RESUMO
OBJECTIVE: We investigated the best strategy for adult asthmatics to avoid exposure to Dermatophagoides group (Der-1) allergens. METHODS: Adult atopic asthmatics (n = 111) followed a 32-item checklist for avoiding Der-1 allergen exposure. Twenty-five patients were excluded through incomplete sampling; 50 remaining patients encased their pillows/futons/mattresses in microfine-fiber covers, 13 used vacuum cleaners with dust-mite-collection nozzles, and 23 acted as non-intervention controls. During August-October 2010 and August-October 2011, dust samples were collected in Petri dishes placed in bedrooms for 2 weeks and from mattresses/futons by using adhesive tape on one morning. A Der-1 level decrease was defined as a mean 2011 Der-1 level of <1 as a ratio of the 2010 level on tape or Petri dish samples. We analyzed the associations between Der-1 level change (by ELISA) and % weekly variability in peak expiratory flow (PEF) or fraction of exhaled nitric oxide (FeNO) after intervention. RESULTS: Der-1 levels decreased significantly in the covers group but not the vacuuming group. FeNO levels and PEF variability were unchanged in both groups. In patients whose Petri dish or tape samples showed decreased Der-1 levels, the % PEF variability was lower in 2011 than in 2010, but FeNO levels were unchanged. Three interventions (vacuuming all family members' mattress/futon surfaces at least weekly or after exposure of the futons to sunlight, and floor wiping before vacuuming), plus using covers, were the most effective management strategy in reducing Der-1 levels. CONCLUSIONS: This environmental and bedding maintenance program may help manage adult atopic asthma.
Assuntos
Alérgenos/análise , Antígenos de Dermatophagoides/análise , Proteínas de Artrópodes/análise , Asma/prevenção & controle , Cisteína Endopeptidases/análise , Exposição Ambiental/análise , Hipersensibilidade Imediata/prevenção & controle , Adulto , Idoso , Poluição do Ar em Ambientes Fechados/análise , Poluição do Ar em Ambientes Fechados/prevenção & controle , Asma/metabolismo , Asma/fisiopatologia , Roupas de Cama, Mesa e Banho , Gerenciamento Clínico , Poeira/análise , Poeira/prevenção & controle , Exposição Ambiental/prevenção & controle , Feminino , Humanos , Hipersensibilidade Imediata/metabolismo , Hipersensibilidade Imediata/fisiopatologia , Imunoglobulina E/sangue , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/metabolismo , Pico do Fluxo ExpiratórioRESUMO
Prompt diagnosis of allergic bronchopulmonary mycosis (ABPM) is an important clinical issue in preventing irreversible lung damage. Therefore, a good serological marker for the diagnosis of ABPM is desired in clinical practice. The measurement of IgE antibody to crude Aspergillus fumigatus allergen is considered the first step in screening asthmatic patients for allergic bronchopulmonary aspergillosis (ABPA). However, presence of IgE to A. fumigatus does not always indicate genuine sensitization to A. fumigatus because of cross-reactivity between crude extracts from different fungal sources. The application of molecular-based allergy diagnosis can solve this problem. The specificity of testing can be greatly improved by measuring the IgE antibody to Asp f 1 and f 2, specific allergen components for genuine A. fumigatus allergy. The problem of cross-reactivity between crude fungal extracts is also true for the identification of genuine causal fungi in each ABPM patient. Some patients with ABPM induced by fungi other than Aspergillus may be consistent with ABPA diagnostic criteria because current criteria depend on IgE/IgG reactivity to crude extracts. Accurate identification of genuine causal fungi for ABPM is of clinical importance, considering that clinical presentation, anti-fungal treatment strategies and disease prognosis can be influenced by different causal fungi. The diagnosis of causal fungi can be robustly validated by the confirmation of genuine sensitization to fungi after measuring IgE to specific allergen components, as well as repeated microbiological isolation of the fungi from their airway.
