RESUMO
BACKGROUND: Common ragweed has been spreading as a neophyte in Europe. Elevated CO2 levels, a hallmark of global climate change, have been shown to increase ragweed pollen production, but their effects on pollen allergenicity remain to be elucidated. METHODS: Ragweed was grown in climate-controlled chambers under normal (380 ppm, control) or elevated (700 ppm, based on RCP4.5 scenario) CO2 levels. Aqueous pollen extracts (RWE) from control- or CO2 -pollen were administered in vivo in a mouse model for allergic disease (daily for 3-11 days, n = 5) and employed in human in vitro systems of nasal epithelial cells (HNECs), monocyte-derived dendritic cells (DCs), and HNEC-DC co-cultures. Additionally, adjuvant factors and metabolites in control- and CO2 -RWE were investigated using ELISA and untargeted metabolomics. RESULTS: In vivo, CO2 -RWE induced stronger allergic lung inflammation compared to control-RWE, as indicated by lung inflammatory cell infiltrate and mediators, mucus hypersecretion, and serum total IgE. In vitro, HNECs stimulated with RWE increased indistinctively the production of pro-inflammatory cytokines (IL-8, IL-1ß, and IL-6). In contrast, supernatants from CO2 -RWE-stimulated HNECs, compared to control-RWE-stimulated HNECS, significantly increased TNF and decreased IL-10 production in DCs. Comparable results were obtained by stimulating DCs directly with RWEs. The metabolome analysis revealed differential expression of secondary plant metabolites in control- vs CO2 -RWE. Mixes of these metabolites elicited similar responses in DCs as compared to respective RWEs. CONCLUSION: Our results indicate that elevated ambient CO2 levels elicit a stronger RWE-induced allergic response in vivo and in vitro and that RWE increased allergenicity depends on the interplay of multiple metabolites.
Assuntos
Ambrosia , Dióxido de Carbono , Alérgenos , Europa (Continente) , PólenRESUMO
BACKGROUND: Endotoxin (LPS) released from gram-negative bacteria causes strong immunologic and inflammatory effects and, when airborne, can contribute to respiratory conditions, such as allergic asthma. OBJECTIVES: We sought to identify the source of airborne endotoxin and the effect of this endotoxin on allergic sensitization. METHODS: We determined LPS levels in outdoor air on a daily basis for 4 consecutive years in Munich (Germany) and Davos (Switzerland). Air was sampled as particulate matter (PM) greater than 10 µm (PM > 10) and PM between 2.5 and 10 µm. LPS levels were determined by using the recombinant Factor C assay. RESULTS: More than 60% of the annual endotoxin exposure was detected in the PM > 10 fraction, showing that bacteria do not aerosolize as independent units or aggregates but adhered to large particles. In Munich 70% of annual exposure was detected between June 12th and August 28th. Multivariate modeling showed that endotoxin levels could be explained by phenological parameters (ie, plant growth). Indeed, days with high airborne endotoxin levels correlated well with the amount of Artemisia pollen in the air. Pollen collected from plants across Europe (100 locations) showed that the highest levels of endotoxin were detected on Artemisia vulgaris (mugwort) pollen, with little on other pollen. Microbiome analysis showed that LPS concentrations on mugwort pollen were related to the presence of Pseudomonas species and Pantoea species communities. In a mouse model of allergic disease, the presence of LPS on mugwort pollen was needed for allergic sensitization. CONCLUSIONS: The majority of airborne endotoxin stems from bacteria dispersed with pollen of only one plant: mugwort. This LPS was essential for inducing inflammation of the lung and allergic sensitization.
Assuntos
Poluição do Ar em Ambientes Fechados/análise , Lipopolissacarídeos , Material Particulado , Pólen , Animais , Artemisia/química , Artemisia/imunologia , Artemisia/microbiologia , Asma/imunologia , Asma/microbiologia , Alemanha , Humanos , Lipopolissacarídeos/química , Lipopolissacarídeos/imunologia , Camundongos Endogâmicos BALB C , Pantoea/imunologia , Material Particulado/química , Material Particulado/imunologia , Pólen/química , Pólen/imunologia , Pólen/microbiologia , Pseudomonas/imunologiaRESUMO
Ragweed pollen is the main cause of allergenic diseases in Northern America, and the weed has become a spreading neophyte in Europe. Climate change and air pollution are speculated to affect the allergenic potential of pollen. The objective of this study was to investigate the effects of NO2 , a major air pollutant, under controlled conditions, on the allergenicity of ragweed pollen. Ragweed was exposed to different levels of NO2 throughout the entire growing season, and its pollen further analysed. Spectroscopic analysis showed increased outer cell wall polymers and decreased amounts of pectin. Proteome studies using two-dimensional difference gel electrophoresis and liquid chromatography-tandem mass spectrometry indicated increased amounts of several Amb a 1 isoforms and of another allergen with great homology to enolase Hev b 9 from rubber tree. Analysis of protein S-nitrosylation identified nitrosylated proteins in pollen from both conditions, including Amb a 1 isoforms. However, elevated NO2 significantly enhanced the overall nitrosylation. Finally, we demonstrated increased overall pollen allergenicity by immunoblotting using ragweed antisera, showing a significantly higher allergenicity for Amb a 1. The data highlight a direct influence of elevated NO2 on the increased allergenicity of ragweed pollen and a direct correlation with an increased risk for human health.
