RESUMO
Tectochrysin, a flavonoid compound, can be isolated from propolis, Alpinia oxyphylla Miq, and Lychnophora markgravii. This study evaluated the efficacy of tectochrysin in the treatment of shrimp tropomyosin (ST)-induced mouse asthma. Mice were sensitized with intraperitoneal (i.p.) injection of ST together with aluminum hydroxide as an adjuvant to establish a mouse model of asthma. Mice were i.p.-treated daily with tectochrysin. IgE levels in plasma, Th2 cytokines from both bronchoalveolar lavage (BAL) fluid and splenocytes, and CD200R on basophils in peripheral blood were measured. Histological analyses of lung tissues and accumulation of leukocytes in BAL fluid were performed. Lung eosinophil peroxidase, catalase and glutathione peroxidase activities were examined. ST was found to markedly increase eosinophilic inflammation and Th2 response in mice. Tectochrysin treatment reduced the level of IgE in plasma, the percentage of eosinophils in total white blood cells in peripheral blood, the total number of cells in BAL fluid, and eosinophil peroxidase activity in lung tissues. Tectochrysin attenuated ST-induced infiltration of eosinophils and epithelial mucus secretion in lung tissues and suppressed the overproduction of Th2 cytokines (IL-4 and IL-5) in BAL fluid. Tectochrysin also attenuated Th2 cytokine (IL-4 and IL-5) production from antigen-stimulated murine splenocytes in vitro, decreased the expression of CD200R on basophils in peripheral blood of asthmatic mice and inhibited IL-4 secretion from IgE-sensitized RBL-2H3 cells. In addition, tectochrysin enhanced catalase and glutathione peroxidase activities in lung tissues. Our findings demonstrate that TEC ameliorates allergic airway inflammation by suppressing Th2 response and oxidative stress.
Assuntos
Antiasmáticos/farmacologia , Anti-Inflamatórios não Esteroides/farmacologia , Asma/tratamento farmacológico , Flavonoides/farmacologia , Hipersensibilidade/tratamento farmacológico , Estresse Oxidativo/efeitos dos fármacos , Células Th2/imunologia , Alérgenos/imunologia , Animais , Antiasmáticos/administração & dosagem , Anti-Inflamatórios não Esteroides/administração & dosagem , Asma/induzido quimicamente , Asma/imunologia , Asma/patologia , Basófilos/metabolismo , Catalase/metabolismo , Modelos Animais de Doenças , Eosinófilos/metabolismo , Feminino , Flavonoides/administração & dosagem , Glutationa Peroxidase/metabolismo , Hipersensibilidade/imunologia , Imunoglobulina E/sangue , Injeções Intraperitoneais , Camundongos Endogâmicos C57BL , Muco/efeitos dos fármacos , Hipersensibilidade a Frutos do Mar/tratamento farmacológico , Hipersensibilidade a Frutos do Mar/imunologia , Tropomiosina/imunologiaRESUMO
Colorectal cancer (CRC) is one of the most common malignancy and cause of cancer death worldwide, and it still remains a therapeutic challenge for western medicine. There is strong evidence that, in addition to genetic predispositions, environmental factors have also a substantial impact in CRC development. The risk of CRC is attributed, among others to dietary habits, alcohol consumption, whereas physical activity, food containing dietary fiber, dairy products, and calcium supplements have a protective effect. Despite progress in the available therapies, surgery remains a basic treatment option for CRC. Implementation of additional methods of treatment such as chemo- and/or targeted immunotherapy, improved survival rates, however, the results are still far from satisfactory. One of the reasons may be the lack of deeper understanding of the interactions between the tumor and different types of cells, including tumor infiltrating granulocytes. While the role of neutrophils is quite well explored in many cancers, role of eosinophils and basophils is often underestimated. As part of this review, we focused on the function of different granulocyte subsets in CRC, emphasizing the beneficial role of eosinophils and basophils, as well as dichotomic mode of neutrophils action. In addition, we addressed the current knowledge on cells of granulocyte origin, specifically granulocytic myeloid derived suppressor cells (Gr-MDSCs) and their role in development and progression of CRC.
Assuntos
Basófilos/metabolismo , Neoplasias Colorretais/metabolismo , Eosinófilos/metabolismo , Células Supressoras Mieloides/metabolismo , Neutrófilos/metabolismo , Basófilos/patologia , Neoplasias Colorretais/patologia , Neoplasias Colorretais/terapia , Eosinófilos/patologia , Humanos , Células Supressoras Mieloides/patologia , Neutrófilos/patologiaRESUMO
It was reported that the expression of Toll-like receptor (TLR) 9 may be related to Th2-type allergic inflammation including allergic rhinitis (AR). However, little is known about the expression of TLR9 in the basophils in AR. In the present study, the expression of TLR9 was examined by flow cytometry analysis, and the expression of TLR9 mRNA in KU812 was determined by quantitative real-time PCR. The results showed that the percentage of TLR9+ CCR3+ cells in blood granulocytes increased by 46% in patients with AR, but not in peripheral blood mononuclear cells (PBMCs). Allergens namely Dermatophagoide allergen extract (DAE) and Platanus pollen allergen extract (PPAE) upregulated the expression of TLR9 in CCR3+ granulocytes by 76% and 84%, respectively. DAE and PPAE also enhanced the proportions of TLR9+ CD123+ HLA-DR- cells and TLR9+ CCR3+ CD123+ HLA-DR- cells in granulocytes and PBMCs of patients with AR. In order to investigate the actions of allergens on basophils, KU812 cells were used. It was observed that all KU812 cells expressed TLR9, and the expression intensity of TLR9 in a single KU812 cell was elevated by CpG. IL-37, IL-31, IL-33, Artemisia sieversiana wild allergen extract (ASWAE), DAE, OVA and Der p 1 induced an increase in the expression of TLR9 mRNA and IL-6 production in KU812 cells. It was shown that the percentage of TLR9-expressing basophils increased in the blood of ovalbumin (OVA)-sensitized mice. In conclusion, an increased expression of TLR9 and the production of IL-6 in basophils implicate that the contribution of basophils to AR is likely via TLR9.
