, CD83, and CD86, while inducing IL-10 and IL-6 production. This suggests an inhibition of HmoDC maturation and a possible link with the inhibition of the allergic response observed in the murine model.
Assuntos
Ascaris lumbricoides/imunologia , Cistatinas/imunologia , Hipersensibilidade/imunologia , Inflamação/imunologia , Alérgenos/imunologia , Animais , Asma/imunologia , Citocinas/imunologia , Células Dendríticas/imunologia , Modelos Animais de Doenças , Eosinófilos/imunologia , Feminino , Humanos , Imunoglobulina G/imunologia , Interleucina-10/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Neutrófilos/imunologia , Pyroglyphidae/imunologia , Linfócitos T Reguladores/imunologiaRESUMO
Host-parasite interactions in diabetic patients might influence diabetes complications and intestinal parasitosis. The aim was to investigate the occurrence of enteroparasites in individuals with diabetes types 1 and 2. A descriptive study was designed to estimate frequencies of parasites and to compare them in individuals with diabetes types 1 and 2 from two Health Centers and one hospital in the Federal District of Brazil. Patients were allocated to the study by convenience. Three fecal samples of 156 diabetic individuals (120 type 1 and 36 type 2) were analyzed using two parasitological methods. Enteroparasites or commensals frequency in diabetics was 64%. Diabetics infected with up to six species of intestinal parasites or commensals were found. Frequencies of Ascaris lumbricoides and Giardia lamblia were higher in individuals with type 2 diabetes. The lower frequency of A. lumbricoides found in type 1 diabetes may be related to a strong Th2 response to parasites. Autoimmune response developed in type 1 diabetic individuals characterized by the production of Th1 cytokines could explain low frequency of G. lamblia. High frequency of parasites found in type 2 diabetes emphasizes the importance of periodic parasitological examinations in these individuals.
Assuntos
Ascaríase/epidemiologia , Ascaris lumbricoides/isolamento & purificação , Diabetes Mellitus Tipo 1/epidemiologia , Diabetes Mellitus Tipo 2/epidemiologia , Giardia lamblia/isolamento & purificação , Giardíase/epidemiologia , Interações Hospedeiro-Parasita/fisiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Ascaríase/complicações , Ascaris lumbricoides/imunologia , Brasil/epidemiologia , Criança , Pré-Escolar , Diabetes Mellitus Tipo 1/complicações , Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/parasitologia , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/imunologia , Diabetes Mellitus Tipo 2/parasitologia , Feminino , Giardia lamblia/imunologia , Giardíase/complicações , Interações Hospedeiro-Parasita/imunologia , Humanos , Lactente , Enteropatias Parasitárias/epidemiologia , Enteropatias Parasitárias/imunologia , Enteropatias Parasitárias/parasitologia , Masculino , Pessoa de Meia-Idade , Prevalência , Adulto JovemRESUMO
The aim of the present study was to evaluate associations between intestinal parasitic infection with intestinal and systemic inflammatory markers in school-aged children with high rates of obesity. Plasma concentrations of C-Reactive Protein (CRP), leptin, TNF-α, IL-6 and IL-10 were measured as systemic inflammation markers and count of stool leukocytes as marker of intestinal inflammation in 291 children (6-10 years). Intestinal parasitic infection was measured by stool examination. Logistic regression analyses were performed to determine the odds of having high inflammatory markers for each parasite or group of parasites as compared to parasite-free children while adjusting for sex, age, mother's educational level and percentage of body fat. The prevalence of soil-transmitted helminths and intestinal protozoa infections was 12% and 36%, respectively. Parasitic infection was not associated with CRP, IL-6, IL-10 or TNF-α. Children infected with Ascaris lumbricoides (aOR: 5.91, 95% CI: 1.97-17.70) and Entamoeba coli (aOR: 8.46, 95% CI: 2.85-25.14) were more likely to have higher stool leucocytes than parasite-free children. Children with multiple infections (aOR: 10.60, 95% CI: 2.85-25.14) were more likely to have higher leptin concentrations than parasite-free children. Intestinal parasitic infection was not associated with systemic inflammation, but was associated with intestinal inflammation. Having multiple infections were associated with higher leptin concentrations.