Assuntos
Alérgenos/imunologia , Ambrosia/imunologia , Antígenos de Plantas/imunologia , Dióxido de Nitrogênio/farmacologia , Extratos Vegetais/imunologia , Poluição do Ar , Alérgenos/efeitos dos fármacos , Alérgenos/genética , Ambrosia/efeitos dos fármacos , Ambrosia/genética , Antígenos de Plantas/efeitos dos fármacos , Antígenos de Plantas/genética , Mudança Climática , Análise por Conglomerados , Eletroforese em Gel Bidimensional , Europa (Continente) , Humanos , Extratos Vegetais/genética , Proteínas de Plantas/efeitos dos fármacos , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , Estações do AnoRESUMO
BACKGROUND: Pollen released by allergenic members of the botanically unrelated families of Asteraceae and Cupressaceae represent potent elicitors of respiratory allergies in regions where these plants are present. As main allergen sources the Asteraceae species ragweed and mugwort, as well as the Cupressaceae species, cypress, mountain cedar, and Japanese cedar have been identified. The major allergens of all species belong to the pectate lyase enzyme family. Thus, we thought to investigate cross-reactivity pattern as well as sensitization capacities of pectate lyase pollen allergens in cohorts from distinct geographic regions. METHODS: The clinically relevant pectate lyase pollen allergens Amb a 1, Art v 6, Cup a 1, Jun a 1, and Cry j 1 were purified from aqueous pollen extracts, and patients' sensitization pattern of cohorts from Austria, Canada, Italy, and Japan were determined by IgE ELISA and cross-inhibition experiments. Moreover, we performed microarray experiments and established a mouse model of sensitization. RESULTS: In ELISA and ELISA inhibition experiments specific sensitization pattern were discovered for each geographic region, which reflected the natural allergen exposure of the patients. We found significant cross-reactivity within Asteraceae and Cupressaceae pectate lyase pollen allergens, which was however limited between the orders. Animal experiments showed that immunization with Asteraceae allergens mainly induced antibodies reactive within the order, the same was observed for the Cupressaceae allergens. Cross-reactivity between orders was minimal. Moreover, Amb a 1, Art v 6, and Cry j 1 showed in general higher immunogenicity. CONCLUSION: We could cluster pectate lyase allergens in four categories, Amb a 1, Art v 6, Cup a 1/Jun a 1, and Cry j 1, respectively, at which each category has the potential to sensitize predisposed individuals. The sensitization pattern of different cohorts correlated with pollen exposure, which should be considered for future allergy diagnosis and therapy.
Assuntos
Alérgenos/imunologia , Pólen/imunologia , Polissacarídeo-Liases/imunologia , Ambrosia/imunologia , Animais , Antígenos de Plantas/imunologia , Artemisia/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB CRESUMO
BACKGROUND: Allergies to grass pollen are the number one cause of outdoor hay fever. The human immune system reacts with symptoms to allergen from pollen. OBJECTIVE: We investigated the natural variability in release of the major group 5 allergen from grass pollen across Europe. METHODS: Airborne pollen and allergens were simultaneously collected daily with a volumetric spore trap and a high-volume cascade impactor at 10 sites across Europe for 3 consecutive years. Group 5 allergen levels were determined with a Phl p 5-specific ELISA in 2 fractions of ambient air: particulate matter of greater than 10 µm in diameter and particulate matter greater than 2.5 µm and less than 10 µm in diameter. Mediator release by ambient air was determined in FcεRI-humanized basophils. The origin of pollen was modeled and condensed to pollen potency maps. RESULTS: On average, grass pollen released 2.3 pg of Phl p 5 per pollen. Allergen release per pollen (potency) varied substantially, ranging from less than 1 to 9 pg of Phl p 5 per pollen (5% to 95% percentile). The main variation was locally day to day. Average potency maps across Europe varied between years. Mediator release from basophilic granulocytes correlated better with allergen levels per cubic meter (r(2) = 0.80, P < .001) than with pollen grains per cubic meter (r(2) = 0.61, P < .001). In addition, pollen released different amounts of allergen in the non-pollen-bearing fraction of ambient air, depending on humidity. CONCLUSION: Across Europe, the same amount of pollen released substantially different amounts of group 5 grass pollen allergen. This variation in allergen release is in addition to variations in pollen counts. Molecular aerobiology (ie, determining allergen in ambient air) might be a valuable addition to pollen counting.