Assuntos
Basófilos/metabolismo , Pólen/imunologia , Pyroglyphidae/imunologia , Rinite Alérgica/imunologia , Receptor Toll-Like 9/metabolismo , Adulto , Alérgenos/imunologia , Animais , Linhagem Celular , Modelos Animais de Doenças , Feminino , Citometria de Fluxo , Regulação da Expressão Gênica , Granulócitos/metabolismo , Antígenos HLA-DR/imunologia , Humanos , Imunoglobulina E/sangue , Interleucina-13/metabolismo , Interleucina-6/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pessoa de Meia-Idade , Ovalbumina/imunologia , RNA Mensageiro/biossíntese , Rinite Alérgica/patologia , Receptor Toll-Like 9/genética , Regulação para Cima/genética , Adulto JovemRESUMO
In this study, I investigated the allergy suppressive effect of tea made from fig (Ficus carica L.) leaves. In the rat basophil cell line RBL-2H3, degranulation was significantly suppressed by treatment with fig tea at the same time as addition of IgE antibodies (sensitization). IgE bound to the cell surface was liberated in the medium depending on the treatment time with fig tea. Therefore, it was suggested that the mechanism of action of fig tea is promotion of dissociation of IgE from FcεRI receptors. Such a mechanism is novel in food materials. On oral administration to mice, fig tea showed an inhibitory effect on allergic dermatitis. Furthermore, in tests using an atopic dermatitis model in NC/Nga mice, continued administration of fig tea suppressed symptom exacerbation after antigen administration.Abbreviations: AD: atopic dermatitis; ß-Hex: ß-hexosaminidase; FCM: flow cytometory; OA: oral administration; TA: transdermal administration.
Assuntos
Antialérgicos/uso terapêutico , Complexo Antígeno-Anticorpo/efeitos dos fármacos , Dermatite Atópica/tratamento farmacológico , Ficus/química , Imunoglobulina E/imunologia , Fitoterapia/métodos , Extratos Vegetais/uso terapêutico , Folhas de Planta/química , Receptores de IgE/imunologia , Chás de Ervas , Animais , Antialérgicos/farmacologia , Basófilos/metabolismo , Linhagem Celular , Modelos Animais de Doenças , Feminino , Imunoglobulina E/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Extratos Vegetais/farmacologia , Ratos , Receptores de IgE/metabolismo , Resultado do TratamentoRESUMO
BACKGROUND: Over 100 million people worldwide suffer from birch pollen allergy. Bet v 1 has been identified as the major birch pollen allergen. However, the molecular mechanisms of birch allergic sensitization, including the roles of Bet v 1 and other components of the birch pollen extract, remain incompletely understood. Here, we examined how known birch pollen-derived molecules influence the endolysosomal processing of Bet v 1, thereby shaping its allergenicity. METHODS: We analyzed the biochemical and immunological interaction of ligands with Bet v 1. We then investigated the proteolytic processing of Bet v 1 by endosomal extracts in the presence and absence of ligands, followed by a detailed kinetic analysis of Bet v 1 processing by individual endolysosomal proteases as well as the T-cell epitope presentation in BMDCs. RESULTS: We identified E1 phytoprostanes as novel Bet v 1 ligands. Pollen-derived ligands enhanced the proteolytic resistance of Bet v 1, affecting degradation kinetics and preferential cleavage sites of the endolysosomal proteases cathepsin S and legumain. E1 phytoprostanes exhibited a dual role by stabilizing Bet v 1 and inhibiting cathepsin protease activity. CONCLUSION: Bet v 1 can serve as a transporter of pollen-derived, bioactive compounds. When carried to the endolysosome, such compounds can modulate the proteolytic activity, including its processing by cysteine cathepsins. We unveil a paradigm shift from an allergen-centered view to a more systemic view that includes the host endolysosomal enzymes.
Assuntos
Alérgenos/imunologia , Antígenos de Plantas/imunologia , Endossomos/enzimologia , Peptídeo Hidrolases/metabolismo , Basófilos/imunologia , Basófilos/metabolismo , Betula/imunologia , Degranulação Celular/imunologia , Ativação Enzimática , Humanos , Imunoglobulina E/imunologia , Ligantes , Pólen/imunologia , Ligação Proteica , Proteínas RecombinantesRESUMO
In the present study, ethanol extracts of 90 wild mushroom samples from Nepal, and the pure compound hispidin, were screened for their ability to inhibit ß-hexosaminidase release (BHR) from rat basophilic leukemia-2H3 cells. Simultaneously, the toxicity of the extracts toward the cells was also determined, using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Samples belonging to the groups Hymenochaetales and Polyporales showed promising anti-allergic activity, with Phellinus adamantinus and Ganoderma lingzhi 3 allowing a mere 19.4% and 16.7% BHR, respectively, without any cell cytotoxicity. Moreover, the 50% inhibitory concentration (IC50) values for Inonotus clemensiae and P. adamantinus were determined to be 51.24 and 50.65 µg/mL, respectively; whereas hispidin, the major bioactive compound in I. clemensiae showed an IC50 value of 82.47 µg/mL. These findings are crucial in underscoring the medicinal value of the wild mushrooms of Nepal, as a source of strong antiallergic agents.