Assuntos
Aspergilose Broncopulmonar Alérgica/diagnóstico , Aspergilose Broncopulmonar Alérgica/etiologia , Aspergillus fumigatus/imunologia , Testes Sorológicos , Alérgenos/imunologia , Anticorpos Antifúngicos/imunologia , Especificidade de Anticorpos/imunologia , Antígenos de Fungos/imunologia , Aspergillus fumigatus/genética , Reações Cruzadas/imunologia , Humanos , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Técnicas de Diagnóstico Molecular , Testes Sorológicos/métodosRESUMO
BACKGROUND: Cry j 2 and Cha o 2 are major allergens in Japanese cedar (Cryptomeria japonica; CJ) and Japanese cypress (Chamaecyparis obtusa; CO) pollen, respectively. Here, we assessed the epitopes related to the cross-reactivity between Cry j 2 and Cha o 2 using in vitro analyses. METHODS: Peptides were synthesized based on Cry j 2 sequential epitopes and relevant Cha o 2 amino acid sequences. Four representative monoclonal antibodies (mAbs) against Cry j 2 were used according to their epitope recognitions. Serum samples were collected from 31 patients with CJ pollinosis. To investigate cross-reactivity between Cry j 2 and Cha o 2, ELISA and inhibition ELISA were performed with mAbs and sera from patients with CJ pollinosis. RESULTS: Two of four mAbs had reactivity to both Cry j 2 and Cha o 2. Of these two mAbs, one mAb (T27) recognized the amino acid sequence (169)KVVNGRTV(176) on Cha o 2. This is related to the core epitope (169)KWVNGREI(176) on Cry j 2, which is an important IgE epitope. In addition, we found that these correlative sequences and purified allergens showed cross-reactivity between Cry j 2 and Cha o 2 in IgE of CJ patients. CONCLUSIONS: We demonstrated the importance of (169)KVVNGRTV(176) in Cha o 2 for cross-reactivity with the Cry j 2 epitope (169)KWVNGREI(176), which plays an important role in allergenicity in CJ pollinosis. Our results are useful for the development of safer and more efficient therapeutic strategies for the treatment of CJ and CO pollen allergies.
Assuntos
Antígenos de Plantas/imunologia , Reações Cruzadas/imunologia , Cryptomeria/imunologia , Cupressus/imunologia , Imunoglobulina E/imunologia , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Alérgenos/imunologia , Sequência de Aminoácidos , Anticorpos Monoclonais/imunologia , Ensaio de Imunoadsorção Enzimática , Epitopos/química , Epitopos/imunologia , Humanos , Peptídeos/química , Peptídeos/imunologia , Proteínas de Plantas/imunologia , Rinite Alérgica Sazonal/diagnóstico , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: In the 1990s, the Japanese Society of Allergology (JSA) standardized Japanese cedar pollen allergen vaccines. In the present study, the task force for house dust mite (HDM) allergen standardization of the Committee for Allergens and Immunotherapy of JSA reports the standardization of HDM allergen vaccines in Japan. METHODS: In vivo allergenic potency was determined by intradermal testing of 51 Japanese adults with positive serum specific IgE to HDM allergens. In vitro total IgE binding potency was analyzed by competition ELISA using a pooled serum, with sera obtained from 10 allergic patients. The amounts of HDM group 1 (Der 1) and group 2 major allergens in eight HDM allergen extracts were measured by sandwich ELISAs. Correlation between the in vitro total IgE binding potency and major allergen levels was analyzed. RESULTS: We selected a JSA reference HDM extract and determined its in vivo allergenic potency. The in vitro total IgE binding potency significantly correlated with Der 1 content, group 2 allergen content, and their combined amount, indicating that measurement of major allergen contents can be used as a surrogate in vitro assay. CONCLUSIONS: The task force determined the in vivo allergenic potency (100,000 JAU/ml) and Der 1 content (38.5 µg/ml) of the JSA reference HDM extract, selected the measurement of Der 1 content as the surrogate in vitro assay, and decided that manufacturers can label a HDM allergen extract as having a titer of 100,000 JAU/ml if it contains 22.2-66.7 µg/ml of Der 1.
Assuntos
Antígenos de Dermatophagoides/imunologia , Imunoterapia/normas , Vacinas/normas , Adulto , Alérgenos/imunologia , Feminino , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Japão , Masculino , Pessoa de Meia-Idade , Sociedades Médicas , Adulto JovemRESUMO
BACKGROUND: Limited information is available regarding the clinical usefulness of measuring the levels of IgE to allergen components from house dust mites (HDMs) in the diagnosis of genuine HDM allergy. METHODS: To evaluate the diagnostic usefulness of measuring levels of serum IgE antibodies (Abs) to allergen components from Dermatophagoides pteronyssinus (DP) as a predictor of immediate asthmatic response (IAR) to bronchoprovocation, we studied 55 DP-sensitized asthmatic patients who underwent a bronchoprovocation test using crude DP extract. The levels of IgE Abs to crude DP, nDer p 1, rDer p 2, and rDer p 10 in patients who showed IAR (n = 41) were compared with those in patients who showed no IAR (n = 14). RESULTS: While the frequencies of positivity for IgE Abs to nDer p 1 and rDer p 2 among the entire study population were 89 and 86%, respectively, all patients with IAR tested positive for both of them with high IgE concentrations. The areas under the receiver operating characteristic curves for IgE to nDer p 1 and rDer p 2 as predictors of IAR were 0.913 and 0.906, respectively. The specificity of IgE to nDer p 1 and rDer p 2 was higher than IgE to crude DP even at low cut-off points. CONCLUSIONS: IgE to nDer p 1 and/or rDer p 2 was highly predictive of allergen-induced IAR. These findings validate the clinical usefulness of measuring the levels of IgE to nDer p 1 and rDer p 2 as a diagnostic tool for genuine HDM allergy.