Assuntos
Ar/análise , Basófilos/fisiologia , Material Particulado/análise , Proteínas de Plantas/análise , Pólen/química , Receptores de IgE/metabolismo , Rinite Alérgica Sazonal/imunologia , Degranulação Celular , Europa (Continente) , Humanos , Umidade , Poaceae/imunologia , Receptores de IgE/genética , Rinite Alérgica Sazonal/epidemiologia , Estações do AnoRESUMO
Pollen grains only represent a small fraction of the total amount of the viable biological particles present in the air, but pollen are the most important aeroallergens in the outdoor environment. The analysis of pollen has traditionally been carried out by microscopy, which can be traced back to the 17th century. Modern advances in molecular analysis could improve information for allergy sufferers and health care professionals. Pollen allergy (pollinosis) was first described in the 19th century. The prevalence of respiratory diseases increased dramatically during the latter part of the 20th century and millions of individuals are now affected. A number of scientists devised equipment to examine airborne biological particles during the 19th century, but aerobiological monitoring only became standardized during the 20th century. Airborne pollen are routinely monitored in many parts of the world, such as North America and Europe, and the first limited network has also been created for monitoring airborne allergen concentrations. Monitoring of the environment is often based on a combination of measurements and model results. Source-based models can increase our knowledge of airborne pollen because they can explain situations and processes that are almost impossible to understand using observations alone.
Assuntos
Pólen/imunologia , Rinite Alérgica Sazonal/patologia , Poluentes Atmosféricos/análise , Poluentes Atmosféricos/química , Alérgenos/química , Alérgenos/história , Alérgenos/imunologia , Monitoramento Ambiental , História do Século XVII , História do Século XIX , Humanos , Pólen/anatomia & histologia , Pólen/química , Rinite Alérgica Sazonal/história , Rinite Alérgica Sazonal/imunologiaRESUMO
BACKGROUND: Pollen of common ragweed (Ambrosia artemisiifolia) is a main cause of allergic diseases in Northern America. The weed has recently become spreading as a neophyte in Europe, while climate change may also affect the growth of the plant and additionally may also influence pollen allergenicity. To gain better insight in the molecular mechanisms in the development of ragweed pollen and its allergenic proteins under global change scenarios, we generated SuperSAGE libraries to identify differentially expressed transcripts. RESULTS: Ragweed plants were grown in a greenhouse under 380 ppm CO2 and under elevated level of CO2 (700 ppm). In addition, drought experiments under both CO2 concentrations were performed. The pollen viability was not altered under elevated CO2, whereas drought stress decreased its viability. Increased levels of individual flavonoid metabolites were found under elevated CO2 and/or drought. Total RNA was isolated from ragweed pollen, exposed to the four mentioned scenarios and four SuperSAGE libraries were constructed. The library dataset included 236,942 unique sequences, showing overlapping as well as clear differently expressed sequence tags (ESTs). The analysis targeted ESTs known in Ambrosia, as well as in pollen of other plants. Among the identified ESTs, those encoding allergenic ragweed proteins (Amb a) increased under elevated CO2 and drought stress. In addition, ESTs encoding allergenic proteins in other plants were also identified. CONCLUSIONS: The analysis of changes in the transcriptome of ragweed pollen upon CO2 and drought stress using SuperSAGE indicates that under global change scenarios the pollen transcriptome was altered, and impacts the allergenic potential of ragweed pollen.