Assuntos
Antialérgicos/farmacologia , Basidiomycota/química , Produtos Biológicos/farmacologia , Pironas/farmacologia , Agaricales/química , Animais , Basófilos/metabolismo , Produtos Biológicos/química , Linhagem Celular Tumoral , Ganoderma/química , Leucemia Basofílica Aguda , Nepal , Ratos , beta-N-Acetil-Hexosaminidases/antagonistas & inibidoresRESUMO
BACKGROUND: Peach is a common elicitor of food allergic reactions. Peach-induced immediate reactions may occur as benign pollen-food syndromes, usually due to birch pollen-related PR-10 cross-reactivity in temperate climates, and as potentially severe primary food allergies, predominantly related to nsLTP Pru p 3 in Mediterranean regions. The newly described peach allergen Pru p 7 has gained recent attention as a potential peach allergy severity marker. Sensitization to Pru p 7 and its allergenic homologues of the gibberellin-regulated protein family occurs in areas with high Cupressaceae tree pollen exposure. OBJECTIVE: We sought to investigate the distribution, clinical characteristics and molecular associations of Pru p 7 sensitization among subjects with suspected peach allergy in different regions of France. METHODS: Subjects with suspected peach allergy (n = 316) were included. Diagnostic work-up was performed according to current guidelines, including open food challenge when required. IgE antibody measurements and competition experiments were performed using the ImmunoCAP assay platform. RESULTS: Sensitization to Pru p 7 was present in 171 (54%) of all subjects in the study and in 123 of 198 (62%) diagnosed as peach allergic, more than half of whom were sensitized to no other peach allergen. Frequency and magnitude of Pru p 7 sensitization were associated with the presence of peach allergy, the clinical severity of peach-induced allergic reactions and the level of cypress pollen exposure. Cypress pollen extract completely outcompeted IgE binding to Pru p 7. Pru p 7 was extremely potent in basophil activation tests. CONCLUSION AND CLINICAL RELEVANCE: A subtype of Cupressaceae pollinosis, characterized by Pru p 7 sensitization, can be an underlying cause of severe peach allergy.
Assuntos
Antígenos de Plantas/imunologia , Reações Cruzadas/imunologia , Cupressus/imunologia , Hipersensibilidade Alimentar/diagnóstico , Hipersensibilidade Alimentar/imunologia , Pólen/imunologia , Prunus persica/efeitos adversos , Adolescente , Adulto , Idoso , Alérgenos/imunologia , Basófilos/imunologia , Basófilos/metabolismo , Criança , Pré-Escolar , Suscetibilidade a Doenças , Feminino , Hipersensibilidade Alimentar/epidemiologia , Humanos , Imunização , Imunoglobulina E/imunologia , Lactente , Masculino , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , Índice de Gravidade de Doença , Adulto JovemRESUMO
BACKGROUND: Peanuts are most responsible for food-induced anaphylaxis in adults in developed countries. An effective and safe immunotherapy is urgently needed. The aim of this study was to investigate the immunogenicity, allergenicity, and immunotherapeutic efficacy of a well-characterized chemically modified peanut extract (MPE) adsorbed to Al(OH)3 . METHODS: Peanut extract (PE) was modified by reduction and alkylation. Using sera of peanut-allergic patients, competitive IgE-binding assays and mediator release assays were performed. The immunogenicity of MPE was evaluated by measuring activation of human PE-specific T-cell lines and the induction of PE-specific IgG in mice. The safety and efficacy of MPE adsorbed to Al(OH)3 was tested in two mouse models by measuring allergic manifestations upon peanut challenge in peanut-allergic mice. RESULTS: Compared to PE, the IgE-binding and capacity to induce allergic symptoms of MPE were lower in all patients. PE and MPE displayed similar immunogenicity in vivo and in vitro. In mice sensitized to PE, the threshold for anaphylaxis (drop in BT) upon subcutaneous challenge with PE was 0.01 mg, while at 0.3 mg MPE no allergic reaction occurred. Anaphylaxis was not observed when PE and MPE were fully adsorbed to Al(OH)3 . Both PE and MPE + Al(OH)3 showed to be efficacious in a model for immunotherapy. CONCLUSION: In our studies, an Al(OH)3 adsorbed MPE showed reduced allergenicity compared to unmodified PE, while the efficacy of immunotherapy is maintained. The preclinical data presented in this study supports further development of modified peanut allergens for IT.
