Particle size distribution of the major Alternaria alternata allergen, Alt a 1, derived from airborne spores and subspore fragments.

Fungal fragments are abundant immunoreactive bioaerosols that may outnumber the concentrations of intact spores in the air. To investigate the importance of Alternaria fragments as sources of allergens compared to Alternaria spores, we determined the levels of Alternaria spores and Alt a 1 (the major allergen in Alternaria alternata spores) collected on filters within three fractions of particulate matter (PM) of different aerodynamic diameter: (1) PM>10, (diameter>10 µm); (2) PM2.5-10 (2.5-10µm); (3) PM2.5 (0.12-2.5 µm). The airborne particles were collected using a three stage high-volume ChemVol cascade impactor during the Alternaria sporulation season in Poznan, Poland (30 d between 6 July and 22 September 2016). The quantification of Alt a 1 was performed using the enzyme-linked immunosorbent assay. High concentrations of Alt a 1 were recorded during warm and dry d characterized by high sunshine duration, lack of clouds and high dew point values. Atmospheric concentrations of Alternaria spores correlated significantly (r = 0.930, p < 0.001) with Alt a 1 levels. The highest Alt a 1 was recorded in PM2.5-10 (66.8 % of total Alt a 1), while the lowest in PM2.5 (<1.0 %). Significantly more Alt a 1 per spore (>30 %) was observed in PM2.5-10 than in PM>10. This Alt a 1 excess may be derived from sources other than spores, e.g. hyphal fragments. Overall, in outdoor air the major source of Alt a 1 are intact Alternaria spores, but the impact of other fungal fragments (hyphal parts, broken spores, conidiophores) cannot be neglected, as they may increase the total atmospheric Alt a 1 concentration.

Atmospheric exposure to the major Artemisia pollen allergen (Art v 1): Seasonality, impact of weather, and clinical implications.

Artemisia pollen grains are important aeroallergens worldwide. The amount of allergenic proteins produced by pollen, or pollen allergenicity, is regulated by both genes and the environment. As a result, even closely related plant taxa may release pollen with distinctly different allergen contents. Here, we determined the variability in atmospheric exposure to the major Artemisia pollen allergen, Art v 1, during the pollination seasons of two common species, i.e., A. vulgaris (early flowering species) and A. campestris (late flowering species), in Poznan, Poland (2013-2015). Artemisia pollen grains were collected using Hirst-type volumetric trap, while Art v 1 was collected by a two-stage cascade impactor (PM10 and PM>10 air fractions) and quantified by immunoenzymatic analysis. The results showed that daily Art v 1 levels correlated significantly with mean daily concentrations of Artemisia pollen (from r = 0.426 to r = 0.949, depending on air fraction and peak of the season). Significant differences were observed between 1) the median pollen allergenicity in different seasons (from 2.5 to 4.7 pg Art v 1/pollen) and 2) the median pollen allergenicity in different peak periods of the season (from 1.8 to 6.7 pg Art v 1/pollen). During the late peak (flowering of A. campestris), the median pollen allergenicity was significantly higher (on average by 63%, p < 0.05) than that during A. vulgaris flowering. The highest mean seasonal pollen allergenicity was observed during the wettest season, while the lowest was observed during the driest season (from July-August). In summary, our study showed distinct differences in Artemisia pollen allergenicity, that were not only related to daily and seasonal variability, which may exceed 800% and 80%, respectively but also noticeable when two common Artemisia species were compared. Therefore, we argue that variability in pollen allergenicity (both seasonal and species-specific) should be considered in future studies assessing pollen exposure.

Temporal variability in the allergenicity of airborne Alternaria spores.

The concentration of fungal spores in the air is traditionally considered as a proxy of allergen exposure. However, in vitro experiments have shown that the allergenicity of Alternaria spores varies depending on ecophysiological and developmental factors. Despite the potential clinical significance of these findings, it has never been verified in outdoor environments. This study, therefore, aims to investigate variability in the amount of the major allergen (Alt a 1) released from Alternaria spores in outdoor air. During the 3-year monitoring study (2014-2016), the median seasonal allergenicity of Alternaria spores exceeded 8.6 × 10-3 pg Alt a 1/spore. The most allergenic spores were collected during the driest and the most polluted season (with respect to seasonal concentrations of ozone, sulphur dioxide, and particulate matter). Within the season, daily spore allergenicity ranged from 2.4 to 34.7 × 10-3 pg Alt a 1/spore (5th-95th percentile). No repeatable effects of weather and pollution on short-term variations in Alternaria spore allergenicity were found. However, during the episodes when high-potency spores were recorded, the air masses arrived from eastern directions. Contrary, the spores with the lowest allergenicity were related to western winds. This suggests that factors such as source area (habitat types) and species diversity could be responsible for the varying exposure to Alternaria allergens. Our findings show that high and low-potency spores are recorded in the air; therefore, the airborne concentrations of fungal spores alone may not be sufficient to provide allergy sufferers and healthcare professionals with information about allergen exposure.