Assuntos
Alérgenos/imunologia , Ambrosia/genética , Ambrosia/fisiologia , Dióxido de Carbono/farmacologia , Secas , Perfilação da Expressão Gênica , Pólen/imunologia , Estresse Fisiológico/genética , Ambrosia/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Cromatografia de Fase Reversa , Bases de Dados Genéticas , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Etiquetas de Sequências Expressas , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Biblioteca Gênica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Metabolismo Secundário/efeitos dos fármacos , Estresse Fisiológico/efeitos dos fármacos , Sobrevivência de Tecidos/efeitos dos fármacos , Sobrevivência de Tecidos/genética , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genéticaRESUMO
BACKGROUND: Evidence is compelling for a positive correlation between climate change, urbanisation and prevalence of allergic sensitisation and diseases. The reason for this association is not clear to date. Some data point to a pro-allergenic effect of anthropogenic factors on susceptible individuals. OBJECTIVES: To evaluate the impact of urbanisation and climate change on pollen allergenicity. METHODS: Catkins were sampled from birch trees from different sites across the greater area of Munich, pollen were isolated and an urbanisation index, NO2 and ozone exposure were determined. To estimate pollen allergenicity, allergen content and pollen-associated lipid mediators were measured in aqueous pollen extracts. Immune stimulatory and modulatory capacity of pollen was assessed by neutrophil migration assays and the potential of pollen to inhibit dendritic cell interleukin-12 response. In vivo allergenicity was assessed by skin prick tests. RESULTS: The study revealed ozone as a prominent environmental factor influencing the allergenicity of birch pollen. Enhanced allergenicity, as assessed in skin prick tests, was mirrored by enhanced allergen content. Beyond that, ozone induced changes in lipid composition and chemotactic and immune modulatory potential of the pollen. Higher ozone-exposed pollen was characterised by less immune modulatory but higher immune stimulatory potential. CONCLUSION: It is likely that future climate change along with increasing urbanisation will lead to rising ozone concentrations in the next decades. Our study indicates that ozone is a crucial factor leading to clinically relevant enhanced allergenicity of birch pollen. Thus, with increasing temperatures and increasing ozone levels, also symptoms of pollen allergic patients may increase further.
Assuntos
Poluentes Atmosféricos/análise , Alérgenos/imunologia , Betula/imunologia , Ozônio/análise , Pólen/imunologia , Dinoprostona/metabolismo , Ensaio de Imunoadsorção Enzimática , Humanos , Leucotrieno B4/metabolismo , Testes CutâneosRESUMO
Climate change and air pollution, including ozone is known to affect plants and might also influence the ragweed pollen, known to carry strong allergens. We compared the transcriptome of ragweed pollen produced under ambient and elevated ozone by 454-sequencing. An enzyme-linked immunosorbent assay (ELISA) was carried out for the major ragweed allergen Amb a 1. Pollen surface was examined by scanning electron microscopy and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), and phenolics were analysed by high-performance liquid chromatography. Elevated ozone had no influence on the pollen size, shape, surface structure or amount of phenolics. ATR-FTIR indicated increased pectin-like material in the exine. Transcriptomic analyses showed changes in expressed-sequence tags (ESTs), including allergens. However, ELISA indicated no significantly increased amounts of Amb a 1 under elevated ozone concentrations. The data highlight a direct influence of ozone on the exine components and transcript level of allergens. As the total protein amount of Amb a 1 was not altered, a direct correlation to an increased risk to human health could not be derived. Additional, the 454-sequencing contributes to the identification of stress-related transcripts in mature pollen that could be grouped into distinct gene ontology terms.
Assuntos
Ambrosia/imunologia , Antígenos de Plantas/imunologia , Ozônio/farmacologia , Proteínas de Plantas/imunologia , Pólen/imunologia , Antígenos de Plantas/análise , Mudança Climática , Ensaio de Imunoadsorção Enzimática , Ontologia Genética , Pólen/efeitos dos fármacos , Pólen/ultraestrutura , Estações do Ano , Espectroscopia de Infravermelho com Transformada de Fourier , Transcriptoma/efeitos dos fármacosRESUMO
The airway epithelium forms a physical, chemical and immunological barrier against inhaled environmental substances. In asthma, these barrier properties are thought to be abnormal. In this study, we analysed the effect of grass pollen on the physical and immunological barrier properties of differentiated human primary bronchial epithelial cells. Following exposure to Timothy grass (Phleum pratense) pollen extract, the integrity of the physical barrier was not impaired as monitored by measuring the transepithelial resistance and immunofluorescence staining of tight junction proteins. In contrast, pollen exposure affected the immunological barrier properties by modulating vectorial mediator release. CXC chemokine ligand (CXCL)8/interleukin (IL)-8 showed the greatest increase in response to pollen exposure with preferential release to the apical compartment. Inhibition of the extracellular signal-regulated kinase 1/2 and p38 mitogen-activated protein kinase pathways selectively blocked apical CXCL8/IL-8 release via a post-transcriptional mechanism. Apical release of CC chemokine ligand (CCL)20/macrophage inflammatory protein-3α, CCL22/monocyte-derived chemokine and tumour necrosis factor-α was significantly increased only in severe asthma cultures, while CCL11/eotaxin-1 and CXCL10/interferon-γ-induced protein-10 were reduced in nonasthmatic cultures. The bronchial epithelial barrier modulates polarised release of mediators in response to pollen without direct effects on its physical barrier properties. The differential response of cells from normal and asthmatic donors suggests the potential for the bronchial epithelium to promote immune dysfunction in asthma.