Assuntos
Helmintíase/sangue , Helmintíase/imunologia , Enteropatias Parasitárias/sangue , Enteropatias Parasitárias/imunologia , Intestinos/parasitologia , Leptina/sangue , Animais , Ascaris lumbricoides/imunologia , Proteína C-Reativa/metabolismo , Criança , Estudos Transversais , Entamoeba histolytica/imunologia , Fezes/parasitologia , Feminino , Helmintíase/parasitologia , Humanos , Inflamação , Interleucina-10/sangue , Interleucina-6/sangue , Enteropatias Parasitárias/parasitologia , Masculino , Obesidade/complicações , Prevalência , Inquéritos e Questionários , Fator de Necrose Tumoral alfa/sangueRESUMO
BACKGROUND: The dissociation between specific IgE and skin prick test reactivity to aeroallergens, a common finding in populations living in low and middle-income countries, has important implications for the diagnosis and treatment of allergic diseases. Few studies have investigated the determinants of this dissociation. In the present study, we explored potential factors explaining this dissociation in children living in an urban area of Northeast Brazil, focusing in particular on factors associated with poor hygiene. METHODS: Of 1445 children from low income communities, investigated for risk factors of allergies, we studied 481 with specific IgE antibodies to any of Blomia tropicalis, Dermatophagoides pteronyssinus, Periplaneta americana and Blatella germanica allergens. Data on demographic, environmental and social exposures were collected by questionnaire; serum IgG and stool examinations were done to detect current or past infections with viral, bacterial, protozoan and intestinal helminth pathogens. We measured atopy by skin prick testing (SPT) and specific IgE (sIgE) to aerollergens in serum (by ImmunoCAP). SIgE reactivity to B. tropicalis extract depleted of carbohydrates was measured by an in-house ELISA. Total IgE was measured by in house capture ELISA. SNPs were typed using Illumina Omni 2.5. RESULTS: Negative skin prick tests in the presence of specific IgE antibodies were frequent. Factors independently associated with a reduced frequency of positive skin prick tests were large number of siblings, the presence of IgG to herpes simplex virus, Ascaris lumbricoides and Trichuris trichiura infections, living in neighborhoods with infrequent garbage collection, presence of rodents and cats in the household and sIgE reactivity to glycosylated B. tropicalis allergens. Also, SNP on IGHE (rs61737468) was negatively associated with SPT reactivity. CONCLUSIONS: A variety of factors were found to be associated with decreased frequency of SPT such as unhygienic living conditions, infections, total IgE, IgE response to glycosylated allergens and genetic polymorphisms, indicating that multiple mechanisms may be involved. Our data, showing that exposures to an unhygienic environment and childhood infections modulate immediate allergen skin test reactivity, provide support for the "hygiene hypothesis".
Assuntos
Alérgenos/imunologia , Hipersensibilidade/imunologia , Imunoglobulina E/imunologia , Testes Cutâneos/métodos , Animais , Ascaris lumbricoides/imunologia , Brasil , Gatos , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Fezes/microbiologia , Fezes/parasitologia , Fezes/virologia , Humanos , Hipersensibilidade/sangue , Hipersensibilidade/diagnóstico , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Cadeias épsilon de Imunoglobulina/genética , Cadeias épsilon de Imunoglobulina/imunologia , Polimorfismo de Nucleotídeo Único/imunologia , Receptores de IgE/genética , Receptores de IgE/imunologia , Roedores , Simplexvirus/imunologia , Trichuris/imunologia , Saúde da População Urbana/estatística & dados numéricosRESUMO
BACKGROUND: Asthma is a common disease in the world and vitamin D (Vit-D) has been associated with the presence and severity of this disease. OBJECTIVE: To establish the association between levels of Vit-D and IgE response in schoolchildren with asthma living in four cities in Colombia. METHODS: Case-control study in 1340 schoolchildren (687 asthmatic and 653 controls) from communities in extreme poverty in Barranquilla, Cartagena, Santa Marta, and Montería. Serum concentrations of Vit-D, total IgE, and anti-Dermatophagoides farinae, Periplaneta americana, and Ascaris lumbricoides (AL) specific IgE were measured. RESULTS: Controls reported higher concentrations of Vit-D [61.9 ± 28.4 ng/mL] than cases [53 ± 23.3 ng / mL] (p < 0.05). Total IgE was higher in cases (p < 0.05). Only anti-AL IgE showed a clear difference: in controls, optical density was 0.27 ± 0.25; in cases, 0.22 ± 0.24 (p < 0.05). Vit-D showed differences between cases and controls in each population. CONCLUSIONS: An association could not be demonstrated between Vit-D deficiency and asthma, as total IgE was elevated in patients and controls. The results suggest that Vit-D influences the specif IgE response in poor asthmatic children in areas endemic for helminthiasis.
