IgE cross-reactivity between Pas n 1 of Bahia grass pollen and other group 1 grass pollen allergens

Background The subtropical Bahia grass (Paspalum notatum) is an important source of pollen allergens with an extended season of pollination and wide distribution in warmer climates. The immunological relationship between pollen allergens of Bahia grass and temperate grasses is unresolved. Methods Serum IgE reactivity of grass pollen-allergic patients with Bahia, Ryegrass and Bermuda grass pollen extracts and their purified group 1 allergens, Pas n 1, Lol p 1 and Cyn d 1, were compared by immunoblotting, ELISA, inhibition ELISA, basophil activation by flow cytometry and molecular modeling. Results Differences in antibody recognition of allergenic components between Bahia grass and Ryegrass pollen were observed by immunoblotting. Eight grass pollen-allergic patients from a temperate region showed greater serum IgE reactivity with Ryegrass pollen than Bahia grass by ELISA. For seven of these sera, Ryegrass pollen inhibited IgE reactivity with Bahia grass pollen but not the converse. For 51 sera from grass pollen-allergic patients in this temperate region, IgE reactivity with Lol p 1 was greater than Pas n 1 or Cyn d 1. IgE reactivity with Lol p 1 was not inhibited by Pas n 1 or Cyn d 1, but Pas n 1 IgE reactivity was inhibited by Lol p 1. Two group 1 grass pollen allergen-specific mAb distinguished between temperate and subtropical grass pollens. Basophil activation for three patients tested was greater by Ryegrass pollen than Bahia or Bermuda grass, and by Lol p 1 than Pas n 1 or Cyn d 1. In contrast, two patients from a subtropical region had higher serum IgE reactivity with Bahia grass pollen than Ryegrass and Bahia grass pollen inhibited IgE reactivity with Ryegrass. A structural model of Pas n 1 showed amino acids implicated in IgE epitopes of other group 1 allergens were juxtaposed on the surface. Conclusion Allergens from subtropical Bahia grass pollen, including Pas n 1, share antigenic determinants with temperate grass pollen allergens, but patients exhibit higher serum IgE reactivity to their locally predominant grass pollen. Basophil activation by Bahia grass pollen and Pas n 1 in patients from a temperate climate indicates clinically relevant cross-sensitization between temperate and subtropical grass pollens.

A 30-Day-Ahead Forecast Model for Grass Pollen in North London, United Kingdom

A 30-day ahead forecast method has been developed for grass pollen at north London. The total period of the grass pollen season is covered by eight multiple regression models, each covering a 10-day period running consecutively from 21st May to 8th August. This means that three models were used for each 30-day forecast. The forecast models were produced using grass pollen and environmental data from 1961-1999 and tested on data from 2000 and 2002. Model accuracy was judged in two ways: the number of times the forecast model was able to successfully predict the severity (relative to the 1961-1999 dataset as a whole) of grass pollen counts in each of the eight forecast periods on a scale of one to four; and the number of times the forecast model was able to predict whether grass pollen counts were higher or lower than the mean. The models achieved 62.5% accuracy in both assessment years when predicting the relative severity of grass pollen counts on a scale of one to four, which equates to six of the eight 10-day periods being forecast correctly. The models attained 87.5% and 100% accuracy in 2000 and 2002 respectively when predicting whether grass pollen counts would be higher or lower than the mean. Attempting to predict pollen counts during distinct 10-day periods throughout the grass pollen season is a novel approach. The models also employed original methodology in the use of winter averages of the North Atlantic Oscillation to forecast 10-day means of allergenic pollen counts.