Assuntos
Alérgenos/imunologia , Antígenos de Dermatophagoides/imunologia , Proteínas de Artrópodes/imunologia , Cisteína Endopeptidases/imunologia , Dermatophagoides pteronyssinus/imunologia , Hipersensibilidade/diagnóstico , Hipersensibilidade/imunologia , Imunoglobulina E/imunologia , Adulto , Animais , Biomarcadores , Testes de Provocação Brônquica , Feminino , Volume Expiratório Forçado , Humanos , Imunoglobulina E/sangue , Masculino , Pessoa de Meia-Idade , Curva ROCRESUMO
BACKGROUND: In the 1990s, the Japanese Society of Allergology (JSA) standardized Japanese cedar pollen allergen vaccines. In the present study, the task force for house dust mite (HDM) allergen standardization of the Committee for Allergens and Immunotherapy of JSA reports the standardization of HDM allergen vaccines in Japan. METHODS: In vivo allergenic potency was determined by intradermal testing of 51 Japanese adults with positive serum specific IgE to HDM allergens. In vitro total IgE binding potency was analyzed by the competitive ELISA using a pooled serum, with sera obtained from 10 allergic patients. Concentrations of HDM group 1 (Der 1) and group 2 major allergens in eight HDM allergen extracts were measured by sandwich ELISAs. Correlation between the in vitro total IgE binding potency and major allergen levels was analyzed. RESULTS: We selected a JSA reference HDM extract and determined its in vivo allergenic potency. The in vitro total IgE binding potency significantly correlated with Der 1 content, group 2 allergen content, and their combined amount, indicating that measurement of major allergen contents can be used as a surrogate in vitro assay. CONCLUSIONS: The task force determined the in vivo allergenic potency (100000 JAU/ml) and Der 1 content (38.5 µg/ml) of the JSA reference HDM extract, selected the measurement of Der 1 content as the surrogate in vitro assay, and decided that manufacturers can label a HDM allergen extract as having a titer of 100000 JAU/ml if it contains 22.2-66.7 µg/ml of Der 1.
Assuntos
Alérgenos/imunologia , Hipersensibilidade/imunologia , Pyroglyphidae/imunologia , Vacinação/normas , Vacinas/imunologia , Adulto , Animais , Feminino , Humanos , Imunoglobulina E/imunologia , Técnicas In Vitro , Testes Intradérmicos , Masculino , Sociedades MédicasRESUMO
PURPOSE: Japanese cedar (Cryptomeria japonica; CJ) pollinosis is a type I allergy induced by CJ pollen, and Cry j 2 is one of the major allergens in this pollen. In a previous study, we analyzed IgE epitopes on Cry j 2 in humans by using synthetic peptides. The main purpose of this study was to identify B-cell epitopes on Cry j 2 in patients with CJ pollinosis by using monoclonal antibodies (mAbs) for Cry j 2. METHODS: We used ELISA with mAbs for the epitope analysis. Sera samples were collected from 80 patients with CJ pollinosis, and allergenic epitopes for mAbs and human IgE were identified using ELISA with synthetic peptides. The importance of the epitopes for human IgE was analyzed using an inhibition ELISA. RESULTS: Four independent epitopes (epitope #1, #2, #3, and #4) were identified on Cry j 2 with the use of mAbs. Epitope #3 and #4, corresponding to peptides No. 25 and No. 33, respectively, were newly determined as epitopes for mAbs and human IgE. Inhibition ELISA showed that not only epitope #2 (sequential) but epitope #1 (conformational) may play an important role in the CJ pollinosis. CONCLUSIONS: Our results revealed 4 epitopes, including two new ones, on Cry j 2. We also found that inhibition ELISA with appropriate mAbs could be a viable method of evaluating the importance of the conformational and sequential epitopes for human IgE. These results are beneficial for the development of safer and more efficient therapeutic strategies for treating CJ pollinosis.