Antecedentes: El asma es una enfermedad frecuente en el mundo y la vitamina D (Vit-D) se ha asociado con la presencia y severidad de esta enfermedad. Objetivo: Establecer la asociación entre los niveles de Vit-D y la respuesta IgE en escolares con asma residentes de cuatro ciudades colombiananas. Métodos: Estudio de casos y controles en 1340 escolares (687 asmáticos y 653 controles) de comunidades en extrema pobreza de Barranquilla, Cartagena, Santa Marta y Montería. Se midieron las concentraciones séricas de Vit-D, IgE total e IgE específica anti Dermatofagoides farinae, Periplaneta americana y Ascaris lumbricoides (AL). Resultados: Los controles reportaron concentraciones mayores de Vit-D [61.9 ± 28.4 ng/mL] que los casos [53 ± 23.3 ng/mL] (p<0.05). La IgE total fue mayor en los casos (p<0.05). Solo IgE anti-AL mostró una diferencia clara: controles, densidad óptica 0.27 ± 0.25; casos 0.22 ± 0.24 (p<0.05). La Vit-D presentó diferencias entre casos y controles en cada población. Conclusiones: No se pudo demostrar la asociación entre deficiencia de Vit-D y asma, dado que la IgE total estuvo elevada en los pacientes y en los controles. Los resultados sugieren que la Vit-D influye en la respuesta IgE específica en niños asmáticos pobres en zonas endémicas para helmintiasis.
Assuntos
Asma/sangue , Asma/imunologia , Imunoglobulina E/imunologia , Áreas de Pobreza , Deficiência de Vitamina D/imunologia , Vitamina D/sangue , Alérgenos , Animais , Ascaris lumbricoides/imunologia , Estudos de Casos e Controles , Criança , Colômbia , Dermatophagoides farinae/imunologia , Humanos , Imunoglobulina E/sangue , Periplaneta/imunologiaRESUMO
Infection with intestinal helminths results in immunological changes that influence co-infections, and might influence fecundity by inducing immunological states affecting conception and pregnancy. We investigated associations between intestinal helminths and fertility in women, using 9 years of longitudinal data from 986 Bolivian forager-horticulturalists, experiencing natural fertility and 70% helminth prevalence. We found that different species of helminth are associated with contrasting effects on fecundity. Infection with roundworm (Ascaris lumbricoides) is associated with earlier first births and shortened interbirth intervals, whereas infection with hookworm is associated with delayed first pregnancy and extended interbirth intervals. Thus, helminths may have important effects on human fertility that reflect physiological and immunological consequences of infection.