Constructing a 7-day Ahead Forecast Model for Grass Pollen at North London, United Kingdom

A number of media outlets now issue medium-range (~7 day) weather forecasts on a regular basis. It is therefore logical that aerobiologists should attempt to produce medium-range forecasts for allergenic pollen that cover the same time period as the weather forecasts. The objective of this study is to construct a medium-range (< 7 day) forecast model for grass pollen at north London. The forecast models were produced using regression analysis based on grass pollen and meteorological data from 1990-1999 and tested on data from 2000 and 2002. The modelling process was improved by dividing the grass pollen season into three periods; the pre-peak, peak and post peak periods of grass pollen release. The forecast consisted of five regression models. Two simple linear regression models predicting the start and end date of the peak period, and three multiple regression models forecasting daily average grass pollen counts in the pre-peak, peak and post-peak periods. Overall the forecast models achieved 62% accuracy in 2000 and 47% in 2002, reflecting the fact that the 2002 grass pollen season was of a higher magnitude than any of the other seasons included in the analysis. This study has the potential to make a notable contribution to the field of aerobiology. Winter averages of the North Atlantic Oscillation were used to predict certain characteristics of the grass pollen season, which presents an important advance in aerobiological work. The ability to predict allergenic pollen counts for a period between five and seven days will benefit allergy sufferers. Furthermore, medium-range forecasts for allergenic pollen will be of assistance to the medical profession, including allergists planning treatment and physicians scheduling clinical trials.

Examining High Magnitude Grass Pollen Episodes at Worcester, United Kingdom, Using Back-Trajectory Analysis

Trajectory analysis is a valuable tool that has been used before in aerobiological studies, to investigate the movement of airborne pollen. This study has employed back-trajectories to examine the four highest grass pollen episodes at Worcester, during the 2001 grass pollen season. The results have shown that the highest grass pollen counts of the 2001 season were reached when air masses arrived from a westerly direction. Back-trajectory analysis has a limited value to forecasters because the method is retrospective and cannot be employed directly for forecasting. However, when used in conjunction with meteorological data this technique can be used to examine high magnitude events in order to identify conditions that lead to high pollen counts.

Characteristics of Grass Pollen Seasons in Areas of Southern Spain and the United Kingdom

Spatial and temporal variations in daily grass pollen counts and weather variables are described for two regions with different bio-geographical and climatic regimes, southern Spain and the United Kingdom. Daily average grass pollen counts are considered from six pollen-monitoring sites, three in southern Spain (Ciudad Real, Córdoba and Priego) and three in the United Kingdom (Edinburgh, Worcester and Cambridge). Analysis shows that rainfall and maximum temperatures are important factors controlling the magnitude of the grass pollen season in both southern Spain and the United Kingdom, and that the strength and direction of the influence exerted by these variables varies with geographical location and time.

Temporal and Georaphical Variations in Grass Pollen Seasons in Areas of Western Europe: an Analysis of Season dates at Sites of the European Pollen Information System.

Geographical and temporal variations in the start dates of grass pollen seasons are described for selected sites of the European Pollen Information Service. Daily average grass pollen counts are derived from Network sites in Finland, the Netherlands, Denmark, United Kingdom, Austria, Italy and Spain, giving a broad longitudinal transect over Western Europe. The study is part of a larger project that also examines annual and regional variations in the severity, timing of the peak and duration of the grass pollen seasons. For several sites, data are available for over twenty years enabling long term trends to be discerned. The analyses show notable contrasts in the progression of the seasons annually with differing lag times occurring between southern and northern sites in various years depending on the weather conditions. The patterns identified provide some insight into geographical differences and temporal trends in the incidence of pollinosis. The paper discusses the main difficulties involved in this type of analysis and notes possibilities for using data from the European Pollen Information service to construct pan European predictive models for pollen seasons.

Influence of the North Atlantic Oscillation on Grass Pollen Counts in Europe

Relationships between temporal variations in the North Atlantic Oscillation (NAO) and grass pollen counts at 13 sites in Europe, ranging from Córdoba in the South-West and Turku in the North-East, were studied in order to determine spatial differences in the amount of influence exerted by the NAO on the timing and magnitude of grass pollen seasons. There were a number of significant (p<0.05) relationships between the NAO and start dates of the grass pollen season at the 13 pollen-monitoring sites. The strongest associations were generally recorded near to the Atlantic coast. Several significant correlations also existed between winter averages of the NAO and grass pollen season severity. Traditional methods for predicting the start or magnitude of grass pollen seasons have centred on the use of local meteorological observations, but this study has shown the importance of considering large-scale patterns of climate variability like the NAO.