Assuntos
Antígenos de Plantas/química , Antígenos de Plantas/imunologia , Arachis/química , Arachis/imunologia , Extratos Vegetais/química , Extratos Vegetais/imunologia , Anafilaxia/imunologia , Animais , Basófilos/imunologia , Basófilos/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Mediadores da Inflamação/metabolismo , Camundongos , Hipersensibilidade a Amendoim/imunologia , Linfócitos T/imunologia , Linfócitos T/metabolismoRESUMO
Background: Dietary non-digestible oligosaccharides (NDOs) have a protective effect against allergic manifestations in children at risk. Dietary intervention with NDOs promotes the colonization of beneficial bacteria in the gut and enhances serum galectin-9 levels in mice and atopic children. Next to this, NDOs also directly affect immune cells and low amounts may reach the blood. We investigated whether pre-incubation of whole blood from peanut-allergic patients with NDOs or galectin-9 can affect basophil degranulation. Methods: Heparinized blood samples from 15 peanut-allergic adult patients were pre-incubated with a mixture of short-chain galacto-oligosaccharides and long-chain fructo-oligosaccharides (scGOS/lcFOS), scFOS/lcFOS, or galectin-9 (1 or 5 µg/mL) at 37°C in the presence of IL-3 (0.75 ng/mL). After 2, 6, or 24 h, a basophil activation test was performed. Expression of FcεRI on basophils, plasma cytokine, and chemokine concentrations before degranulation were determined after 24 h. Results: Pre-incubation with scGOS/lcFOS, scFOS/lcFOS, or galectin-9 reduced anti-IgE-mediated basophil degranulation. scFOS/lcFOS or 5 µg/mL galectin-9 also decreased peanut-specific basophil degranulation by approximately 20%, mainly in whole blood from female patients. Inhibitory effects were not related to diminished FcεRI expression on basophils. Galectin-9 was increased in plasma after pre-incubation with scGOS/lcFOS, and both NDOs and 5 µg/mL galectin-9 increased MCP-1 production. Conclusion and clinical relevance: The prebiotic mixture scFOS/lcFOS and galectin-9 can contribute to decreased degranulation of basophils in vitro in peanut-allergic patients. The exact mechanism needs to be elucidated, but these NDOs might be useful in reducing allergic symptoms.
Assuntos
Alérgenos/imunologia , Arachis/efeitos adversos , Basófilos/imunologia , Degranulação Celular/imunologia , Imunomodulação , Oligossacarídeos/imunologia , Hipersensibilidade a Amendoim/imunologia , Adolescente , Adulto , Basófilos/metabolismo , Biomarcadores , Citocinas/metabolismo , Suplementos Nutricionais , Feminino , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Masculino , Pessoa de Meia-Idade , Hipersensibilidade a Amendoim/diagnóstico , Receptores de IgE/metabolismo , Adulto JovemRESUMO
BACKGROUND: Basophilic granulocytes (BGs) not only initiate the induction of Th2 cell differentiation, but also amplify the ongoing Th2 response. Shuang-Huang-Lian (SHL) is clinically used for relieving type I hypersensitivity by continuous treatment for several weeks. METHODS: ELISA, flow cytometry, magnetic activated cell sorting, isoelectric precipitation, hybridoma technique, transfection and luciferase reporter assay were used in this study. The statistical analysis was performed using a one-way ANOVA. RESULTS: Our recently published study demonstrated that SHL exerted a remarkable effect on mast cell stabilization. Herein, we sought to elucidate the effect of SHL on shrimp tropomyosin (ST)-induced Th2 immunity and its underlying mechanisms. The obtained data showed that continuous treatment with SHL significantly suppressed ST-stimulated Th2-cytokines release and IgE synthesis. A mechanistic study indicated that SHL not only reduced BG early IL-4 release before ST-specific IgE (sIgE) production, but also inhibited BG activation in the presence of sIgE, including suppressing CD200R surface expression and decreasing IL-4 production. Moreover, SHL markedly decreased the cytosolic Ca2+ (Ca2+[c]) level and inhibited the nuclear factor of activated T cells (NFAT) activation in RBL-2H3 cells. CONCLUSIONS: Collectively, SHL potently reduces ST-induced Th2 immunity by inhibiting the BG Ca2+-NFAT pathway and, thus, suppressing the early IL-4 release before sIgE synthesis and inhibiting BG activation in the presence of sIgE. This study provides the pharmacological basis for the clinical use of SHL to relieve type I hypersensitivity by a successive dose regimen.
Assuntos
Basófilos/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Células Th2/efeitos dos fármacos , Animais , Basófilos/metabolismo , Linhagem Celular Tumoral , Citocinas/análise , Citocinas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ratos , Células Th2/metabolismoRESUMO
BACKGROUND: Overlapping seasons and cross-reactivity, especially to grass pollen profilin, can hamper the diagnosis of birch pollen allergy. To identify the primary sensitizing allergen and the clinical relevance of cross-sensitization, we correlated sensitization profiles with in vitro and in vivo tests, symptom scores, and pollen counts. METHODS: A total of 433 patients with positive skin prick test (SPT) to birch pollen were analyzed regarding IgE to major birch and grass pollen allergens Bet v 1 and Phl p 1/p 5 and the profilins Bet v 2 and Phl p 12. Subgroups were analyzed by basophil activation test (BAT) and CAP-FEIA-based cross- and self-inhibition tests. RESULTS: A total of 349 patients were sensitized to Bet v 1, 44 patients to both Bet v 1 and Bet v 2, and 15 patients to Bet v 2 only. From Bet v 2-sensitized patients, 40 were also sensitized to Phl p 12. Ex vivo, Bet v 2 and Phl p 12 induced dose-dependent activation in basophils of these patients. Cross- and self-inhibition tests with both allergens confirmed cross-reactivity. However, semiquantitative analysis of SPTs demonstrated markedly increased reactivity to grass compared to birch pollen extract in Bet v 2 only sensitized patients. Accordingly, in most of those patients, clinical symptoms precisely correlated with grass pollen counts. CONCLUSION: Identification of the clinically relevant and sensitizing allergen needs correlation of actual pollen counts with clinical symptoms and sensitization status to major allergens. Semiquantitative analysis of SPT or BAT and determining profilin-specific IgE can contribute to making the diagnosis.