Assuntos
Asma/imunologia , Brônquios/patologia , Células Epiteliais/patologia , Extratos Vegetais/química , Pólen/química , Alérgenos/química , Asma/metabolismo , Broncoscopia , Células Cultivadas , Quimiocinas/imunologia , Humanos , Inflamação , Interleucina-8/imunologia , Ligantes , PoaceaeRESUMO
BACKGROUND: Nickel is a frequently detected cause of allergic contact dermatitis. Ingestion of nickel may lead to flares of nickel contact dermatitis. METHODS: We examined nickel excretion in the urine of 164 female patients with and without nickel contact dermatitis. The associations between age, atopic dermatitis, nickel contact dermatitis and nickel exposure through nutrition (e.g. dietary supplements) and by patch tests were investigated prospectively. Nickel was measured with atomic absorption spectrometry with two different standardized methods. RESULTS: A nickel detection limit of 0.2 µg/l was exceeded by all samples. The 95th percentiles of urine nickel concentration were 3.77 µg/l (age 18-30 years) and 3.98 µg/l (age 31-46 years). Bivariate analyses pointed to significantly increased nickel excretion with increasing age, ingestion of dietary supplements, drinking of stagnant tap water, and consumption of nickel-rich food. In the multivariate analysis, age and dietary supplements remained significant predictors of high nickel excretion. A non-significant increase in the median concentration of nickel was observed after the administration of conventional nickel patch tests. Patients with atopic eczema showed urine nickel concentrations similar to those in non-atopic controls. CONCLUSIONS: The 95th percentile of nickel excretion in our study population markedly exceeded the actual reference value of 3 µg/l. Age and consumption of dietary supplements are the most important predictors. The use of stagnant tap water and consumption of nickel-rich food contribute to the total load. These factors should be explicitly mentioned when allergic patients on a low-nickel diet are counselled. In contrast, existing nickel contact sensitization was not more frequent in subjects with higher nickel excretion. Nickel patch testing may cause transient minor systemic nickel exposure. The findings of this study extend our understanding and management of factors associated with nickel allergy.
Assuntos
Dermatite Alérgica de Contato/etiologia , Dieta/efeitos adversos , Exposição Ambiental/análise , Contaminação de Alimentos/análise , Níquel/efeitos adversos , Níquel/urina , Adulto , Bebidas/efeitos adversos , Dermatite Alérgica de Contato/diagnóstico , Exposição Ambiental/efeitos adversos , Feminino , Análise de Alimentos , Alemanha , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
The question what makes an allergen an allergen puzzled generations of researchers. Pollen grains of anemophilous plants are the most important allergen carriers in ambient air, and pollinosis is a highly prevalent multi-organ disease in civilized countries. In the past, research on the allergenicity of pollen has mainly focused on elucidating genetic predisposing factors and on defining certain structural characteristics of pollen derived allergens. Recently, studies extended to the analysis of non-allergenic, adjuvant mediators co-released from pollen. Besides active proteases and oxidases, extracts of pollen contain low molecular weight molecules like pollen-associated lipid mediators or adenosine exhibiting a potential to stimulate and modulate cultured human immune cells. This article reviews our current knowledge on non-allergenic, protein and non-protein compounds from pollen and their in vitro and in vivo effects on the allergic immune response. To ultimately judge the physiological relevance of these compounds, a systematic approach will be needed comparing their releasability, content and activity in different, allergenic and non-allergenic, pollen species. System biology such as proteome and metabolome analysis will be a useful future approach to better understand pollen biology.
