Pollen allergens, widely present in the atmosphere, are the main cause of seasonal respiratory diseases that affect millions of people worldwide. Although previous studies have reported that nitrogen dioxide (NO₂) and ozone (O₃) promote pollen allergy, the specific biological processes and underlying mechanisms remain less understood. In this study, Platanus pollen grains were exposed to gaseous pollutants (NO₂ and O₃). We employed environmental electron microscopy, flow cytometry, western blot assay, enzyme-linked immunoassay, ultraviolet absorption spectrometry, circular dichroism, and protein mass spectrometry to characterise the subpollen particles (SPPs) released from pollen grains. Furthermore, we determined the immunogenicity and pathogenicity induced by Platanus pollen allergen a 3 (Pla a 3). Our results demonstrated that NO₂ and O₃ could damage the pollen cell membranes in SPPs and increase the amount of Pla a 3 allergen released into the atmosphere. Additionally, NO₂ and O₃ altered the structure of Pla a3 protein through nitrification and oxidation, which not only enhanced the immunogenicity of allergens but also increased the stability of the protein. In vivo analysis using an animal model indicated that NO₂ and O₃ greatly aggravated pollen-induced pneumonia. Thus, our study provides guidance for the prevention of pollen allergic diseases.

Ozone (O₃) and nitrogen dioxide (NO₂) are thought to play primary roles in aggravating air pollution-induced health problems. However, the effects of joint O₃/NO₂ on the allergenicity of pollen allergens are unclear. Humulus japonicus pollen allergen 1 (Hum j1) is a profilin protein that causes widespread pollinosis in eastern Asia. In order to study the effects of combined O₃/NO₂ on the allergenicity of Hum j1, tandem six-histidine peptide tag (His6)-fused recombinant Hum j1 (rHum j1) was expressed in a prokaryotic system and purified through His6 affinity chromatography. The purified rHum j1 was used to immunize SD rats. Rat sera with high titers of IgG and IgE antibodies against rHum j1 were used for allergenicity quantification. The rHum j1 was exposed to O₃/NO₂, and changes in allergenicity of the exposed rHum j1 were assayed using the immunized rat antibodies. Tandem LC-MS/LC (liquid chromatography-mass spectrometer/liquid chromatography spectrometer) chromatography and UV and circular dichroism (CD) spectroscopy were used to study the structural changes in rHum j1. Our data demonstrated that a novel disulfide bond between the sulfhydryl groups of two neighboring cysteine molecules was formed after the rHum j1 exposure to joint O₃/NO₂, and therefore IgE-binding affinity was increased and the allergenicity was reinforced. Our results provided clues to elucidate the mechanism behind air pollution-induced increase in pollinosis prevalence.

Common ragweed (Ambrosia artemisiifolia L.) is a highly allergenic annual ruderal plant and native to Northern America, but now also spreading across Europe. Air pollution and climate change will not only affect plant growth, pollen production and duration of the whole pollen season, but also the amount of allergenic encoding transcripts and proteins of the pollen. The objective of this study was to get a better understanding of transcriptional changes in ragweed pollen upon NO2 and O3 fumigation. This will also contribute to a systems biology approach to understand the reaction of the allergenic pollen to air pollution and climate change. Ragweed plants were grown in climate chambers under controlled conditions and fumigated with enhanced levels of NO2 and O3. Illumina sequencing and de novo assembly revealed significant differentially expressed transcripts, belonging to different gene ontology (GO) terms that were grouped into biological process and molecular function. Transcript levels of the known Amb a ragweed encoding allergens were clearly up-regulated under elevated NO2, whereas the amount of allergen encoding transcripts was more variable under elevated O3 conditions. Moreover transcripts encoding allergen known from other plants could be identified. The transcriptional changes in ragweed pollen upon elevated NO2 fumigation indicates that air pollution will alter the transcriptome of the pollen. The changed levels of allergenic encoding transcripts may have an influence on the total allergenic potential of ragweed pollen.