Assuntos
Alérgenos/imunologia , Betula/imunologia , Reações Cruzadas/imunologia , Pólen/imunologia , Profilinas/imunologia , Rinite Alérgica Sazonal/diagnóstico , Rinite Alérgica Sazonal/imunologia , Adolescente , Adulto , Especificidade de Anticorpos/imunologia , Basófilos/imunologia , Basófilos/metabolismo , Feminino , Humanos , Imunização , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Contagem de Leucócitos , Masculino , Rinite Alérgica Sazonal/sangue , Testes Cutâneos , Adulto JovemRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Oak bark has been used since ancient times in Europaen ethnomedicine because of its adstringent, antimicrobial and hemostatic features, e.g. as a remedy for the treatment of wounds and skin diseases. PURPOSE: Oak bark tannins are considered as bioactive natural products, interacting with surface proteins of mucous membranes and might be beneficial for the treatment of allergic diseases. This study investigated the effect of an oak bark decoction (OBD) and isolated tannin fractions on the degranulation capacity and cytokine/chemokine release from rat basophilic cells and human mast cells in vitro, which are essential for the initiation of early- and late-phase allergic reactions. METHODS AND METHODS: By chromatographic separation on Sephadex® LH-20 high- and low-molecular weight tannins were separated from OBD and the tannin composition analyzed by HPLC(DAD)-MSn. Then, the OBD and its fractions were tested in degranulation (ß-hexosaminidase activity) of allergen-specific-activated basophilic cells in a photometric assay. RESULTS: The OBD and the high-molecular tannin fraction showed a dose-dependent inhibition of cell degranulation. Furthermore, the OBD and particularly its high molecular weight tannin fraction exhibited an inhibitory activity on the IL-8-, IL-6- and TNF-α-secretion from stimulated human mast cells, detected and quantified by ELISA. CONCLUSION: The OBD and its high-molecular weight tannins revealed an impact on allergic mediator release of basophilic cells and human mast cells and thereby provide a rationale for the topical treatment with OBD preparations.
Assuntos
Alérgenos/metabolismo , Basófilos/efeitos dos fármacos , Degranulação Celular/efeitos dos fármacos , Mastócitos/efeitos dos fármacos , Casca de Planta/química , Extratos Vegetais/farmacologia , Quercus/química , Animais , Basófilos/metabolismo , Cromatografia Líquida , Humanos , Ratos , Espectrometria de Massas em TandemRESUMO
INTRODUCTION Allergic rhinitis (AR) is the most common atopic disease. Specific immunotherapy (SIT) is the only effective treatment method for AR. In uncertain diagnostic cases, before establishing eligibility for SIT, nasal provocation tests (NPTs) should be performed. However, there are numerous contraindications to performing NPTs, and there is ongoing search for an alternative in vitro method. OBJECTIVES The aim of the study was to determine whether a specific in vitro provocation, that is, the basophil activation test (BAT), may replace a specific in vivo provocation, that is, the NPT, in establishing patient's eligibility for SIT. PATIENTS AND METHODS The study included 30 patients with AR caused by allergy to house dust mite or birch pollen, referred for SIT. The assessment of basophil activation by measuring CD63 antigen expression was performed using the Flow2 CAST test. Basophils were stimulated with allergen preparation (concentrations of 5000, 500, and 50 standardized biological units) used in NPTs. BAT results were expressed as stimulation index (SI) and basophil reactivity (BR). RESULTS Allergen concentrations of 500 and 50 SBU proved to be appropriate for basophil stimulation. Median SI and BR were higher for positive NPT results than for negative NPT results (P <0.001). Sensitivity for SI and BR was in the range from 83% to 100%; specificity, from 78% to 89%; positive predictive value, from 75% to 87%; and negative predictive value, from 89% to 100%. We observed a high correlation of the analyzed parameters for the allergen concentrations of 500 and 50 SBU (range, 0.58-0.74; P <0.05). CONCLUSIONS If there are contraindications to performing the NPT, BAT may be regarded as an alternative in establishing patients' eligibility for SIT. The optimal concentrations of allergen preparations are 500 and 50 SBU. Both SI and BR are good indicators of basophil activation.