Assuntos
Fertilidade/imunologia , Número de Gestações/imunologia , Helmintíase/imunologia , Enteropatias Parasitárias/imunologia , Fatores Etários , Animais , Ascaríase/epidemiologia , Ascaríase/imunologia , Ascaris lumbricoides/imunologia , Bolívia/epidemiologia , Coinfecção , Feminino , Fertilidade/fisiologia , Número de Gestações/fisiologia , Humanos , Enteropatias Parasitárias/epidemiologia , Gravidez , Prevalência , Adulto JovemAssuntos
Antígenos de Helmintos/administração & dosagem , Extratos Celulares/administração & dosagem , Eosinófilos/imunologia , Mucosa Respiratória/patologia , Rinite Alérgica/terapia , Angiostrongylus/imunologia , Animais , Ascaris lumbricoides/imunologia , Contagem de Células , Extratos Celulares/imunologia , Movimento Celular , Células Cultivadas , Modelos Animais de Doenças , Feminino , Humanos , Imunização , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Rinite Alérgica/imunologiaRESUMO
BACKGROUND: Helminth infections and allergies are diseases with intense Th2 lymphocytes participation and characterised by a high IgE and Interleukin-(IL) IL-4, IL-5 production and eosinophilia. However, helminths also induce IL-10 production, which may alter the outcome of allergic diseases in infected patients. OBJECTIVE: This experimental study analyses the relationship between IL-10 production by cell culture from geohelminth infected and non-infected children and specific IgE to Ascaris lumbricoides (Asc) or Blomia tropicalis (BT). METHODS: IL-10 content in supernatant from peripheral blood mononuclear cell culture from nine helminth infected and eleven non-infected patients was determined by ELISA after in vitro stimulation with Asc or BT extracts. RESULTS: A positive association was observed between total IgE levels and anti-Ascaris and anti-Blomia tropicalis specific IgE, independent of infection status. For both helminth-infected and non-infected groups, there was no difference in IL-10 production in response to Asc extract, even though anti-Ascaris IgE levels were higher in the latter group. In response to BT stimulus, a lower production of IL-10 by the geohelminth-infected group was observed, but with no relationship between IL-10 production and specific IgE to BT. CONCLUSION: The results suggest that anti-Ascaris IgE in non-infected patients may be associated to a resistance to parasites. Levels of specific IgE to parasite antigens or B. tropicalis allergen were not impaired by IL-10 production in children from an urban area in which geohelminthiasis is endemic.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Ascaríase/imunologia , Ascaris lumbricoides/imunologia , Hipersensibilidade/imunologia , Imunoglobulina E/sangue , Interleucina-10/sangue , Leucócitos Mononucleares/imunologia , Acaridae/imunologia , Alérgenos/imunologia , Animais , Antígenos de Helmintos/imunologia , Células Cultivadas , Criança , Feminino , Humanos , Proteínas de Insetos/imunologia , Leucócitos Mononucleares/parasitologia , Masculino , População , Testes Cutâneos , População UrbanaRESUMO
Helminth infections and allergy have evolutionary and clinical links. Infection with the nematode Ascaris lumbricoides induces IgE against several molecules including invertebrate pan-allergens. These antibodies influence the pathogenesis and diagnosis of allergy; therefore, studying parasitic and non-parasitic allergens is essential to understand both helminth immunity and allergy. Glutathione transferases (GSTs) from cockroach and house dust mites are clinically relevant allergens and comparative studies between them and the GST from A. lumbricoides (GSTA) are necessary to evaluate their allergenicity. We sought to analyze the allergenic potential of GSTA in connection with the IgE response to non-parasitic GSTs. IgE to purified GSTs from Ascaris (nGSTA and rGSTA), house dust mites (rDer p 8, nBlo t 8 and rBlo t 8), and cockroach (rBla g 5) was measured by ELISA in subjects from Cartagena, Colombia. Also, multidimensional proteomic approaches were used to study the extract of A. lumbricoides and investigate the existence of GST isoforms. We found that among asthmatics, the strength of IgE levels to GSTA was significantly higher than to mite and cockroach GSTs, and there was a strong positive correlation between IgE levels to these molecules. Specific IgE to GSTA was found in 13.2% of controls and 19.5% of asthmatics. In addition nGSTA induced wheal and flare in skin of sensitized asthmatics indicating that it might be of clinical relevance for some patients. Frequency and IgE levels to GSTA were higher in childhood and declined with age. At least six GST isoforms in A. lumbricoides bind human IgE. Four isoforms were the most abundant and several amino acid substitutions were found, mainly on the N-terminal domain. In conclusion, a new allergenic component of Ascaris has been discovered; it could have clinical impact in allergic patients and influence the diagnosis of mite and cockroach allergy in tropical environments.
