RESUMO
Blomia tropicalis, Dermatophagoides pteronyssinus and D. farinae are prevalent house dust mites. Concanavalin A-binding components derived from B. tropicalis (Bt-ConA extract) are highly immunogenic in allergic diseases. The aim of the present study was to evaluate the humoral and cellular immune responses to B. tropicalis in mite-sensitized patients. A total of 137 patients with allergic rhinitis with/without asthma and 109 non-atopic subjects were selected and analyzed by the skin prick test, and for total serum IgE and specific IgE levels to both Bt-total and Bt-ConA extracts, their proliferative response and cytokine (IFN-gamma and IL-5) production by peripheral blood mononuclear cells (PBMC) stimulated with both extracts. Skin prick test showed that 70% of the patients were sensitized to Bt (Bt+) and similar levels of specific IgE to Bt-total and Bt-ConA extracts were demonstrable in Bt+ patients. Significant PBMC proliferation was observed in response to Bt-total extract in Bt+, but not in Bt- patients and non-atopic subjects (P < 0.001). Bt-ConA extract induced increased proliferative responses in all patient groups compared to medium alone (P < 0.05), but these responses were significantly decreased in the presence of the mannopyranoside ConA inhibitor (P < 0.05). Significant IFN-gamma production was observed after Bt-ConA stimulation of Bt+ patients (P < 0.05), while Bt-total extract had no effect. IL-5 production was consistently detected in Bt+ patients after allergen-specific stimulation or with no stimulus, indicating that PBMC from allergic patients are prone to produce Th2 profile cytokines, spontaneously or inductively by allergen restimulation. These data showed that ConA-binding components isolated from B. tropicalis may contain relevant antigens that are involved in both humoral and cellular immune responses. However, without an additional purification procedure to eliminate the residual contamination with ConA, its use in immunotherapeutic procedures cannot be recommended.
Assuntos
Alérgenos/administração & dosagem , Antígenos de Dermatophagoides/administração & dosagem , Concanavalina A/administração & dosagem , Mitógenos/administração & dosagem , Rinite Alérgica Perene/imunologia , Adulto , Alérgenos/imunologia , Animais , Antígenos de Dermatophagoides/imunologia , Antígenos de Plantas , Estudos de Casos e Controles , Proliferação de Células , Concanavalina A/imunologia , Dessensibilização Imunológica , Feminino , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Interferon gama/biossíntese , Interleucina-5/biossíntese , Leucócitos Mononucleares/imunologia , Masculino , Ácaros/imunologia , Mitógenos/imunologia , Rinite Alérgica Perene/sangueRESUMO
Blomia tropicalis, Dermatophagoides pteronyssinus and D. farinae are prevalent house dust mites. Concanavalin A-binding components derived from B. tropicalis (Bt-ConA extract) are highly immunogenic in allergic diseases. The aim of the present study was to evaluate the humoral and cellular immune responses to B. tropicalis in mite-sensitized patients. A total of 137 patients with allergic rhinitis with/without asthma and 109 non-atopic subjects were selected and analyzed by the skin prick test, and for total serum IgE and specific IgE levels to both Bt-total and Bt-ConA extracts, their proliferative response and cytokine (IFN-ã and IL-5) production by peripheral blood mononuclear cells (PBMC) stimulated with both extracts. Skin prick test showed that 70 percent of the patients were sensitized to Bt (Bt+) and similar levels of specific IgE to Bt-total and Bt-ConA extracts were demonstrable in Bt+ patients. Significant PBMC proliferation was observed in response to Bt-total extract in Bt+, but not in Bt- patients and non-atopic subjects (P < 0.001). Bt-ConA extract induced increased proliferative responses in all patient groups compared to medium alone (P < 0.05), but these responses were significantly decreased in the presence of the mannopyranoside ConA inhibitor (P < 0.05). Significant IFN-ã production was observed after Bt-ConA stimulation of Bt+ patients (P < 0.05), while Bt-total extract had no effect. IL-5 production was consistently detected in Bt+ patients after allergen-specific stimulation or with no stimulus, indicating that PBMC from allergic patients are prone to produce Th2 profile cytokines, spontaneously or inductively by allergen restimulation. These data showed that ConA-binding components isolated from B. tropicalis may contain relevant antigens that are involved in both humoral and cellular immune responses. However, without an additional purification procedure to eliminate the residual contamination with...
