THE HIALINE PROJECT: ALLERGEN RELEASE FROM POLLEN ACROSS 10 EUROPEAN COUNTRIES

Exposure to allergens is one of severa1 factors determining sensitization and allergic symptoms in individuals. Exposure to aeroallergens from pollen is assessed by counting allergenic pollen in ambient air. However, proof is lacking that pollen count is representative for allergen exposure. We therefore monitored simultaneously birch, grass and olive pollen counts and their corresponding major pollen allergens Bet v 1, Phl p 5 and Ole e 1 across Europe. Already at one location in Europe in Munich, Germany, it has been found that the same amount of pollen from different years, different trees and even different days released up to lO-fold different amounts of Bet v 1. Thus exposure to allergen is poorly monitored by only monitoring pollen countl-2. Monitoring the allergen itself in ambient air might be an improvement in allergen exposure assessment. The objective of the HIALINE-project is to evaluate if these effects found in Munich, Germany are also measurable over a bigger geographic area like Europe, and at the same time implement an outdoor allergen early warning network, in addition to the pollen forecasts. Climatic factors that influence allergen exposure will be extracted and will be used to calculate the effect of climate change on local airborne allergen exposure. The major allergens from the top 3 airborne allergens in Europe (grasses, birch and olive) are sampled with a cascade impactor, extracted and analyzed by allergen specific ELISA 's. Pollen counts are measured by standard pollen traps and correlated with the weather data. Allergen forecast will be calculated by incorporating the SILAM chemical transport model and compared with the observations of HIALINE aiming at a comprehensive parameterization of the allergen release and transport. Expected outcomes are the implementation of a network of European outdoor allergen measurements to better predict allergic symptoms. Also the climatic factors that govern allergen exposure in outdoor air will be established. These can be used to calculate the effect of climate change on the health effects of airborne allergens The research leading to these results has received funding from the Executive Agency for Health and Consumers under grant agreement No 2008 11 07

The European project HIALINE (Health Impacts of Airborne Allergen Information Network): results of pollen and allergen of Betula monitoring in Parma (2009)

Introduction. Exposure to allergens is pivotal in determining sensitization and allergic symptoms in individuals. Pollen grain counts in ambient air have traditionally been assessed to estimate airborne allergen exposure. However, the exact allergen content in ambient air is unknown. HIALINE therefore monitored atmospheric concentrations of Betula, Poaceae and Olea pollen grains and matched their major allergens Bet v1, Phl p5 and Ole e1 across Europe. Monitoring the allergens themselves together with pollen in ambient air might be an improvement in allergen exposure assessment. New knowledge through the use of new experimental approaches in the field of aerobiological monitoring will enable better in the prevention and clinical management of pollinosis. In order to disseminate the knowledge of the project we present the results of first year of birch pollen grains and the matched major pollen allergen Bet v1 monitored in Parma (UNIPR), Italy with a short reference to the results obtained by the other participants and about developing models of dissemination and forecasts of pollen and allergens. Materials and methods. The pollen was sampled by a Hirst pollen trap. Allergens was collected with a CHEMVOL® high-volume cascade impactor, extracted from pollen and quantified by ELISA. Antibodies for analysis of Bet v1 are delivered by the industrial partner in this project. Quality control has been carried out for the monitoring pollen activities and allergen concentrations. Results. The project has highlighted that it is possible also to measure pollen allergen in ambient air in different European areas. The results obtained from the center of Parma and other European partners have highlighted the different allergenic powers of pollen of Betula, in different geographical areas. Moreover, daily in each area the allergenic power of pollen grains was very variable. Discussion. HIALINE has been a very important project to understand the mechanisms of sensitization, clinical management of pollinosis and to improve immunotherapy towards a tailored immunotherapy. The results of the project will help medical doctors, authorities and patients, to better manage the different aspects related to pollinosis

Airborne olive pollen counts are not representative of exposure to the major olive allergen Ole e 1

Pollen is routinely monitored, but it is unknown whether pollen counts represent allergen exposure. We therefore simultaneously determined olive pollen and Ole e 1 in ambient air in C ordoba, Spain, and Evora, Portugal, using Hirst-type traps for pollen and high-volume cascade impactors for allergen. Pollen from different days released 12-fold different amounts of Ole e 1 per pollen (both locations P < 0.001). Average allergen release from pollen (pollen potency) was much higher in C ordoba (3.9 pg Ole e 1/pollen) than in Evora (0.8 pg Ole e 1/pollen, P = 0.004). Indeed, yearly olive pollen counts in C ordoba were 2.4 times higher than in Evora, but Ole e 1 concentrations were 7.6 times higher. When modeling the origin of the pollen, >40% of Ole e 1 exposure in Evora was explained by high-potency pollen originating from the south of Spain. Thus, olive pollen can vary substantially in allergen release, even though they are morphologically identical