Assuntos
Testes Imunológicos , Imunoterapia , Testes de Provocação Nasal , Rinite Alérgica/diagnóstico , Adulto , Alérgenos , Animais , Basófilos/metabolismo , Feminino , Humanos , Masculino , Pólen/imunologia , Pyroglyphidae/imunologia , Rinite Alérgica/terapia , Sensibilidade e Especificidade , Tetraspanina 30/genética , Regulação para CimaRESUMO
BACKGROUND: We have developed a recombinant B cell epitope-based vaccine (BM32) for allergen-specific immunotherapy (AIT) of grass pollen allergy. The vaccine contains recombinant fusion proteins consisting of allergen-derived peptides and the hepatitis B surface protein domain preS as immunological carrier. METHODS: We conducted a randomized, double-blind, placebo-controlled AIT study to determine safety, clinical efficacy and immunological mechanism of three subcutaneous injections of three BM32 doses adsorbed to aluminum hydroxide versus aluminum hydroxide (placebo) applied monthly to grass pollen allergic patients (n=70). Primary efficacy endpoint was the difference in total nasal symptom score (TNSS) through grass pollen chamber exposure before treatment and 4weeks after the last injection. Secondary clinical endpoints were total ocular symptom score (TOSS) and allergen-specific skin response evaluated by titrated skin prick testing (SPT) at the same time points. Treatment-related side effects were evaluated as safety endpoints. Changes in allergen-specific antibody, cellular and cytokine responses were measured in patients before and after treatment. RESULTS: Sixty-eight patients completed the trial. TNSS significantly decreased with mean changes of -1.41 (BM32/20µg) (P=0.03) and -1.34 (BM32/40µg) (P=0.003) whereas mean changes in the BM32/10µg and placebo group were not significant. TOSS and SPT reactions showed a dose-dependent decrease. No systemic immediate type side effects were observed. Only few grade 1 systemic late phase reactions occurred in BM32 treated patients. The number of local injection site reactions was similar in actively and placebo-treated patients. BM32 induced highly significant allergen-specific IgG responses (P<0.0001) but no allergen-specific IgE. Allergen-induced basophil activation was reduced in BM32 treated patients and addition of therapy-induced IgG significantly suppressed T cell activation (P=0.0063). CONCLUSION: The B cell epitope-based recombinant grass pollen allergy vaccine BM32 is well tolerated and few doses are sufficient to suppress immediate allergic reactions as well as allergen-specific T cell responses via a selective induction of allergen-specific IgG antibodies. (ClinicalTrials.gov number, NCT01445002.).
Assuntos
Alérgenos/imunologia , Dessensibilização Imunológica , Epitopos de Linfócito B/imunologia , Poaceae/efeitos adversos , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Rinite Alérgica Sazonal/terapia , Vacinas/imunologia , Adulto , Alérgenos/química , Sequência de Aminoácidos , Antígenos de Plantas/química , Antígenos de Plantas/imunologia , Basófilos/imunologia , Basófilos/metabolismo , Degranulação Celular/imunologia , Dessensibilização Imunológica/efeitos adversos , Dessensibilização Imunológica/métodos , Epitopos de Linfócito B/química , Feminino , Humanos , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Ativação Linfocitária/imunologia , Masculino , Pessoa de Meia-Idade , Modelos Moleculares , Conformação Proteica , Proteínas Recombinantes de Fusão/imunologia , Rinite Alérgica Sazonal/diagnóstico , Pele/imunologia , Linfócitos T/imunologia , Vacinas/administração & dosagem , Vacinas/efeitos adversos , Adulto JovemRESUMO
BACKGROUND: α-melanocyte-stimulating hormone (α-MSH) was shown to inhibit allergic airway inflammation and exert suppressive effects on human basophils. OBJECTIVE: This study aims to extend our current knowledge on the melanocortin 1 receptor (MC1R) expression in nasal tissue of patients with allergic rhinitis (AR) and functional effects of α-MSH in human basophils especially from patients with allergic rhinitis. METHODS: MC1R expression before and after nasal allergen provocation was studied in nasal mucosal tissue of AR patients and in a mouse model of allergic airway inflammation using immunofluorescence. In vitro regulation of the MC1R and CD203c surface expression on whole-blood basophils of patients with AR and controls was assessed with flow cytometry. Functional effects of α-MSH on isolated basophils were analysed regarding apoptosis with flow cytometry and chemotaxis using a Boyden chamber assay. RESULTS: We detected an accumulation of MC1R-positive basophils in nasal mucosa tissue of patients with AR 24 h after nasal allergen provocation. Such accumulation was not present in mucosa sections from healthy controls. In mice with allergic airway inflammation, we found a clear accumulation of MC1R-positive basophils in the nasal tissue compared to control mice. MC1R expression was inducible in AR patients and controls by stimulation with anti-IgE. α-MSH inhibited anti-IgE and grass pollen induced upregulation of CD203c, but had no effect on chemotaxis or apoptosis of basophils in vitro. CONCLUSIONS AND CLINICAL RELEVANCE: MC1R-positive basophils accumulate in the nasal mucosa of patients with AR after nasal allergen provocation. Since α-MSH suppresses proinflammatory effector functions in human basophils via the MC1R, it constitutes an interesting novel target for modulating the allergic inflammatory response.
