Release of Bet v 1 from Birch Pollen 1 from 5 European 2 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 grain 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 l/min at sites in France, United Kingdom, Germany, Italy and Finland. Allergen concentrations in ambient air were sampled at 800l/min with a Chemvol high-volume cascade impactor equipped with stages PM>10μm, 10 μm>PM>2.5μm, and in Germany also 2.5 μm>PM>0.12μm. 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 FcεR1-humanized rat basophil cell line passively sensitized with serum of a birch pollen lmptomatic 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>10μm fraction at all stations. Bet v 1 isoforms pattern did not varied 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.

Variation of the Group 5 Grass Pollen Allergen Content of Airborne Pollen in Relation to Geographical Location and Time in Season

Allergies to grass pollen are the number one cause of outdoor hay fever. The human immune system reacts with symptoms to allergens 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 was determined with a Phl p 5 specific ELISA in two fractions of ambient air: Particulate Matter (PM) >10μm and 10μm>PM>2.5μm. Mediator release by ambient air was determined in FcεR1-humanized basophils. Origin of pollen was modeled and condensed to pollen potency maps. Results: On average grass pollen released 2.3 pg Phl p 5/pollen. Allergen release per pollen (potency) varied substantially, ranging from 0 to 9 pg Phl p 5/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/m3 (r2=0.80, p<0.001) than with pollen/m3 (r2=0.61, p<0.001). In addition, pollen released different amounts of allergen in the nonpollen 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 on top of variations in pollen counts. Molecular aerobiology, i.e. determining allergen in ambient air, may be a valuable addition to pollen counting.