Assuntos
Hipersensibilidade/imunologia , Pólen/imunologia , Animais , Quimiotaxia/imunologia , Humanos , Hipersensibilidade/genética , Estresse Oxidativo , Peptídeo Hidrolases/metabolismo , Pólen/química , Pólen/enzimologia , Biologia de SistemasRESUMO
Climate change with global warming is a physicometeorological fact that, among other aspects, will also affect human health. Apart from cardiovascular and infectious diseases, allergies seem to be at the forefront of the sequelae of climate change. By increasing temperature and concomitant increased CO(2) concentration, plant growth is affected in various ways leading to prolonged pollination periods in the northern hemisphere, as well as to the appearance of neophytes with allergenic properties, e.g. Ambrosia artemisiifolia (ragweed), in Central Europe. Because of the effects of environmental pollutants, which do not only act as irritants to skin and mucous membranes, allergen carriers such as pollen can be altered in the atmosphere and release allergens leading to allergen-containing aerosols in the ambient air. Pollen has been shown not only to be an allergen carrier, but also to release highly active lipid mediators (pollen-associated lipid mediators), which have proinflammatory and immunomodulating effects enhancing the initiation of allergy. Through the effects of climate change in the future, plant growth may be influenced in a way that more, new and altered pollens are produced, which may affect humans.
Assuntos
Mudança Climática , Hipersensibilidade/patologia , Alérgenos/imunologia , Dióxido de Carbono/química , Humanos , Hipersensibilidade/imunologia , Pólen/imunologiaRESUMO
OBJECTIVE AND METHODS: The crucial symptom of atopic eczema is itch. Acupuncture has been shown to exhibit a significant effect on experimental itch; however, studies focusing on clinical itch in atopic eczema and corresponding mechanisms are lacking. The study design was a unicenter, single-blinded (observer), prospective, randomized clinical pilot trial with an additional experimental part. In 10 patients with atopic eczema, we investigated the effect of acupuncture treatment (n = 5) compared to no treatment (n = 5) on itch intensity and in vitro basophil CD63 expression upon allergen stimulation (house dust mite and timothy grass pollen) in a pilot trial. RESULTS: Mean itch intensity in a visual analog scale was rated significantly lower in the acupuncture group (-25% ± 26% [day 15-day 0]; -24% ± 31% [day 33-day 0]) than in the control group (15% ± 6% [day 15-day 0]; 29% ± 9% [day 33-day 0]). From day 0 (before treatment) to day 15 (after 5 acupuncture treatments) as well as day 33 (after 10 acupuncture treatments), the acupuncture group showed less CD63 positive basophils than the control group regarding stimulation with house dust mite and grass pollen allergen at various concentrations (5 ng/mL, 1 ng/mL, 0.5 ng/mL, or 0.25 ng/mL). CONCLUSIONS: Our results show a reduction of itch intensity and of in vitro allergen-induced basophil activation in patients with atopic eczema after acupuncture treatment. Reducing basophil activation can be a further tool in investigating the mechanisms of action of acupuncture in immunoglobulin E-mediated allergy. Due to the limited number of patients included in our pilot trial, further studies are needed to strengthen the hypothesis.
Assuntos
Terapia por Acupuntura , Alérgenos/imunologia , Basófilos/imunologia , Dermatite Atópica/terapia , Prurido/terapia , Adulto , Animais , Dermatite Atópica/complicações , Dermatite Atópica/imunologia , Dermatophagoides pteronyssinus/imunologia , Feminino , Humanos , Masculino , Phleum/imunologia , Projetos Piloto , Prurido/etiologia , Prurido/imunologia , Índice de Gravidade de Doença , Método Simples-Cego , Tetraspanina 30/imunologia , Adulto JovemRESUMO
Anaphylaxis as the maximal variant of an acute systemic hypersensitivity reaction can involve several organ systems, particularly the skin, respiratory tract, gastrointestinal tract and the cardiovascular system. The severity of anaphylactic reaction is variable and can be classified into severity grades I-IV. Some reactions are fatal. Most frequent elicitors of anaphylaxis are foods in childhood, later insect stings and drugs. The phenomenon itself has been described in ancient medical literature, but was actually recognized and named at the beginning of the 20th century by Charles Richet and Paul Portier. In the course of experiments starting on the yacht of the Prince of Monaco and continued in the laboratory in Paris, they tried to immunize dogs with extracts of Physalia species in an attempt to develop an antitoxin to the venom of the Portuguese man-of-war. While Charles Richet believed that anaphylaxis was a 'lack of protection', it has become clear that an exaggerated immune reaction, especially involving immunoglobulin E antibodies, is the underlying pathomechanism in allergic anaphylaxis besides immune complex reactions. Non-immunologically mediated reactions leading to similar clinical symptomatology have been called 'anaphylactoid' or 'pseudo-allergic'--especially by Paul Kallos--and are now called 'non-immune anaphylaxis' according to a consensus of the World Allergy Organization (WAO). The distinction of different pathophysiological processes is important since non-immune anaphylaxis cannot be detected by skin test or in vitro allergy diagnostic procedures. History and provocation tests are crucial. The intensity of the reaction is not only influenced by the degree of sensitization but also by concomitant other factors as age, simultaneous exposure to other allergens, underlying infection, physical exercise or psychological stress or concomitant medication (e.g. beta-blockers, NSAIDs); this phenomenon has been called augmentation or summation anaphylaxis.