Assuntos
Alérgenos/imunologia , Anticorpos Monoclonais/imunologia , Cryptomeria/imunologia , Imunoglobulina E/imunologia , Proteínas de Plantas/imunologia , Pólen/imunologia , Sequência de Aminoácidos , Antígenos de Plantas/imunologia , Epitopos de Linfócito B/imunologia , Humanos , Dados de Sequência Molecular , Peptídeos/imunologiaRESUMO
There are controversial reports on the relationship between helminthic infection and allergic diseases. Although IgE cross-reactivity between nematode Ascaris antigens and house dust-mite allergens in allergic patients have been reported, whether Ascaris or the mite is the primary sensitizer remains unknown. Here we found that immunization of naïve animals with Ascaris lumbricoides (Al) antigens induced production of antibodies cross-reactive to mite antigens from Dermatophagoides farinae (Df). Sera from Bangladeshi children showed IgE reactivity to Ascaris and mite extracts. IgG from rabbits immunized with Al extract exhibited reactivity to Df antigens. Treatment of the anti-Al antibody with Df antigen-coupled beads eliminated the reactivity to Df antigens. In immunoblot analysis, an approximately 100-kDa Df band was the most reactive to anti-Al IgG. The present study is the first step towards the establishment of animal models to study the relationship between Ascaris infection and mite-induced allergic diseases.
Assuntos
Alérgenos/imunologia , Anticorpos Anti-Helmínticos/imunologia , Antígenos de Dermatophagoides/imunologia , Antígenos de Helmintos/imunologia , Ascaris lumbricoides/imunologia , Dermatophagoides farinae/imunologia , Hipersensibilidade/prevenção & controle , Animais , Anticorpos Anti-Helmínticos/biossíntese , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/administração & dosagem , Ascaris lumbricoides/química , Reações Cruzadas , Dermatophagoides farinae/química , Hipersensibilidade/imunologia , Imunização , Imunoglobulina G/biossíntese , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , CoelhosRESUMO
BACKGROUND: Penicillium species are among the most common fungi present in the environment and are usually considered non-pathogenic to humans. However, in immunocompromised hosts they can be virulent pathogens and can cause death. Penicillium digitatum is a plant pathogen that commonly causes a postharvest fungal disease of citrus called green mould; it very rarely causes systemic mycosis in humans. Here, we report a case of fatal pneumonia due to P. digitatum infection, as confirmed by repeated examination of cultured sputum. CASE PRESENTATION: A cavity was found in the left upper lung on routine chest X-ray in a 78-year-old undernourished male who had been diagnosed at age 66 with bronchial asthma and pulmonary emphysema. No increased sputum production was present. The presence of antigen-specific precipitating antibodies to Aspergillus flavus and P. digitatum was confirmed in the patient's serum and also later pleural fluid by using Ouchterlony double immunodiffusion testing with A. flavus and P. digitatum antigens. The patient was treated over a period of months with itraconazole, micafungin, voriconazole, amphotericin B, and antibacterials. However, the cavity enlarged, the pleural effusion increased, and the patient began producing purulent sputum. He died from progressive renal failure. From sputum culture only one fungus was isolated repeatedly on potato-dextrose agar in large quantities. This fungus was confirmed to be P. digitatum by molecular identification. Partial sequences of the beta-tubulin gene were determined by using the primers Bt2a and Bt2b for PCR amplification and sequencing and underwent a BLAST search at the National Centre for Biotechnology Information, these results confirmed that the isolated fungus was P. digitatum. CONCLUSION: To our knowledge, this is the first report of pulmonary infection with P. digitatum. Our patient had pulmonary emphysema and was elderly, and undernourished. These factors might have facilitated the infection. In his case, antimycotics were ineffective in treating the lung involvement. Although human infection with P. digitatum is considered rare, it appears that this organism can be very virulent and resistant to antimycotics.