Assuntos
Receptor Tipo 1 de Melanocortina/metabolismo , Rinite Alérgica/imunologia , Rinite Alérgica/metabolismo , Adulto , Alérgenos/imunologia , Animais , Basófilos/imunologia , Basófilos/metabolismo , Biópsia , Quimiotaxia/imunologia , Modelos Animais de Doenças , Feminino , Expressão Gênica , Humanos , Imunoglobulina E/imunologia , Masculino , Camundongos , Mucosa Nasal/imunologia , Mucosa Nasal/metabolismo , Mucosa Nasal/patologia , Testes de Provocação Nasal , Diester Fosfórico Hidrolases/metabolismo , Pólen/imunologia , Pirofosfatases/metabolismo , Receptor Tipo 1 de Melanocortina/genética , Testes de Função Respiratória , Rinite Alérgica/diagnóstico , Rinite Alérgica/genética , Testes Cutâneos , Adulto Jovem , alfa-MSH/metabolismoRESUMO
BACKGROUND: Besides allergens, pollen release bioactive, low molecular weight compounds that modulate and stimulate allergic reactions. Clinical relevance of these substances has not been investigated to date. OBJECTIVE: To elucidate the effect of a non-allergenic, low molecular weight factors from aqueous birch pollen extracts (Bet-APE < 3 kDa) on the human allergic immune response in vivo. METHODS: Birch and grass pollen allergic individuals underwent skin prick testing with allergen alone, allergen plus Bet-APE < 3 kDa, or allergen plus pre-identified candidate substances from low molecular pollen fraction. Nasal allergen challenges were performed in non-atopic and pollen allergic individuals using a 3 day repeated threshold challenge battery. Subjects were either exposed to allergen alone or to allergen plus Bet-APE< 3 kDa. Local cytokine levels, nasal secretion weights, nasal congestion and symptom scores were determined. RESULTS: Skin prick test reactions to pollen elicited larger weals when allergens were tested together with the low molecular weight compounds from pollen. Similar results were obtained with candidate pollen-associated lipid mediators. In nasal lining fluids of allergic patients challenged with allergen plus Bet-APE < 3 kDa, IL-8 and IgE was significantly increased as compared to allergen-only challenged patients. These patients also produced increased amounts of total nasal secretion and reported more severe rhinorrhea than the allergen-only challenged group. CONCLUSIONS: Low molecular compounds from pollen enhance the allergen specific immune response in the skin and nose. They are therefore of potential clinical relevance in allergic patients.
Assuntos
Alérgenos/imunologia , Imunidade , Imunomodulação , Extratos Vegetais/imunologia , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Basófilos/imunologia , Basófilos/metabolismo , Betula/imunologia , Degranulação Celular/imunologia , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Mediadores da Inflamação/metabolismo , Interleucina-8/metabolismo , Peso Molecular , Mucosa Nasal/imunologia , Mucosa Nasal/metabolismo , Testes de Provocação Nasal , Extratos Vegetais/química , Pólen/química , Rinite Alérgica Sazonal/diagnóstico , Rinite Alérgica Sazonal/metabolismo , Testes Cutâneos , Avaliação de Sintomas , Células Th2/imunologia , Células Th2/metabolismoRESUMO
BACKGROUND: Phosphatidylinositol 3-kinase p110δ isoform (PI3K p110δ) activity is essential for mast cell activation, suggesting that inhibition of PI3K p110δ might be useful in treating allergic diseases. OBJECTIVE: We sought to determine the effect of the PI3K p110δ-selective inhibitor idelalisib on allergic responses. METHODS: This phase 1 randomized, double-blind, placebo-controlled, 2-period crossover study was conducted with the Vienna Challenge Chamber. Grass pollen-induced allergic symptoms were documented during screening. Eligible subjects received idelalisib (100 mg twice daily) or placebo for 7 days, with allergen challenge on day 7. After a 2-week washout period, subjects received the alternate treatment and repeated allergen challenge. Study measures included safety, nasal and nonnasal symptoms, nasal airflow, nasal secretions, basophil activation, and plasma cytokine levels. RESULTS: Forty-one patients with allergic rhinitis received idelalisib/placebo (n = 21) or placebo/idelalisib (n = 20). Idelalisib treatment was well tolerated. Mean total nasal symptom scores were lower during the combined idelalisib treatment periods compared with placebo (treatment difference [idelalisib - placebo], -1.78; 95% CI, -2.53 to -1.03; P < .001). Statistically significant differences were also observed for the combined treatment periods for total symptom scores, nasal airflow, nasal secretion weight, and nasal congestion scores. The percentage of ex vivo-activated basophils (CD63(+)/CCR3(+) cells; after stimulation with grass pollen) was substantially lower for idelalisib-treated compared with placebo-treated subjects. Plasma CCL17 and CCL22 levels were reduced after idelalisib treatment. CONCLUSION: Idelalisib treatment was well tolerated in patients with allergic rhinitis and appears to reduce allergic responses clinically and immunologically after an environmental allergen challenge.
Assuntos
Inibidores Enzimáticos/uso terapêutico , Inibidores de Fosfoinositídeo-3 Quinase , Purinas/uso terapêutico , Quinazolinonas/uso terapêutico , Rinite Alérgica/tratamento farmacológico , Adulto , Alérgenos/imunologia , Basófilos/imunologia , Basófilos/metabolismo , Inibidores Enzimáticos/farmacologia , Feminino , Volume Expiratório Forçado , Humanos , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Pólen/imunologia , Purinas/farmacologia , Quinazolinonas/farmacologia , Rinite Alérgica/diagnóstico , Rinite Alérgica/metabolismo , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: Venom immunotherapy is efficient to desensitize people suffering from insect sting allergies. However, the numerous injections required over several years and important risks of severe side reactions complicate the widespread use of immunotherapy. In the search for novel approaches to blunt the overwhelming pro-allergic Th2 response, we evaluated the therapeutic efficacy of a treatment based on a denatured form of the major allergen, phospholipase A2, associated with microbubbles (PLA2denat -MB) in a mouse model of honeybee venom allergy. METHODS: Antibodies measured by ELISA, T-cell responses assessed by CFSE-based proliferation assays and ELISA, and basophil degranulation were examined after PLA2denat -MB-based therapeutic treatment of sensitized mice. Mice were challenged with a lethal dose of PLA2 to evaluate protection against anaphylaxis. RESULTS: Therapeutic subcutaneous administration of two different PLA2denat -MB formulations, in contrast to PLA2denat alone, reduced allergic symptoms and protected all mice from anaphylaxis-mediated death after allergen challenge. At the functional level, the use of PLA2denat decreased IgE-mediated basophil degranulation as compared to the native form of the allergen. In comparison with PLA2denat alone, both PLA2denat -MB formulations decreased allergen-specific Th2 CD4 T-cell reactivity. At the mechanistic level, PLA2denat -MB containing 20% palmitic acid and PEG induced PLA2-specific IgA and increased Foxp3(+) Treg frequencies and TGF-ß production, whereas the formulation bearing 80% palmitic acid triggered the production of IFN-γ, IgG2a, and IgG3. CONCLUSIONS: In contrast to conventional PLA2 subcutaneous immunotherapy, the therapeutic administration of PLA2-MB treatment to mice that already had established allergy to PLA2 protects all subsequently challenged animals.