Assuntos
Anafilaxia/história , Hipersensibilidade a Drogas/história , Hipersensibilidade Alimentar/história , Alérgenos/imunologia , Anafilaxia/classificação , Anafilaxia/diagnóstico , Anafilaxia/imunologia , Animais , Criança , Diagnóstico Diferencial , Cães , Hipersensibilidade a Drogas/classificação , Hipersensibilidade a Drogas/diagnóstico , Hipersensibilidade a Drogas/etiologia , Hipersensibilidade a Drogas/imunologia , Hipersensibilidade Alimentar/classificação , Hipersensibilidade Alimentar/diagnóstico , Hipersensibilidade Alimentar/imunologia , História do Século XX , História Antiga , História Medieval , Humanos , Imunização , Imunoglobulina E/imunologia , ParisRESUMO
A subgroup of patients with atopic eczema develops acute eczematous reactions to type I allergy-inducing agents such as pollen that clinically resemble type IV allergies induced by haptens like metal ions. To clarify the underlying immunologic mechanisms, this study was designed to map the inflammatory in situ topoproteome of eczematous responses to grass/birch pollen and nickel by using atopy patch test (APT) and nickel patch test (NPT) as an appropriate clinical model, respectively. Biopsies from NPT (n = 6) and APT (n = 6) with positive reactions at 72 h were analysed by multiple epitope ligand cartography (MELC), which enabled to investigate coexpression of 49 different epitopes immunohistochemically in a single given tissue section. Colocalisation of IgE and FcepsilonRI was investigated by confocal microscopy. Compared with APT responses, NPT reactions were dominated by cytotoxic TIA-1 + and CD8 + T cells. In contrast, the immune response in APT reactions appeared more pleiotrope - as detected by colocalisation analysis. Multiple combinatorial molecular phenotype (CMP) motifs containing naive, early maturation and memory T cell (CD45RA, CD7, CD44, CD45R0), and general activation markers (CLA, HLA-DR, CD13, CD29, CD58, CD71, CD138) were significantly higher expressed in APT when compared with NPT reactions. APT response was confirmed to be accompanied by IgE bound to FcepsilonRI. In summary, our results demonstrate that the NPT reaction is clearly dominated by cytotoxic events, while the APT reaction to pollen grains is more heterogeneous and elicits a combined humoral and cellular immune reaction.
Assuntos
Eczema/imunologia , Imunidade Celular/imunologia , Imunidade Humoral/imunologia , Níquel/imunologia , Testes do Emplastro , Pólen/imunologia , Proteoma/imunologia , Antígenos CD/metabolismo , Antígenos de Diferenciação de Linfócitos T/metabolismo , Biópsia , Moléculas de Adesão Celular/metabolismo , Contagem de Células , Movimento Celular/imunologia , Citotoxicidade Imunológica/imunologia , Células Dendríticas/citologia , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Dermatite Alérgica de Contato/imunologia , Dermatite Atópica/imunologia , Antígenos HLA-D/metabolismo , Humanos , Imunoglobulina E/metabolismo , Leucócitos/citologia , Leucócitos/imunologia , Leucócitos/metabolismo , Ativação Linfocitária/imunologia , Proteínas de Ligação a Poli(A)/metabolismo , Proteoma/metabolismo , Proteômica/métodos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Receptores de IgE/metabolismo , Pele/imunologia , Pele/metabolismo , Pele/patologia , Antígeno-1 Intracelular de Células T , Linfócitos T/citologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Linfócitos T Citotóxicos/citologia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/metabolismoRESUMO
In a humid milieu such as mucosal surfaces, pollen grains do not only release allergens but also proinflammatory and immunomodulatory lipids, termed pollen-associated lipid mediators. Among these, the E(1)-phytoprostanes (PPE(1)) were identified to modulate dendritic cell (DC) function: PPE(1) inhibit the DC's capacity to produce IL-12 and enhance DC mediated T(H)2 polarization of naive T cells. The mechanism(s) by which PPE(1) act on DC remained elusive. We thus analyzed candidate signaling elements and their role in PPE(1)-mediated regulation of DC function. Aqueous birch pollen extracts induced a marked cAMP response in DC that could be blocked partially by EP2 and EP4 antagonists. In contrast, PPE(1) hardly induced cAMP and the inhibitory effect on IL-12 production was mostly independent of EP2 and EP4. Instead, PPE(1) inhibited the LPS-induced production of IL-12 p70 by a mechanism involving the nuclear receptor PPAR-gamma. Finally, PPE(1) efficiently blocked NF-kappaB signaling in DCs by inhibiting IkappaB-alpha degradation, translocation of p65 to the nucleus, and binding to its target DNA elements. We conclude that pollen-derived PPE(1) modulate DC function via PPAR-gamma dependent pathways that lead to inhibition of NFkappaB activation and result in reduced DC IL-12 production and consecutive T(H)2 polarization.