Assuntos
Ascaris lumbricoides/imunologia , Asma/imunologia , Baratas/imunologia , Glutationa Transferase/imunologia , Proteínas de Helminto/imunologia , Imunoglobulina E/imunologia , Pyroglyphidae/imunologia , Adolescente , Adulto , Alérgenos/química , Alérgenos/genética , Alérgenos/imunologia , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Reações Antígeno-Anticorpo , Antígenos de Dermatophagoides/química , Antígenos de Dermatophagoides/genética , Antígenos de Dermatophagoides/imunologia , Ascaris lumbricoides/química , Ascaris lumbricoides/enzimologia , Asma/genética , Baratas/química , Colômbia , Feminino , Glutationa Transferase/química , Glutationa Transferase/genética , Proteínas de Helminto/química , Proteínas de Helminto/genética , Humanos , Imunoquímica , Proteínas de Insetos/química , Proteínas de Insetos/genética , Proteínas de Insetos/imunologia , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/imunologia , Masculino , Pessoa de Meia-Idade , Modelos Moleculares , Dados de Sequência Molecular , Estrutura Terciária de Proteína , Proteômica , Pyroglyphidae/químicaRESUMO
INTRODUCTION: Ascaris lumbricoides-infected patients present lower prevalence of severe atopic dermatitis. METHODS: Peripheral blood of infected children with atopic dermatitis was assessed by flow cytometry of the frequency of Th1 and Th2 cells through the expression of CXCR3 and CCR4 chemokine receptors, respectively. RESULTS: Helminth-free patients with atopic dermatitis presented a high frequency of CCR4+Th2 cells. Parasitized patients with atopic dermatitis showed a lower frequency of CXCR3+Th1 cells compared to infected individuals only. CONCLUSIONS: Ascariasis modifies the blood traffic of Th2 cells in atopic dermatitis patients, while the allergic disease down-regulates the traffic of Th1 cells in parasitized patients.
INTRODUÇÃO: Pacientes infectados com Ascaris lumbricoides apresentam menor prevalência de dermatite atópica grave. MÉTODOS: Sangue periférico de crianças infectadas com dermatite atópica foi analisado por citometria de fluxo quanto à frequência de células Th1 e Th2 pela expressão de receptores de quimiocina CXCR3 e CCR4, respectivamente. RESULTADOS: Pacientes sem helmintos com dermatite atópica apresentaram alta frequência de células Th2CCR4+. Pacientes parasitados com dermatite atópica apresentaram menor frequência de células Th1CXCR3+ comparados aos indivíduos apenas infectados. CONCLUSÕES: Ascaridiases altera o tráfego sanguíneo de células Th2 em pacientes com dermatite atópica, enquanto a doença alérgica diminui o tráfego de células Th1 em pacientes parasitados.