Assuntos
Alérgenos/imunologia , Venenos de Abelha/imunologia , Abelhas/imunologia , Hipersensibilidade/imunologia , Hipersensibilidade/terapia , Microbolhas , Fosfolipases A2/administração & dosagem , Anafilaxia/imunologia , Anafilaxia/terapia , Animais , Especificidade de Anticorpos/imunologia , Basófilos/imunologia , Basófilos/metabolismo , Degranulação Celular/imunologia , Citocinas/metabolismo , Dessensibilização Imunológica , Modelos Animais de Doenças , Feminino , Hipersensibilidade/mortalidade , Tolerância Imunológica , Imunoglobulina E/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Mordeduras e Picadas de Insetos/imunologia , Camundongos , Ratos , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismoRESUMO
For centuries, itch was categorized as a submodality of pain. Recent research over the last decade has led to the realization that itch is in fact a separate and distinct, albeit closely related, sensation. Chronic itch is a common complaint and has numerous etiologies. Various receptors (TRPA1, TRPV1, PAR2, gastrin-releasing peptide receptor (GRPR), Mas-related G proteins), secreted molecules (histamine, nerve growth factor (NGF), substance P (SP), proteases), and cytokines/chemokines (thymic stromal lymphopoietin (TSLP), IL-2, IL-4, IL-13, and IL-31) are implicated as mediators of chronic pruritus. While much remains unknown regarding the mechanisms of chronic itch, this much is certain: there is no singular cause of itch. Rather, itch is caused by a complex interface between skin, keratinocytes, cutaneous nerve fibers, pruritogenic molecules, and the peripheral and central nervous systems. Atopic dermatitis is one of the most itchy skin dermatoses and affects millions worldwide. The sensation of atopic itch is mediated by the interplay between epidermal barrier dysfunction, upregulated immune cascades, and the activation of structures in the central nervous system. Clinicians are in possession of an arsenal of different treatment options ranging from moisturizers, topical immunomodulators, topical anesthetic ion channel inhibitors, systemic immunomodulators, as well as oral drugs capable of reducing neural hypersensitization. Emerging targeted therapies on the horizon, such as dupilumab, promise to usher in a new era of highly specific and efficacious treatments. Alternative medicine, stress reduction techniques, and patient education are also important treatment modalities. This review will focus on the mediators of chronic pruritus mainly associated with atopic dermatitis (atopic itch), as well as numerous different therapeutic options.
Assuntos
Dermatite Atópica/diagnóstico , Dermatite Atópica/metabolismo , Mediadores da Inflamação/metabolismo , Prurido/metabolismo , Basófilos/imunologia , Basófilos/metabolismo , Terapia Combinada/efeitos adversos , Terapia Combinada/métodos , Citocinas/metabolismo , Dermatite Atópica/fisiopatologia , Dermatite Atópica/terapia , Descoberta de Drogas , Epiderme/inervação , Epiderme/metabolismo , Epiderme/patologia , Histamina/metabolismo , Humanos , Neuropeptídeos/metabolismo , Percepção , Prurido/fisiopatologia , Células Receptoras Sensoriais/metabolismo , Transdução de Sinais , Transmissão Sináptica , Resultado do Tratamento , Linfopoietina do Estroma do TimoRESUMO
Th2 type immune responses are essential for protective immunity against parasites and play crucial roles in allergic disorders. Helminth parasites secrete a variety of proteases for their infectious cycles including for host entry, tissue migration, and suppression of host immune effector cell function. Furthermore, a number of pathogen-derived antigens, as well as allergens such as papain, belong to the family of cysteine proteases. Although the link between protease activity and Th2 type immunity is well documented, the mechanisms by which proteases regulate host immune responses are largely unknown. Here, we demonstrate that the cysteine proteases papain and bromelain selectively cleave the α subunit of the IL-3 receptor (IL-3Rα/CD123) on the surface of murine basophils. The decrease in CD123 expression on the cell surface, and the degradation of the extracellular domain of recombinant CD123 were dependent on the protease activity of papain and bromelain. Pre-treatment of murine basophils with papain resulted in inhibition of IL-3-IL-3R signaling and suppressed IL-3- but not thymic stromal lymphopoietin-induced expansion of basophils in vitro. Our unexpected findings illuminate a novel mechanism for the regulation of basophil functions by protease antigens. Because IL-3 plays pivotal roles in the activation and proliferation of basophils and in protective immunity against helminth parasites, pathogen-derived proteases might contribute to the pathogenesis of infections by regulating IL-3-mediated functions in basophils.