Assuntos
Micoses/microbiologia , Penicillium/isolamento & purificação , Pneumonia/microbiologia , Idoso , Evolução Fatal , Humanos , Masculino , Desnutrição/complicações , Micoses/complicações , Pneumonia/complicações , Enfisema Pulmonar/complicações , Escarro/microbiologiaRESUMO
BACKGROUND: Booklice (Liposcelis bostrichophila) are a common household insect pest distributed worldwide. Particularly in Japan, they infest 'tatami' mats and are the most frequently detected insect among all detectable insects, present at a frequency of about 90% in dust samples. Although it has been hypothesized that they are an important indoor allergen, studies on their allergenicity have been limited. METHODS: To clarify the allergenicity of booklice and the cross-reactivity of this insect allergen with allergens of other insects, patients sensitized to booklice were identified from 185 Japanese adults with allergic asthma using skin tests and IgE-ELISA. IgE-inhibition analysis, immunoblotting and immunoblotting-inhibition analysis were performed using sera from these patients. Allergenic proteins contributing to specific sensitization to booklice were identified by two-dimensional electrophoresis and two-dimensional immunoblotting. RESULTS: The booklouse-specific IgE antibody was detected in sera from 41 patients (22% of studied patients). IgE inhibition analysis revealed that IgE reactivity to the booklouse allergen in the sera from one third of booklouse-sensitized patients was not inhibited by preincubation with extracts from any other environmental insects in this study. Immunoblotting identified a 26-kD protein from booklouse extract as the allergenic protein contributing to specific sensitization to booklice. The amino acid sequence of peptide fragments of this protein showed no homology to those of previously described allergenic proteins, indicating that this protein is a new allergen. CONCLUSIONS: Sensitization to booklice was relatively common and specific sensitization to this insect not related to insect panallergy was indicated in this population.
Assuntos
Alérgenos/imunologia , Asma/epidemiologia , Asma/imunologia , Roupas de Cama, Mesa e Banho , Proteínas de Insetos/imunologia , Insetos/imunologia , Adulto , Alérgenos/isolamento & purificação , Animais , Asma/diagnóstico , Reações Cruzadas , Feminino , Humanos , Imunoglobulina E/sangue , Proteínas de Insetos/isolamento & purificação , Japão , Masculino , Prevalência , Fatores de Risco , Fatores Sexuais , Testes Cutâneos , FumarRESUMO
BACKGROUND: Reducing risk factors, such as exposure to allergens, and stepwise pharmacotherapy to achieve and maintain control of asthma are the mainstay of asthma care. The purpose of this study was to clarify the effect of exposure and sensitization to indoor allergens, including house dust mites, cats, and dogs, on the asthma control level. METHODS: Dust samples were collected from the mattresses of 101 adult asthma patient homes and the Dermatophagoides mite group 1 (Der 1), Fel d 1, and Can f 1 concentrations were measured using ELISA. Sensitization was determined by positive specific IgE antibodies. The Asthma Control Test (ACT), lowest peak expiratory flow (PEF) during 1 week expressed as a percentage of the highest PEF (Min%Max PEF), and spirometry were measured for the assessment of asthma control. Univariate and multivariate regression analyses were used to assess the relationships. RESULTS: Sixty-nine patients were exposed to high levels (>10µg/g dust for Der 1 and Can f 1 and >8µg/g dust for Fel d 1) of 1 or more allergens and 39 patients were sensitized to at least one allergen. Multivariate logistic regression analyses revealed that the FEV(1) (% of predicted value) was associated with low ACT scores (≤19) and that the number of highly exposed allergens and inhaled corticosteroid dose were associated with a low level of Min%Max PEF (<80%). CONCLUSIONS: The level of exposure to multiple indoor allergens, but not sensitization, is associated with the asthma control level determined by PEF variation.
Assuntos
Poluição do Ar em Ambientes Fechados/efeitos adversos , Alérgenos/imunologia , Asma/imunologia , Asma/prevenção & controle , Corticosteroides/administração & dosagem , Adulto , Idoso , Animais , Gatos , Cães , Feminino , Glicoproteínas/imunologia , Humanos , Imunoglobulina E/imunologia , Masculino , Pessoa de Meia-Idade , Pyroglyphidae/imunologia , Testes de Função Respiratória , Adulto JovemRESUMO
BACKGROUND: Several allergen sampling methods are available for the assessment of personal or indirect exposure to indoor allergens. As an index of exposure to inhalant allergens, assays of the amount of airborne allergens directly reflect personal exposure. OBJECTIVE: We evaluated the Petri dish sampling method of assessing the level of airborne Dermatophagoides dust mite group 1 (Der 1) allergens. METHODS: We collected settling dust samples from one person's bedroom over a period of 2 years by using a Petri dish, adhesive tape, and a vacuumed reservoir. We also collected settling dust samples from the bedrooms of 42 asthma patients by using a Petri dish and adhesive tape. The amounts of Der 1 collected on the Petri dishes and adhesive tapes were measured by sensitive fluorometric ELISA. RESULTS: Der 1 was detected in all samples by using a Petri dish. The mean coefficient of variation was approximately 15%. We found that Petri dishes set at lower sampling heights contained more Der 1 than those higher up. There were also seasonal changes in the amounts of Der 1 collected, with the highest amounts collected from summer to autumn, and the lowest amounts collected in winter. CONCLUSION: The Petri dish sampling method for collecting settling Der 1 is very simple and can be used as an alternative to personal air sampling, especially in large-scale studies.