Assuntos
Adulto , Animais , Feminino , Humanos , Masculino , Alérgenos/administração & dosagem , Antígenos de Dermatophagoides/administração & dosagem , Concanavalina A/administração & dosagem , Mitógenos/administração & dosagem , Rinite Alérgica Perene/imunologia , Alérgenos/imunologia , Antígenos de Dermatophagoides/imunologia , Estudos de Casos e Controles , Proliferação de Células , Concanavalina A/imunologia , Dessensibilização Imunológica , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Interferon gama/biossíntese , /biossíntese , Leucócitos Mononucleares/imunologia , Ácaros/imunologia , Mitógenos/imunologia , Rinite Alérgica Perene/sangueRESUMO
OBJECTIVE: This study aimed to evaluate the clinical efficacy and antibody response changes after specific immunotherapy (SIT) using Dermatophagoides pteronyssinus (Dpt) allergens with or without bacterial extracts in Brazilian mite-atopic patients. METHODS: One-hundred patients with allergic rhinitis were selected for a randomized double-blind, placebo-controlled trial and distributed into 4 groups: Dpt (Dpt allergen extract), Dpt+MRB (Dpt allergen plus mixed respiratory bacterial extracts), MRB (MRB extract only) and placebo. Rhinitis symptom and medication scores; skin prick test (SPT) to Dpt extract; and serum immunoglobulin (Ig) E, IgG4, and IgG1 levels to Dpt, Der p 1, and Der p 2 allergens were evaluated before and after a year of treatment. RESULTS: After 1 year, the SPT response was reduced in the Dpt group (P=.03), whereas IgE levels to Der p 2 decreased only in the Dpt (P = .048) and Dpt+MRB (P = .005) groups. IgG4 and IgG1 levels to Dpt and Der p 1 increased in the Dpt group (P < .05), whereas in the Dpt + MRB group the IgG1 level only increased to Dpt (P=.001) and the IgG4 only increased to Der p 1 (P=.049). IgE levels to Dpt decreased only in the MRB (P= .005) and Dpt + MRB (P= .001) groups. Rhinitis symptom and medication scores fell in all groups, including the placebo group (P<.001). CONCLUSIONS: SIT using Dpt extract alone was effective in reducing SPT response and IgE levels to Der p 2 allergen, while bacterial extracts induced decreases in IgE levels to whole Dpt extract. However, only groups receiving Dpt allergen had higher levels of IgG1 and IgG4 to Dpt and Der p 1 after a year of treatment.
Assuntos
Antígenos de Dermatophagoides/imunologia , Antígenos de Dermatophagoides/uso terapêutico , Bactérias/imunologia , Dermatophagoides pteronyssinus/imunologia , Imunoterapia , Rinite Alérgica Perene/terapia , Adolescente , Adulto , Animais , Bactérias/citologia , Brasil , Criança , Método Duplo-Cego , Combinação de Medicamentos , Feminino , Humanos , Imunoglobulinas/sangue , Imunoglobulinas/imunologia , Imunoterapia/efeitos adversos , Masculino , Pessoa de Meia-Idade , Rinite Alérgica Perene/sangue , Rinite Alérgica Perene/imunologiaRESUMO
UNLABELLED: Mite allergens are involved in airway sensitization and allergic diseases. We evaluated the exposure to house dust-mite (Der p 1 and Der f 1) and pet (Fel d 1 and Can f 1) allergens in hotels in Uberlândia, Midwestern Brazil. A total of 140 dust samples were collected from bedding (n = 98) and carpet (n = 42) of bedrooms in 20 hotels enrolled for this study. Geometric mean (GM) levels of Der f 1 (11.30 microg/g of dust; 95% CI: 8.34-15.30 microg/g) were significantly higher than Der p 1 (0.15 microg/g of dust; 95% CI: 0.13-0.18 microg/g) in bedding dust samples (P < 0.001), regardless of the hotel classes. Der f 1 levels were significantly higher in bedding (11.30 microg/g of dust; 95% CI: 8.34-15.30 microg/g) than carpet (6.32 microg/g of dust; 95% CI: 4.31-9.26 microg/g) dust samples (P < 0.05). High levels of Der f 1 (>10 microg/g of dust) were found in 58%, 76%, and 69% of dust samples from Simple, Economical, and Tourist/Superior hotels, respectively, while GM levels of Fel d 1 (0.11 microg/g of dust; 95% CI: 0.09-0.14 microg/g) and Can f 1 (0.30 microg/g of dust; 95% CI: 0.20-0.44 microg/g) were relatively low. These results indicate that Der f 1 is the predominant allergen in hotels in Uberlândia, especially in bedding dust samples, regardless of the hotel classes and could represent an important risk factor for exacerbation of allergic symptoms in previously mite-sensitized guests. PRACTICAL IMPLICATIONS: Mites and pets are important sources of indoor allergens. Most people spend the greatest part of their time indoors. Hotels can constitute an important allergen reservoir of the indoor environment and could represent an important risk for exacerbation of allergic symptoms in previously sensitized guests. Thus, hotels should also be included for planning indoor allergen avoidance as part of a global management strategy, especially in patients with respiratory allergy.