Long-term and Short-term Forecast Models for Poaceae (Grass) Pollen in Poznan, Poland, Constructed Using Regression Analysis

Airborne concentrations of Poaceae pollen have been monitored in Poznań for more than ten years and the length of the dataset is now considered sufficient for statistical analysis. The objective of this paper is to produce long-range forecasts that predict certain characteristics of the grass pollen season (such as the start, peak and end dates of the grass pollen season) as well as short-term forecasts that predict daily variations in grass pollen counts for the next day or next few days throughout the main grass pollen season. The method of forecasting was regression analysis. Correlation analysis was used to examine the relationship between grass pollen counts and the factors that affect its production, release and dispersal. The models were constructed with data from 1994-2004 and tested on data from 2005 and 2006. The forecast models predicted the start of the grass pollen season to within 2 days and achieved 61% and 70% accuracy on a scale of 1-4 when forecasting variations in daily grass pollen counts in 2005 and 2006 respectively. This study has emphasised how important the weather during the few weeks or months preceding pollination is to grass pollen production, and draws attention to the importance of considering large-scale patterns of climate variability (indices of the North Atlantic Oscillation) when constructing forecast models for allergenic pollen.

Do Urban Canyons Influence Street Level Grass Pollen Concentrations

In epidemiological studies, outdoor exposure to pollen is typically estimated using rooftop monitoring station data, whilst exposure overwhelmingly occurs at street level. In this study the relationship between street level and roof level grass pollen concentrations was investigated for city centre street canyon environments in Aarhus, Denmark, and London, UK, during the grass pollen seasons of 2010 and 2011 respectively. For the period mid-day to late evening, street level concentrations in both cities tended to be lower than roof-level concentrations, though this difference was found to be statistically significant only in London. The ratio of street/roof level concentrations was compared with temperature, relative humidity, wind speed and direction, and solar radiation. Results indicated that the concentration ratio responds to wind direction with respect to relative canyon orientation and local source distribution. In the London study, an increase in relative humidity was linked to a significant decrease in street/roof level concentration ratio, and a possible causative mechanism involving moisture mediated pollen grain buoyancy is proposed. Relationships with the other weather variables were not found to be significant in either location. These results suggest a tendency for monitoring station data to overestimate exposure in the canyon environment.

Personal Exposure to Grass Pollen: Relating Inhaled Dose to Background Concentration

Background Very few studies on human exposure to allergenic pollen have been conducted using direct methods, with background concentrations measured at city center monitoring stations typically taken as a proxy for exposure despite the inhomogeneous nature of atmospheric pollen concentrations. A 2003 World Health Organization report highlighted the need for an improved understanding of the relation between monitoring station data and actual exposure. Objective To investigate the relation between grass pollen dose and background concentrations measured at a monitoring station, to assess the fidelity of monitoring station data as a qualitative proxy for dose, and to evaluate the ratio of dose rate to background concentration. Methods Grass pollen dose data were collected in Aarhus, Denmark, in an area where grass pollen sources were prevalent, using Nasal Air Samplers. Sample collection lasted for approximately 25 to 30 minutes and was performed at 2-hour intervals from noon to midevening under moderate exercise by 2 individuals. Results A median ratio of dose rate to background concentration of 0.018 was recorded, with higher ratio values frequently occurring at 12 to 2 pm, the time of day when grass species likely to be present in the area are expected to flower. From 4 to 8 pm, dose rate and background concentration data were found to be strongly and significantly correlated (rs = 0.81). Averaged dose rate and background concentration data showed opposing temporal trends. Conclusion Where local emissions are not a factor, background concentration data constitute a good quantitative proxy for inhaled dose. The present ratio of dose rate to background concentration may aid the study of dose–response relations.