Air pollution is a widespread health problem associated with respiratory symptoms. Continuous exposure monitoring was performed to estimate alveolar and tracheobronchial dose, measured as deposited surface area, for 103 children and to evaluate the long-term effects of exposure to airborne particles through spirometry, skin prick tests and measurement of exhaled nitric oxide (eNO). The mean daily alveolar deposited surface area dose received by children was 1.35 × 103 mm2. The lowest and highest particle number concentrations were found during sleeping and eating time. A significant negative association was found between changes in pulmonary function tests and individual dose estimates. Significant differences were found for asthmatics, children with allergic rhinitis and sensitive to allergens compared to healthy subjects for eNO. Variation is a child's activity over time appeared to have a strong impact on respiratory outcomes, which indicates that personal monitoring is vital for assessing the expected health effects of exposure to particles.

Asthma is currently defined as a chronic inflammatory disease of the airway. Several evidence indicate that vehicle emissions in cities is correlated with the allergic respiratory diseases. In the present study, we evaluated in the A549 cells the production and release of IL-4, IL-5 and IL-13 after treatment with sub-micron PM₁.₀ particles (PM₁.₀), Parietaria officinalis (ALL), and PM₁.₀ + ALL together. Our data demonstrated that PM₁.₀ + ALL together exhibited the greatest capacity to induce A549 cells to enhance the expression of IL-4 and IL-5 compared with the only PM₁.₀ or ALL treatment. Interestingly, IL-13 that is necessary for allergen-induced airway hyper responsiveness, is increased in cells treated with PM₁.₀ + ALL together, but is higher expressed when the cells are treated only with the allergen. Our data support the hypothesis that the urban environment damage the acinar lung units and activates cells of the immune system.

The prevalence of allergic diseases in Australian children is high, but few studies have assessed the potential role of outdoor air pollution in allergic sensitization. We investigated the association between outdoor air pollution and the prevalence of aeroallergen sensitization in a national cross-sectional study of Australian children aged 7–11 years. Children were recruited from 55 participating schools in 12 Australian cities during 2007–2008. Parents completed a detailed (70-item) questionnaire. Outdoor nitrogen dioxide (NO₂), as a proxy for exposure to traffic-related emissions, was estimated using measurements from regulatory monitors near each school and a national land-use regression (LUR) model. Three averaging periods were assessed, using information on duration of residence at the address, including lifetime, previous (lifetime, excluding the last year), and recent (the last year only). The LUR model was used as an additional source of recent exposure estimates at school and home addresses. Skin prick tests (SPTs) were performed to measure sensitization to eight common aeroallergens. Multilevel logistic regression estimated the association between NO₂ and sensitization (by individual allergens, indoor and outdoor allergens, and all allergens combined), after adjustment for individual- and area-level covariates. In total, 2226 children had a completed questionnaire and SPT. The prevalence of sensitization to any allergen was 44.4%. Sensitization to house dust mites (HDMs) was the most common (36.1%), while sensitization to Aspergillus was the least common (3.4%). Measured mean (±s.d.) NO₂ exposure was between 9 (±2.9) ppb and 9.5 (±3.2) ppb, depending on the averaging period. An IQR (4 ppb) increase in measured previous NO₂ exposure was associated with greater odds of sensitization to HDMs (OR: 1.21, 95% CI: 1.01–1.43, P = 0.035). We found evidence of an association between relatively low outdoor NO₂ concentrations and sensitization to HDMs, but not other aeroallergens, in Australian children aged 7–11 years.

Air pollution is frequently proposed as a cause of the increased incidence of allergy in industrialised countries. We investigated the impact of ozone (O₃) on reactive oxygen species (ROS) and allergen content of ragweed pollen (Ambrosia artemisiifolia). Pollen was exposed to acute O₃ fumigation, with analysis of pollen viability, ROS and nitric oxide (NO) content, activity of nicotinamide adenine dinucleotide phosphate (NAD[P]H) oxidase, and expression of major allergens. There was decreased pollen viability after O₃ fumigation, which indicates damage to the pollen membrane system, although the ROS and NO contents were not changed or were only slightly induced, respectively. Ozone exposure induced a significant enhancement of the ROS-generating enzyme NAD(P)H oxidase. The expression of the allergen Amb a 1 was not affected by O₃, determined from the mRNA levels of the major allergens. We conclude that O₃ can increase ragweed pollen allergenicity through stimulation of ROS-generating NAD(P)H oxidase.