Assuntos
Poluição do Ar em Ambientes Fechados , Alérgenos/análise , Animais Domésticos , Hipersensibilidade , Ácaros/química , Viagem , Poluição do Ar em Ambientes Fechados/efeitos adversos , Poluição do Ar em Ambientes Fechados/análise , Alérgenos/imunologia , Animais , Roupas de Cama, Mesa e Banho , Brasil , Cidades , Exposição Ambiental , Pisos e Cobertura de Pisos , Habitação , Humanos , Hipersensibilidade/etiologia , Hipersensibilidade/imunologia , Hipersensibilidade/patologia , Fatores de RiscoRESUMO
Blomia tropicalis (Bt) and Dermatophagoides pteronyssinus (Dp) are the prevalent house dust mites in tropical countries and are associated with allergic diseases. Glycosylated antigens are highly immunogenic and involved in different pathologies. We evaluated the presence of IgE, IgG1, and IgG4 to concanavalin A-binding antigens (Bt-Con-A) isolated from Bt-total extract in sera of allergic and non-allergic subjects. Bt-total and Bt-Con-A extracts were evaluated by SDS-PAGE and ELISA for reacting with IgE, IgG1, and IgG4 in sera of 121 patients with allergic rhinitis and 36 non-allergic individuals. All subjects were skin prick tested with Bt-total extract and inhibition tests were performed for IgE, IgG1, and IgG4 using both extracts (Bt-total and Bt-Con-A). Skin prick test showed that 58% of the patients were sensitized to Bt (Bt+), with 52% reactive to both mites (Bt and Dp) and 6% to Bt only. A broad spectrum of proteins (14-152 kDa) was visualized in Bt-total and components >27 kDa for the Bt-Con-A extract. ELISA showed a similar profile of IgE, IgG1 and IgG4 levels in response to Bt-total and Bt-Con-A extracts in different groups, although Bt+ patients showed a lower IgG4 reactivity to Bt-Con-A extract. Specific IgG1 levels were higher in Bt+ patients than in control subjects, and IgG4 levels showed no significant difference among groups. ELISA inhibition showed a partial IgE and total IgG1 and IgG4 cross-reactivity with Dp extract for Bt-total and Bt-Con-A extracts. We conclude that Con-A-binding components isolated from Bt constitute major allergens and are involved in both allergen sensitization (IgE response) and homeostasis maintenance (IgG1 and IgG4 responses).
Assuntos
Alérgenos/imunologia , Antígenos de Dermatophagoides/imunologia , Autoanticorpos/imunologia , Concanavalina A/farmacologia , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Rinite Alérgica Perene/imunologia , Animais , Especificidade de Anticorpos , Antígenos de Plantas , Estudos de Casos e Controles , Reações Cruzadas , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Humanos , Ácaros/imunologia , Índice de Gravidade de DoençaRESUMO
Blomia tropicalis (Bt) and Dermatophagoides pteronyssinus (Dp) are the prevalent house dust mites in tropical countries and are associated with allergic diseases. Glycosylated antigens are highly immunogenic and involved in different pathologies. We evaluated the presence of IgE, IgG1, and IgG4 to concanavalin A-binding antigens (Bt-Con-A) isolated from Bt-total extract in sera of allergic and non-allergic subjects. Bt-total and Bt-Con-A extracts were evaluated by SDS-PAGE and ELISA for reacting with IgE, IgG1, and IgG4 in sera of 121 patients with allergic rhinitis and 36 non-allergic individuals. All subjects were skin prick tested with Bt-total extract and inhibition tests were performed for IgE, IgG1, and IgG4 using both extracts (Bt-total and Bt-Con-A). Skin prick test showed that 58 percent of the patients were sensitized to Bt (Bt+), with 52 percent reactive to both mites (Bt and Dp) and 6 percent to Bt only. A broad spectrum of proteins (14-152 kDa) was visualized in Bt-total and components >27 kDa for the Bt-Con-A extract. ELISA showed a similar profile of IgE, IgG1 and IgG4 levels in response to Bt-total and Bt-Con-A extracts in different groups, although Bt+ patients showed a lower IgG4 reactivity to Bt-Con-A extract. Specific IgG1 levels were higher in Bt+ patients than in control subjects, and IgG4 levels showed no significant difference among groups. ELISA inhibition showed a partial IgE and total IgG1 and IgG4 cross-reactivity with Dp extract for Bt-total and Bt-Con-A extracts. We conclude that Con-A-binding components isolated from Bt constitute major allergens and are involved in both allergen sensitization (IgE response) and homeostasis maintenance (IgG1 and IgG4 responses).