Assuntos
Adolescente , Animais , Criança , Pré-Escolar , Humanos , Ascaríase/imunologia , Ascaris lumbricoides/imunologia , Dermatite Atópica/imunologia , /imunologia , /imunologia , Células Th1/imunologia , /imunologia , Ascaríase/complicações , Estudos de Casos e Controles , Dermatite Atópica/complicações , Citometria de Fluxo , Fezes/parasitologia , Índice de Gravidade de DoençaRESUMO
La poliaglutinabilidad de los glóbulos rojos puede deberse al desenmascaramiento del antígeno T críptico debido a la acción de neuraminidasas microbianas que eliminan residuos terminales de ácido siálico en la membrana del hematíe. En experiencias preliminares se demostró que Ascaris lumbricoides capta ácido siálico del eritrocito y que las suspensiones globulares en medio enzimático, incubadas con este parásito, pierden totalmente la capacidad de agregación. El objetivo fue estudiar la exposición del antígeno T eritrocitario, por acción de A. lumbricoides sobre la carga aniónica de glóbulos rojos deficientes en ácido siálico. Se trabajó con 48 extractos de ejemplares adultos del parásito ([EA]) y un concentrado de larvas L1/ L2 ([CLAL]):1300-1500 larvas/mL). Se utilizaron eritrocitos Grupo O en medio enzimático de bromelina (GRB) y eritrocitos Control en medio salino (GRC). El sedimento globular de GRB se incubó con igual volumen de [EA]/ [CLAL] y el de GRC con solución fisiológica durante 120 minutos a 37 ºC. Se realizaron Pruebas de Aglutinación en Placa y en Tubo, enfrentando los GRB y GRC a sueros humanos de adulto y de cordón. Los resultados mostraron que el 33,33% de los GRB incubados con [EA] y el 66,67% de los GRB incubados con [CLAL] aglutinaron con suero de adulto, pero no con suero de cordón. Los GRC no aglutinaron con ninguno de los sueros. Es la primera vez que se comunica la exposición del antígeno T eritrocitario por acción de un parásito. La activación T podría producir autoaglutinación y hemólisis en el hombre adulto y representaría un factor de riesgo transfusional en la población infantil.
Red cell polyagglutination may be due to the unmasking of the cryptic T antigen by the action of microbial neuraminidases, which remove terminal sialic acid residues in the erythrocyte membrane. Previous experiences showed that Ascaris lumbricoides capture erythrocyte sialic acid and thatglobular suspensions in enzymatic medium, incubated with this parasite, completely lost the ability to aggregate. The aim of this work was to study the erythrocyte T antigen exposure by A. lumbricoides action on the anionic charge of sialic acid deficient red cells. Studies were done on 48 adult specimen parasite extracts ([AE]) and an L1/ L2 larvae concentrate ([ALLC]: 1300-1500 larvae/mL). Group O red cells in bromelain enzymatic medium (RCB) and Control erythrocytes in saline medium (RCC) were used. The RCB sediment was incubated with an equal volume of [AE]/ [ALLC] and the RCC sediment with physiological solution during 120 minutes at 37 ºC. Plate and Tube Agglutination Tests were performed, contrasting RCB and RCC with adult and cord human sera. The results showed that 33.33% of the RCB incubated with [AE] and 66.67% of the RCB incubated with [LCAL] agglutinated with serum from adult, but not cord serum. RCB were not agglutinated with any of the sera. It is the first time that erythrocyte T antigen exposure by a parasite action is communicated. T activation could produce autoagglutination and haemolysis in the adult and would represent a transfusion risk factor in the child population.
A poliaglutinabilidade dos glóbulos vermelhos pode ser resultado do desmascaramento do antígeno críptico T devido à ação de neuraminidases microbianas que eliminam resíduos terminais de ácido siálico na membrana da hemácia. Em experiências preliminares foi demonstrado que Ascaris lumbricoides capta ácido siálico do eritrócito e que as suspensões globulares em meio enzimático, incubadas com esta parasita, perdem totalmente a capacidade de agregação. O objetivo foi estudar a exposição do antígeno T eritrocitário, por ação de A. lumbricoides sobre a carga aniônica de glóbulos vermelhos deficientes em ácido siálico. Trabalhou-se com 48 extratos de exemplares adultos da parasita ([EA]) e um concentrado de larvas L1/ L2 ([CLAL]):1300-1500 larvas/mL). Foram utilizados eritrócitos Grupo O em meio enzimático de bromelina (GVB) e eritrócitos Controle em meio salino (GVC). O sedimento globular de GVB foi incubado com igual volume de [EA]/ [CLAL] e o de GVC com solução fisiológica durante 120 minutos a 37 ºC. Foram realizados Testes de Aglutinação em Placa e em Tubo, enfrentando os GVB e GVC a soros humanos de adulto e de cordão. Os resultados mostraram que 33,33% dos GVB incubados com [EA] e 66,67% dos GVB incubados com [CLAL] aglutinaram com soro de adulto, mas não com soro de cordão. Os GVC não aglutinaram com nenhum dos soros. É a primeira vez que se comunica a exposição do antígeno T eritrocitário por ação de uma parasita. A ativação T poderia produzir autoaglutinação e hemólise no homem adulto e representaria um fator de risco transfusional na população infantil.