Olive pollen is one of the main causes of allergic disease in the Mediterranean area. Ten different proteins with allergenic activity have been described in olive pollen, with major allergen Ole e 1. Olea europaea L. may cause allergenic effects of different severity depending on the Ole e 1 content of cultivars. In this paper, we aimed to assess the heterogeneity of two olive cultivars concerning concentrations of the major allergen Ole e 1 during a period of 2 years. Pollens from two most common olive cultivars, known as “Gemlik” and “Celebi,” were analyzed on regular basis. Ole e 1 amounts were measured by double-sandwich enzyme-linked immunosorbent assay (ELISA). The results were expressed as μg of Ole e 1 per μg of total freeze-dried extract. Comparisons of Ole e 1 levels were made both between individual trees and between cultivars. It was analyzed the influence of some meteorological parameters on pollen counts/allergenic content on a local scale, for 2 years. Pollen sampling was carried out continuously for 2 years, using a Hirst-type volumetric trap. “Gemlik” had the higher value (mean ± standard deviation) of Ole e 1 content (2.44 ±0.70 and 1.87 ±1.03 μg/μg, respectively) when compared to “Celebi” (2.16 ±0.86 and 0.20 ±0.30 μg/μg, respectively) in the years 2013 and 2015. In our research, daily variations were observed in pollen samples of two olive cultivars and even different trees of the same cultivar. Furthermore, during certain sampling days, discrepancies between airborne pollen counts and Ole e 1 concentrations were detected for both cultivars. It was found that meteorological changes, especially temperature and precipitation fluctuations, could affect airborne pollen and Ole e 1 allergen levels in the atmosphere. Therefore, pollen samples of different O. europaea cultivars demonstrated great differences in Ole e 1 content. We believe that these findings were a result of alternate bearing behavior modulated by meteorological factors.

The aim of this study was to compare airborne levels of Phl p 1 and Phl p 5, with Poaceae pollen concentrations inside and outside of the pollen season, and to evaluate their association with symptoms in grass allergic patients and the influence of climate and pollution. The Hirst and the Burkard Cyclone samplers were used for pollen and allergen quantification, respectively. The sampling period ran from 23 March 2009 to 27 July 2010. Twenty-three patients with seasonal allergic asthma and rhinitis used an electronic symptom card. The aerosol was extracted and quantified for Phl p 1 and Phl p 5 content. Descriptive statistics, non-parametric paired contrast of Wilcoxon, Spearman’s correlations, and a categorical principal component analysis (CatPCA) were carried out. Significant variations in pollen, aeroallergen levels, pollen allergen potency, and symptoms score were observed in this study. Phl p 5 pollen allergen potency was higher at the beginning of the 2010 grass pollen season. Presence of Phl p 1 outside the pollen season with positive O₃ correlation was clinically relevant. 45.5% of the variance was explained by two dimensions in the CatPCA analysis, showing the symptom relationships dissociated in two dimensions. In the first one, the more important relationship was with grass pollen grains concentration and Phl p 5 and to a lesser extent with Phl p 1 and levels of NO₂ and O₃, and in the second dimension, symptoms were associated with humidity and SO₂. Clinically relevant out-season Phl p 1 was found with a positive O₃ correlation. The effect of climate and pollution may have contributed to the higher seasonal allergic rhinitis symptom score recorded in 2009.

Birch (Betula pendula) pollen causes inhalant allergy in about 20% of human population in Europe, most of which is sensitive to the main birch allergen, Bet v1. The aim of the study was to find out (i) whether and how the analysed birch individuals differ in regard to composition of individual subunits of pollen proteins and to protein content in these subunits; (ii) whether the level of particulate matter relates to concentration of Bet v1 allergen. Study was performed in Southern Poland, in 2017–2019. Pollen material was collected at 20 sites, of highly or less polluted areas. Protein composition was analysed by SDS-PAGE, while the concentration of Bet v1 was evaluated by ELISA. The obtained results were estimated at the background of the particulate matter (PM10) level and the birch pollen seasons in Kraków. The electrophoregrams of pollen samples collected at different sites showed huge differences in staining intensities of individual protein subunits, also among important birch allergens: Bet v1, Bet v2, Bet v6 and Bet v7. The level of Bet v1 was significantly higher in the pollen samples collected at the more polluted sites. While the birch pollen allergenic potential is determined, the both pollen exposure and the content of the main allergenic components should be considered, as factors causing immunological response and clinical symptoms manifestation in sensitive individuals.