Assuntos
Humanos , Animais , Alérgenos/imunologia , Antígenos de Dermatophagoides/imunologia , Autoanticorpos/imunologia , Concanavalina A/farmacologia , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Rinite Alérgica Perene/imunologia , Especificidade de Anticorpos , Estudos de Casos e Controles , Reações Cruzadas , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Índice de Gravidade de DoençaRESUMO
Several studies have shown that the presence of IgE antibodies to house dust mites (HDM), particularly Dermatophagoides pteronyssinus (Dpt), is an important risk factor for asthma. Allergen immunotherapy is indicated for patients with IgE antibodies to clinically relevant allergens. The aims of this study were to analyze the levels of specific serum IgE to Der p 1 and Der p 2 allergens in mite-sensitized atopic patients and to compare them with both in vivo (skin prick test) and in vitro (IgE-ELISA) sensitizations to Dpt crude extract. Forty-seven atopic patients with allergic rhinitis with or without intermittent or persistent mild asthma and positive skin prick test (SPT) to Dpt total extract were studied. Thirty age-matched healthy subjects with negative SPT to HDM were included as controls. Levels of total IgE and Dpt-, Der p 1- and Der p 2-specific IgE were measured by ELISAs in SPT-positive atopic patients and SPT-negative control subjects. Among 47 symptomatic atopic patients, 27 (57.4%) were double positive IgE to Der p 1 and Der p 2 allergens, 3 (6.4%) were single positive IgE to Der p 1, 4 (8.5%) were single positive IgE to Der p 2, and 13 (27.6%) were double negative IgE to both allergens. There was a significant correlation between Der p 1- and Der p 2-specific IgE levels, but not between Der p 1- or Der p 2-IgE levels and SPT results. The double negative IgE patients had the smallest skin test reactions although they showed high mean levels of total serum IgE. Therefore, the knowledge of specific IgE levels to Der p 1 and Der p 2 major allergens might support physicians for indication or follow-up in mite-sensitized patients under allergen-specific immunotherapy. These approaches might be important for obtaining improved safety and efficacy of the current clinical practice of allergen immunotherapy.
Assuntos
Antígenos de Dermatophagoides/imunologia , Imunoglobulina E/sangue , Adolescente , Adulto , Animais , Antígenos de Dermatophagoides/efeitos adversos , Proteínas de Artrópodes , Cisteína Endopeptidases , Dermatophagoides pteronyssinus/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina E/imunologia , Pessoa de Meia-Idade , Testes CutâneosRESUMO
Indoor allergens are major causative agents in allergic disease development. Besides homes, public transport vehicles have been considered important mite and pet allergen reservoirs. Our recent studies on allergen exposure in automobiles showed that different allergen levels are found in private cars versus taxis. We quantified group 1 Dermatophagoides spp. (Der 1), Felis domesticus (Fel d 1), and Canis familiaris (Can f 1) allergen levels by ELISA in dust samples from 60 taxi and 60 private car upholstered seats. Mean levels of Der 1 and Fel d 1 were significantly higher in taxis than private cars. A significantly higher percentage of taxis (42%) harboring sensitizing levels of Der 1 compared to private cars (5%) was also found. In spite of the low mean Fel d 1 levels, comparison of the percentage of vehicles with moderate Fel d 1 levels showed a significant difference between taxis and private cars (43% vs. 20%). On the other hand, mean Can f 1 levels were significantly higher in private cars compared to taxis concomitant with a significantly higher percentage of private cars containing moderate Can f 1 levels than taxis (53% vs. 28%). We conclude that upholstered seats from Brazilian taxis but not private cars constitute an important mite allergen reservoir. Thus, additional effective measures for the reduction of allergen exposure in vehicles within the global allergen avoidance strategy should also be routinely accomplished to minimize the induction of sensitization and symptoms in allergic patients.