Assuntos
Humanos , Ascaríase , Ascaris lumbricoides/imunologia , Ascaris lumbricoides/fisiologia , Ascaris lumbricoides/parasitologia , Eritrócitos , Receptores de Antígenos de Linfócitos T/antagonistas & inibidores , Receptores de Antígenos de Linfócitos T/fisiologiaRESUMO
BACKGROUND: Children of mothers infected with soil-transmitted helminths (STH) may have an increased susceptibility to STH infection. METHODS AND FINDINGS: We did a case-control study nested in a birth cohort in Ecuador. Data from 1,004 children aged 7 months to 3 years were analyzed. Cases were defined as children with Ascaris lumbricoides and/or Trichuris trichiura, controls without. Exposure was defined as maternal infection with A. lumbricoides and/or T. trichiura, detected during the third trimester of pregnancy. The analysis was restricted to households with a documented infection to control for infection risk. Children of mothers with STH infections had a greater risk of infection compared to children of uninfected mothers (adjusted OR 2.61, 95% CI: 1.88-3.63, p<0.001). This effect was particularly strong in children of mothers with both STH infections (adjusted OR: 5.91, 95% CI: 3.55-9.81, p<0.001). Newborns of infected mothers had greater levels of plasma IL-10 than those of uninfected mothers (p=0.033), and there was evidence that cord blood IL-10 was increased among newborns who became infected later in childhood (p=0.060). CONCLUSION: Our data suggest that maternal STH infections increase susceptibility to infection during early childhood, an effect that was associated with elevated IL-10 in cord plasma.
Assuntos
Ascaríase/epidemiologia , Ascaris lumbricoides/isolamento & purificação , Suscetibilidade a Doenças , Complicações Parasitárias na Gravidez/epidemiologia , Tricuríase/epidemiologia , Trichuris/isolamento & purificação , Animais , Ascaríase/imunologia , Ascaris lumbricoides/imunologia , Estudos de Casos e Controles , Pré-Escolar , Estudos de Coortes , Equador/epidemiologia , Feminino , Humanos , Lactente , Interleucina-10/sangue , Masculino , Gravidez , Medição de Risco , Tricuríase/imunologia , Trichuris/imunologiaRESUMO
INTRODUCTION: Ascaris lumbricoides-infected patients present lower prevalence of severe atopic dermatitis. METHODS: Peripheral blood of infected children with atopic dermatitis was assessed by flow cytometry of the frequency of Th1 and Th2 cells through the expression of CXCR3 and CCR4 chemokine receptors, respectively. RESULTS: Helminth-free patients with atopic dermatitis presented a high frequency of CCR4+Th2 cells. Parasitized patients with atopic dermatitis showed a lower frequency of CXCR3+Th1 cells compared to infected individuals only. CONCLUSIONS: Ascariasis modifies the blood traffic of Th2 cells in atopic dermatitis patients, while the allergic disease down-regulates the traffic of Th1 cells in parasitized patients.