Outdoor airborne allergens: Characterization, behavior and monitoring in Europe

Aeroallergens or inhalant allergens, are proteins dispersed through the air and have the potential to induce allergic conditions such as rhinitis, conjunctivitis, and asthma. Outdoor aeroallergens are found predominantly in pollen grains and fungal spores, which are allergen carriers. Aeroallergens from pollen and fungi have seasonal emission patterns that correlate with plant pollination and fungal sporulation and are strongly associated with atmospheric weather conditions. They are released when allergen carriers come in contact with the respiratory system, e.g. the nasal mucosa. In addition, due to the rupture of allergen carriers, airborne allergen molecules may be released directly into the air in the form of micronic and submicronic particles (cytoplasmic debris, cell wall fragments, droplets etc.) or adhered onto other airborne particulate matter. Therefore, aeroallergen detection strategies must consider, in addition to the allergen carriers, the allergen molecules themselves. This review article aims to present the current knowledge on inhalant allergens in the outdoor environment, their structure, localization, and factors affecting their production, transformation, release or degradation. In addition, methods for collecting and quantifying aeroallergens are listed and thoroughly discussed. Finally, the knowledge gaps, challenges and implications associated with aeroallergen analysis are describe

Mesoscale Atmospheric Transport of Ragweed Pollen Allergens from Infected to Uninfected Areas

Allergenic ragweed (Ambrosia spp.) pollen grains, after being released from anthers, can be dispersed by air masses far from their source. However, the action of air temperature,humidity and solar radiation on pollen grains in the atmosphere could impact on the ability of long distance transported (LDT) pollen to maintain allergenic potency. Here, we report that the major allergen of Ambrosia artemisiifolia pollen (Amb a 1) collected in ambient air during episodes of LDT still have immunoreactive properties. The amount of Amb a 1 found in LDT ragweed pollen grains was not constant and varied between episodes. In addition to allergens in pollen sized particles, we detected reactive Amb a 1 in subpollen sized respirable particles. These findings suggest that ragweed pollen grains have the potential to cause allergic reactions, not only in the heavily infested areas but, due to LDT episodes, also in the regions unaffected by ragweed populations

Release of Bet v 1 from birch pollen from 5 European countries. Results from the HIALINE study

Exposure to allergens is pivotal in determining sensitization and allergic symptoms in individuals. Pollen grain counts in ambient air have traditionally been assessed to estimate airborne allergen exposure. However, the exact allergen content of ambient air is unknown. We therefore monitored atmospheric concentrations of birch pollen grains and the matched major birch pollen allergen Bet v 1 simultaneously across Europe within the EU-funded project HIALINE (Health Impacts of Airborne Allergen Information Network). Pollen count was assessed with Hirst type pollen traps at 10 I min(-1) at sites in France, United Kingdom, Germany, Italy and Finland. Allergen concentrations in ambient air were sampled at 800 I min(-1) with a Chemvol (R) high-volume cascade impactor equipped with stages PM > 10 mu m, 10 mu m > PM > 2.5 mu m, and in Germany also 2.5 mu m > PM > 0.12 mu m. The major birch pollen allergen Bet v 1 was determined with an allergen specific ELISA. Bet v 1 isoform patterns were analyzed by 2D-SDS-PAGE blots and mass spectrometric identification. Basophil activation was tested in an FC epsilon R1-humanized rat basophil cell line passively sensitized with serum of a birch pollen symptomatic patient. Compared to 10 previous years, 2009 was a representative birch pollen season for all stations. About 90% of the allergen was found in the PM > 10 mu m fraction at all stations. Bet v 1 isoforms pattern did not vary substantially neither during ripening of pollen nor between different geographical locations. The average European allergen release from birch pollen was 3.2 pg Bet v 1/pollen and did not vary much between the European countries. However, in all countries a >10-fold difference in daily allergen release per pollen was measured which could be explained by long-range transport of pollen with a deviating allergen release. Basophil activation by ambient air extracts correlated better with airborne allergen than with pollen concentration. Although Bet v 1 is a mixture of different isoforms, its fingerprint is constant across Europe. Bet v 1 was also exclusively linked to pollen. Pollen from different days varied >10-fold in allergen release. Thus exposure to allergen is inaccurately monitored by only monitoring birch pollen grains. Indeed, a humanized basophil activation test correlated much better with allergen concentrations in ambient air than with pollen count. Monitoring the allergens themselves together with pollen in ambient air might be an improvement in allergen exposure assessment.European CommissionChristine Kühne - Center for Allergy Research and Educatio