Assuntos
Poluição do Ar em Ambientes Fechados/análise , Alérgenos/análise , Antígenos de Dermatophagoides/análise , Pyroglyphidae/imunologia , Animais , Monitoramento Ambiental/métodos , JapãoRESUMO
The aims of this study were to examine the therapeutic effects of sublingual immunotherapy (SLIT) and to identify potential biomarkers that would predict the therapeutic response in a randomized, double-blind, placebo-controlled clinical trial. The trial was carried out over two pollinosis seasons in 2007 and 2008. Carry-over therapeutic effects were analyzed in 2009. SLIT significantly ameliorated the symptoms of pollinosis during the 2008 and 2009 pollen seasons. Cry j 1-specific cytokine production in a subgroup of patients with mild disease in the SLIT group was significantly attenuated. The ratio of specific IgE to total IgE before treatment correlated with the symptom-medication score in the SLIT group in 2008. Patients with increased Cry j 1-iTreg in the SLIT group had significantly improved QOL and QOL-symptom scores. In summary, the specific IgE to total IgE ratio and upregulation of Cry j 1-iTreg are candidates for biomarker of the clinical response to SLIT.
Assuntos
Cryptomeria/fisiologia , Imunoglobulina E/sangue , Extratos Vegetais/imunologia , Pólen/imunologia , Rinite Alérgica Sazonal/terapia , Linfócitos T Reguladores/metabolismo , Administração Sublingual , Adolescente , Adulto , Idoso , Especificidade de Anticorpos , Biomarcadores , Citocinas/genética , Citocinas/metabolismo , Método Duplo-Cego , Feminino , Regulação da Expressão Gênica/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Extratos Vegetais/administração & dosagem , Adulto JovemRESUMO
BACKGROUND: Japanese cedar (Cryptomeria japonica) pollinosis is one of the most prevalent allergies in Japan. Recently, two reports described the positive effects of sublingual immunotherapy (SLIT) against Japanese cedar pollinosis. However, the therapeutic biomarkers for SLIT are still unclear. We performed this unblinded, nonrandomized, open-label study to identify therapeutic biomarkers for SLIT against Japanese cedar pollinosis. METHODS: We performed an open-label study during one pollinosis season in 2007, enrolling 19 patients from in-house volunteers suffering from Japanese cedar pollinosis. Peripheral blood was obtained from all participants before SLIT treatment as well as before and after the pollen season. The plasma levels of an immunoglobulin specific to a major allergen (Cry j 1) were determined. We analyzed the induction of regulatory T cells (iTregs), namely IL-10(+)Foxp3(+) cells in CD25(+)CD4(+) leukocytes, by flow cytometry. The Th2-type responses were analyzed by cytokine production from peripheral blood mononuclear cells after stimulation with Cry j 1. Clinical symptoms were estimated using a quality of life questionnaire in the middle of the pollen season. RESULTS: The difference in numbers of iTregs between the medium-only control cell culture and cells stimulat- ed with Cry j 1 was significantly decreased in the non-SLIT group but was unchanged in the SLIT group after the pollen season. The subgroup of the SLIT group with increased iTregs showed more attenuated Th2-type cytokine profiles, and symptom scores in the subgroup with increased iTregs were significantly lower than those in the subgroup with decreased iTregs. CONCLUSION: The antigen-specific iTreg level is a potential therapeutic biomarker that correlates with clinical pollinosis symptoms and may be involved in the therapeutic mechanisms of SLIT.