Assuntos
Ascaríase/imunologia , Ascaris lumbricoides/imunologia , Dermatite Atópica/imunologia , Receptores CCR4/imunologia , Receptores CXCR3/imunologia , Células Th1/imunologia , Células Th2/imunologia , Adolescente , Animais , Ascaríase/complicações , Estudos de Casos e Controles , Criança , Pré-Escolar , Dermatite Atópica/complicações , Fezes/parasitologia , Citometria de Fluxo , Humanos , Índice de Gravidade de DoençaRESUMO
BACKGROUND: Helminth co-infection in humans is common in tropical regions of the world where transmission of soil-transmitted helminths such as Ascaris lumbricoides, Trichuris trichiura, and the hookworms Necator americanus and Ancylostoma duodenale as well as other helminths such as Schistosoma mansoni often occur simultaneously. METHODOLOGY: We investigated whether co-infection with another helminth(s) altered the human immune response to crude antigen extracts from either different stages of N. americanus infection (infective third stage or adult) or different crude antigen extract preparations (adult somatic and adult excretory/secretory). Using these antigens, we compared the cellular and humoral immune responses of individuals mono-infected with hookworm (N. americanus) and individuals co-infected with hookworm and other helminth infections, namely co-infection with either A. lumbricoides, Schistosoma mansoni, or both. Immunological variables were compared between hookworm infection group (mono- versus co-infected) by bootstrap, and principal component analysis (PCA) was used as a data reduction method. CONCLUSIONS: Contrary to several animal studies of helminth co-infection, we found that co-infected individuals had a further downmodulated Th1 cytokine response (e.g., reduced INF-γ), accompanied by a significant increase in the hookworm-specific humoral immune response (e.g. higher levels of IgE or IgG4 to crude antigen extracts) compared with mono- infected individuals. Neither of these changes was associated with a reduction of hookworm infection intensity in helminth co-infected individuals. From the standpoint of hookworm vaccine development, these results are relevant; i.e., the specific immune response to hookworm vaccine antigens might be altered by infection with another helminth.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Ascaríase/imunologia , Coinfecção/imunologia , Linfócitos/imunologia , Necator americanus/imunologia , Necatoríase/imunologia , Esquistossomose mansoni/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Antígenos de Helmintos , Ascaríase/parasitologia , Ascaris lumbricoides/imunologia , Criança , Coinfecção/parasitologia , Feminino , Humanos , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Interferon gama/metabolismo , Masculino , Pessoa de Meia-Idade , Necatoríase/parasitologia , Schistosoma mansoni/imunologia , Esquistossomose mansoni/parasitologia , Adulto JovemRESUMO
BACKGROUND: Two conditions are used as markers of atopy: the presence of circulating anti-allergen IgE antibodies and the presence of positive skin prick test (SPT) reactions to allergenic extracts. The correlation between these conditions is not absolute. This study aimed at investigating immunological parameters that may mediate this lack of correlation. Individuals whose sera contained anti-B. tropicalis extract IgE antibodies (α-BtE IgE) were divided into two groups, according to the presence or absence of skin reactivity to B. tropicalis extract (BtE). The following parameters were investigated: total IgE levels; α-BtE IgE levels; an arbitrary α-BtE IgE/total IgE ratio; the proportion of carbohydrate-reactive α-BtE IgE; the proportion of α-BtE IgE that reacted with Ascaris lumbricoides extract (AlE); the production of IL-10 by BtE- and AlE-stimulated peripheral blood cells (PBMC). RESULTS: Total IgE levels were similar in the two groups, but α-BtE IgE was significantly higher in the SPT-positive group (SPT+). A large overlap of α-BtE IgE levels was found in individuals of both groups, indicating that these levels alone cannot account for the differences in SPT outcome. Individuals of the two groups did not differ, statistically, in the proportion of α-BtE IgE that reacted with carbohydrate and in the production of IL-10 by BtE- and AlE-stimulated PBMC. Both groups had part of α-BtE IgE activity absorbed out by AlE, indicating the existence of cross-reactive IgE antibodies. However, the α-BtE IgE from the SPT-negative individuals (SPT-) was more absorbed with AlE than the α-BtE IgE from the SPT+ individuals. This finding may be ascribed to avidity differences of the α-BtE IgE that is present in the two groups of individuals, and could occur if at least part of the α-BtE IgE from the SPT- individuals were elicited by A. lumbricoides infection. CONCLUSION: The present results suggest that a low ratio of specific IgE to total IgE levels (in a minority of individuals), and differences in α-BtE IgE avidities (which would have high affinities for A. lumbricoides antigens in SPT- than in SPT+ individuals) may play a role in the down-modulation of type-I hypersensitivity reaction against aeroallergens described in helminth-infected individuals.