Assuntos
Ciclopentanos/imunologia , Células Dendríticas/imunologia , Ácidos Graxos Insaturados/imunologia , Interleucina-12/biossíntese , NF-kappa B/antagonistas & inibidores , PPAR gama/metabolismo , Pólen/imunologia , Transdução de Sinais/imunologia , Adolescente , Adulto , Betula/imunologia , Células Cultivadas , AMP Cíclico , Humanos , Fatores Imunológicos , Pessoa de Meia-Idade , NF-kappa B/metabolismo , Receptores de Prostaglandina E/metabolismo , Receptores de Prostaglandina E Subtipo EP2 , Receptores de Prostaglandina E Subtipo EP4 , Células Th2 , Adulto JovemRESUMO
Epithelial cells of both the respiratory tract and the skin form a tight barrier against environmental harm. They represent the site of first contact for airborne allergen carriers. Consequently, in this study, we analyzed the uptake of grass pollen allergens by epithelial cells: Phl p 1 was selected as a glycosylated allergen containing disulfide bridges whereas Phl p 6 lacks post-translational modifications. Allergen uptake by the respiratory epithelial cell line A549 reached a plateau at 2 hours, and both allergens were localized intracellularly in non-acidic vesicles. In addition, in A549 cells allergens were exocytosed, suggesting a transcytosis mechanism in the passage of allergens over the respiratory epithelial barrier. In contrast, allergens were predominately localized in lysosomes in keratinocytes, and allergen uptake did not reach a plateau up to 24 hours. Notably, keratinocytes from atopic patients showed a significantly increased uptake of Phl p 1 as compared with healthy donors. Preincubation of epithelial cells with IL-4 and/or IFN-gamma to simulate inflammatory status led to an increased allergen uptake only in keratinocytes. This higher engulfment of allergens by inflammatory-type keratinocytes suggests a higher susceptibility of inflamed skin for the uptake of allergens and consequently a potentially higher risk for sensitization under natural exposure conditions, such as chronic atopic eczema.
Assuntos
Alérgenos/metabolismo , Brônquios/metabolismo , Células Epiteliais/metabolismo , Queratinócitos/metabolismo , Poaceae/imunologia , Pólen/imunologia , Células Cultivadas , Exocitose , Humanos , Interferon gama/farmacologia , Interleucina-4/farmacologia , Fatores de TempoRESUMO
INTRODUCTION: Eczematous reactions to type I allergy-inducing antigens are documented in a subgroup of patients with atopic eczema. Yet, the underlying immunological mechanisms are not well understood. MATERIAL AND METHODS: To delineate the effect of native pollen grains on human skin of healthy and atopic individuals we performed patch tests (atopy patch test with native pollen grains, PPT). Nickel patch tests (NPT) served as an established model of contact dermatitis. Skin site biopsies were taken 6-96 h after allergen application and investigated immunohistochemically. RESULTS: Histology of positive patch tests showed an influx of mononuclear cells (predominantly CD4+, CD25+, CD45RO+). This influx was detected earlier in the PPT reaction than in the immune response to nickel. A biphasic cytokine response could be detected in the PPT: IL-5 dominated in the early, IFN-gamma in the late phase. The NPT was continuously dominated by IFN-gamma. Dendritic cell subpopulations imitated the earlier kinetics of the mononuclear infiltrate. DISCUSSION: Thus, pollen grains induce eczematous reactions in susceptible individuals. This reaction appears clinically and immunohistochemically similar to the contact hypersensitivity reaction to nickel but follows a faster kinetic and a biphasic course: Th2 and IgE in the early (24 h) and Th1 predominance in the late (96 h) phase.