Assuntos
Antígenos de Dermatophagoides/isolamento & purificação , Automóveis , Reservatórios de Doenças , Alérgenos/análise , Animais , Antígenos de Dermatophagoides/análise , Antígenos de Plantas , Proteínas de Artrópodes , Brasil , Distribuição de Qui-Quadrado , Cisteína Endopeptidases , Glicoproteínas/análise , Humanos , Estatísticas não ParamétricasRESUMO
BACKGROUND: Allergens from house dust mites (HDMs), Dermatophagoides pteronyssinus and Blomia tropicalis are clinically relevant in atopic respiratory diseases in tropical countries. AIMS OF THE STUDY: To evaluate immunoglobulin (Ig)E, IgG1, and IgG4 antibody responses to B. tropicalis in Brazilian atopic patients. METHODS: About 110 patients with allergic rhinitis with/without asthma and 33 control subjects underwent skin prick testing (SPT) with HDM extracts, and their sera were tested for IgE and IgG subclass antibodies to D. pteronyssinus and B. tropicalis by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. RESULTS: Most patients (56%) had positive SPT to B. tropicalis extract (B. tropicalis+ group), although 51% were reactive to both B. tropicalis and D. pteronyssinus and 6% were sensitized to B. tropicalis only. IgE-ELISA detected 43%B. tropicalis positivity with high-specific IgE levels in B. tropicalis+ patients. Specific IgG4 levels were higher in B. tropicalis+ than B. tropicalis- groups and correlated with specific IgE levels. The IgG1 levels to B. tropicalis were higher in patients than controls. The major allergenic B. tropicalis components recognized by B. tropicalis+ patient sera were the 54, 66, and 68 kDa proteins. The IgG4-binding protein profiles closely resembled that of IgE. The IgG1 antibodies recognizing multiple B. tropicalis protein species were detected in sera of all three patient groups. CONCLUSIONS: A large percentage of our allergic patients are B. tropicalis+. They are more frequently sensitized to high-molecular weight (HMW) B. tropicalis components than the major low-molecular weight (11-15 kDa) allergens detected in other studies. The results suggest that HMW B. tropicalis antigenic components are potential candidates for evaluating allergen exposure and sensitization, and for immunotherapy treatment.
Assuntos
Poeira , Hipersensibilidade/imunologia , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Ácaros/imunologia , Adulto , Animais , Antígenos/química , Antígenos/imunologia , Asma/etiologia , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Humanos , Immunoblotting , Peso Molecular , Rinite Alérgica Perene/complicações , Rinite Alérgica Perene/imunologiaRESUMO
Mite allergen exposure has been widely related to sensitization and development of allergic diseases. This study intended to evaluate the degree of allergen exposure in Uberaba, Brazil, through the measurements of Der f 1 and Der p 1 allergen levels associated with the acarologic analysis in house dust samples. A total of 240 dust samples were collected from 60 houses through vacuuming sofas and bedding, during the months of March and July 2000. Indoor temperature and relative humidity were also measured. Mites were counted and identified under light microscopy and allergen levels were measured by two-site monoclonal antibody ELISAs. The major mite family was Pyroglyphidae (39.4%), having D. pteronyssinus as the most frequent species (15.6%), followed by D. farinae (12.3%) and E. maynei (7.9%). The family Glycyphagidae was less commonly found (4.8%), with Blomia tropicalis (4.4%) as its majoritary member. The highest levels of Der f 1 and Der p 1 allergens were found in bedding samples in March (31.7 and 0.9 microg/g of dust, respectively), with Der f 1 levels significantly higher than Der p 1 (p < 0.0001). There was a significant positive correlation between the mite number and allergen levels. These results indicate that Dermatophagoides sp are the most frequent mites in our region followed by E. maynei. Therefore, the knowledge of the local mite fauna would improve the means of investigating the association between allergen exposure and sensitization, allowing the addition of new mite extracts in diagnostic tests.