Assuntos
Dessensibilização Imunológica , Fatores de Transcrição Forkhead/biossíntese , Rinite Alérgica Sazonal/diagnóstico , Rinite Alérgica Sazonal/tratamento farmacológico , Linfócitos T Reguladores/metabolismo , Administração Sublingual , Adulto , Antígenos de Plantas/imunologia , Biomarcadores Farmacológicos/metabolismo , Antígenos CD4/biossíntese , Contagem de Células , Células Cultivadas , Cryptomeria , Progressão da Doença , Feminino , Humanos , Imunoglobulina E/sangue , Interleucina-10/biossíntese , Subunidade alfa de Receptor de Interleucina-2/biossíntese , Masculino , Pessoa de Meia-Idade , Proteínas de Plantas/imunologia , Rinite Alérgica Sazonal/imunologia , Rinite Alérgica Sazonal/patologia , Rinite Alérgica Sazonal/fisiopatologia , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/patologiaRESUMO
AIM: In order to specifically quantify the two major Dermatophagoides spp. allergens, Der p 1 and Der f 1, separately, we tried to establish a highly sensitive enzyme-linked immunosorbent assay (ELISA). METHODS: Ninety-six-well ELISA plates were coated with mouse monoclonal antibodies specific against Der p 1 or Der f 1. Allergen samples were incubated in the wells for 2 hours at 37 degrees C. After washing with PBS-T, biotinylated rabbit anti-Der 1 polyclonal antibody was added to the wells. The allergens were detected using horse radish peroxidase-conjugated streptavidin, an enzyme substrate (TMB/H2O2) and a microplate reader. RESULTS: The working range of both ELISA systems for Der p 1 and Der f 1 was 40-2500 pg/ml. The intra- and inter-assay coefficients of variation for reproducibility were 0.99-4.38% and 0.68-3.02%, respectively, in Der p 1 ELISA and 1.54-3.65% and 0.39-4.77%, respectively, in Der f 1 ELISA. Moreover, these ELISA systems showed that there was no cross-reactivity between Der p 1 and Der f 1 allergens. CONCLUSION: These ELISA systems may be useful for measuring less than 1 ng/ml of major mite allergens in house dust samples, various pharmaceutical studies such as evaluation of an allergen-inactivating agent, and standardizing recombinant/natural Dermatophagoides spp. allergens.
Assuntos
Alérgenos/análise , Antígenos de Dermatophagoides/análise , Ensaio de Imunoadsorção Enzimática/métodos , Proteínas de Artrópodes , Cisteína EndopeptidasesRESUMO
There has been much interest in the potential of using probiotic bacteria for treating allergic diseases. A double-blind, placebo-controlled study was conducted to examine the effectiveness of Lactobacillus GG (LGG) and L. gasseri TMC0356 (TMC0356) in alleviating Japanese cedar pollinosis (JCP), a seasonal allergic rhinitis caused by Japanese cedar pollen. Fermented milk prepared with the tested bacteria or placebo yoghurt was administered to 40 subjects with a clinical history of JCP for 10 weeks. Subjective symptoms, self-care measures and blood samples were compared between the two groups. Peripheral blood mononuclear cells (PBMCs) were collected from seven patients with JCP and in vitro cytokine production by the isolated PBMCs was analysed in the presence of heat-killed lactic acid bacteria. Consumption of the fermented milk significantly decreased the mean symptom score for nasal blockage after 9 weeks (P<0.05) and mean symptom-medication scores after 9 and 10 weeks when compared with the placebo group (P<0.01 and P<0.05 respectively). The tested strains of lactic acid bacteria affected cytokine production by isolated PBMCs in vitro in a strain-dependent manner. LGG significantly inhibited IL-4 and IL-5 production by PBMCs stimulated by both Cry j 1 and PHA. TMC0356 only suppressed IL-5 production stimulated by PHA. The fermented milk prepared with LGG and TMC0356 might be beneficial in JCP because of its effect on nasal blockage. The effects of LGG and TMC0356 might arise at least partly from their specific down-regulation of the human Th2 immune response.
Assuntos
Cryptomeria/imunologia , Citocinas/biossíntese , Lactobacillus/fisiologia , Leucócitos Mononucleares/metabolismo , Probióticos/farmacologia , Rinite Alérgica Sazonal/tratamento farmacológico , Adulto , Produtos Fermentados do Leite , Método Duplo-Cego , Feminino , Humanos , Imunoterapia , Interleucina-4/biossíntese , Interleucina-5/biossíntese , Masculino , Pólen/imunologia , Probióticos/uso terapêutico , Rinite Alérgica Sazonal/sangue , Rinite Alérgica Sazonal/imunologia , Especificidade da Espécie , Resultado do Tratamento , Iogurte/microbiologiaRESUMO
The prevalence of Japanese cedar pollinosis is high to 26.5%, and it is properly remarked as "national disease" in Japan. General therapy strategies are composed of self-care by using mask or glasses to avoid antigen, and medical-care by using second-generation antihistamines or nasal corticosteroid spray. Meanwhile, there are other so-called self-medication patients who utilize health foods but do not consult to any physician. Papilla, commercial name of one kind of those health foods, is a capsular packed with cedar pollen. In February 2007, we experienced a 49-year-old female patient who presented anaphylaxis shock after intake of Papilla capsular. In this case, her special-IgE antibody and histamine release test were both positive to cedar pollen. Although Cry j 1, the major allergen in the Papilla capsular is a small amount, the cause of anaphylaxis can not be reasoned as anything except the pollen contents in this product. In conclusion, it is appropriate to consider it exercise induce anaphylaxis shock due to playing tennis after intake of the capsular.
Assuntos
Anafilaxia/etiologia , Alimentos Orgânicos/efeitos adversos , Automedicação/efeitos adversos , Cedrus , Feminino , Humanos , Pessoa de Meia-Idade , PólenRESUMO
BACKGROUND: Japanese cedar pollen represents an important and unique allergen. Sublingual immunotherapy (SLIT) has been suggested to be a highly effective route of desensitization against a variety of allergens. However, little information is available about its use in cedar pollen allergy. METHODS: A blinded randomized, placebo-controlled trial employing SLIT for cedar pollinosis was conducted over a period of 6 months. Sixty-seven subjects were enrolled and the symptom scores during the pollen season were evaluated by a symptom diary, measurement of cedar-specific IgE and IgG4, and determination of Cry j-specific Th2 clones before SLIT and before and after the pollen season. RESULTS: No major adverse effects were observed in either group. The serum-specific IgG4 activity increased significantly after SLIT in the active group. The active group also exhibited significantly lower symptom scores compared to the placebo. The specific Th2 clone sizes were not significantly different between the groups before the pollen season. However, an increase in the clone size was observed after the pollen season in the placebo group, but not in the active group. CONCLUSION: Use of SLIT for Japanese cedar pollinosis was found to be safe and associated with an increase in cedar-specific IgG4 levels. Such therapy inhibited the increase in Cry j-specific Th2 clone size induced by pollen exposure. Finally, use of SLIT resulted in significant improvement of the clinical symptoms of cedar pollinosis in this patient population. These observations suggest that SLIT may offer another safe approach to the management of cedar pollinosis.
Assuntos
Alérgenos/administração & dosagem , Imunoterapia/métodos , Proteínas de Plantas/administração & dosagem , Rinite Alérgica Sazonal/terapia , Administração Sublingual , Adulto , Alérgenos/efeitos adversos , Alérgenos/imunologia , Antígenos de Plantas , Células Clonais/imunologia , Feminino , Citometria de Fluxo , Humanos , Imunoglobulina G/sangue , Interferon gama/sangue , Interleucina-4/sangue , Interleucina-4/imunologia , Interleucina-5/sangue , Interleucina-5/imunologia , Masculino , Proteínas de Plantas/efeitos adversos , Proteínas de Plantas/imunologia , Pólen/efeitos adversos , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Método Simples-Cego , Estatísticas não Paramétricas , Células Th2/imunologiaAssuntos
Antígenos de Plantas/imunologia , Hipersensibilidade Alimentar/diagnóstico , Glycine max/imunologia , Proteínas Recombinantes/imunologia , Rinite Alérgica Sazonal/diagnóstico , Adulto , Betula/imunologia , Reações Cruzadas , Feminino , Hipersensibilidade Alimentar/imunologia , Humanos , Imunização , Imunoglobulina E/sangue , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Rinite Alérgica Sazonal/imunologia , Testes CutâneosRESUMO
BACKGROUND: Airborne Cladosporium spp (Clad) antigens were measured by aeroallergen immnoblotting, and analyzed the seasonal fluctuation of the antigens. METHODS: Airborne Clad antigens (Clad) were transferred onto nitrocellulose membrane and treated with anti-Clad rabbit antiserum, then alkaline phosphatase-conjugated rabbit IgG. Finally, the spots from Clad antigens were visualized with BCIP/NBT substrate. RESULTS: Clear purple spots from Clad antigens were seen on the airborne samples. Two peaks of spring and autumn were observed on the study of daily examination of the spots. Many spots from Clad antigens were mainly observed from afternoon to midnight at daily basis. No correlation between the number of the spots and the four weather elements was seen. CONCLUSION: Spots from airborne Clad antigens can be obtained easily by use of an immunochemical technique. The great numbers of spores were seen in spring and autumn out of one year and from afternoon to midnight out of one day.