Seasonal Variation in Diurnal Atmospheric Grass Pollen Concentration Profiles

In this study, the diurnal atmospheric grass pollen concentration profile within the Danish city of Aarhus was shown to change in a systematic manner as the pollen season progressed. Although diurnal grass pollen profiles can differ greatly from day-to-day, it is common practice to establish the time of day when peak concentrations are most likely to occur using seasonally averaged diurnal profiles. Atmospheric pollen loads are highly dependent upon emissions, and different species of grass are known to flower and emit pollen at different times of the day and during different periods of the pollen season. Pollen concentrations are also influenced by meteorological factors – directly through those parameters that govern pollen dispersion and transport, and indirectly through the weather-driven flowering process. We found that three different profiles dominated the grass pollen season in Aarhus – a twin peak profile during the early season, a single evening profile during the middle of the season, and a single midday peak during the late season. Whilst this variation could not be explained by meteorological factors, no inconsistencies were found with the theory that it was driven by a succession of different grass species with different diurnal flowering patterns dominating atmospheric pollen loads as the season progressed. The potential for exposure was found to be significantly greater during the late-season period than during either the early- or mid-season periods.

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.

Anti-idiotypic Fab Fragments Image a Conserved N-terminal Epitope Patch of Grass Pollen Allergen Phl p 1

BACKGROUND AND AIMS: Naturally occurring anti-idiotypic antibodies structurally mimic the original antibody epitope. Anti-idiotypes, therefore, are interesting tools for the portrayal of conformational B-cell epitopes of allergens. In this study we used this strategy particularly for major timothy grass pollen (Phleum pratense) allergen Phl p 1. METHODS AND RESULTS: We used a combinatorial phage display library constructed from the peripheral IgG repertoire of a grass pollen allergic patient which was supposed to contain anti-idiotypic Fab specificities. Using purified anti-Phl p 1 IgG for biopanning, several Fab displaying phage clones could be isolated. 100 amplified colonies were screened for their binding capacity to anti-Phl p 1-specific antibodies, finally resulting in four distinct Fab clones according to sequence analysis. Interestingly, heavy chains of all clones derived from the same germ line sequence and showed high homology in their CDRs. Projecting their sequence information on the surface of the natural allergen Phl p 1 (PDB ID: 1N10) indicated matches on the N-terminal domain of the homo-dimeric allergen, including the bridging region between the two monomers. The resulting epitope patches were formed by spatially distant sections of the primary allergen sequence. CONCLUSION: In this study we report that anti-idiotypic specificities towards anti-Phl p 1 IgG, selected from a Fab library of a grass pollen allergic patient, mimic a conformational epitope patch being distinct from a previously reported IgE epitope area.

Molecular cloning, expression and immunological characterisation of Pas n 1, the major allergen of Bahia grass Paspalum notatum pollen

Bahia grass, Paspalum notatum, is a clinically important subtropical grass with a prolonged pollination season from spring to autumn. We aimed to clone and characterise the major Bahia grass pollen allergen, Pas n 1. Grass pollen-allergic patients presenting to a tertiary hospital allergy clinic were tested for IgE reactivity with Bahia grass pollen extract by skin prick testing, ImmunoCAP, ELISA and immunoblotting. Using primers deduced from the N-terminal peptide sequence of a group 1 allergen of Bahia grass pollen extract separated by two-dimensional gel electrophoresis, the complete Pas n 1 cDNA was obtained by rapid amplification of cDNA ends and cloned. Biological relevance of recombinant Pas n 1 expressed in Escherichia coli was assessed by serum IgE reactivity and basophil activation. Twenty-nine of 34 (85%) consecutive patients presenting with grass pollen allergy were skin prick test positive to Bahia grass pollen. The Pas n 1 cDNA has sequence homology with the β-expansin 1 glycoprotein family and is more closely related to the maize pollen group 1 allergen (85% identity) than to ryegrass Lol p 1 or Timothy grass Phl p 1 (64 and 66% identity, respectively). rPas n 1 reacted with serum IgE in 47 of 55 (85%) Bahia grass pollen-allergic patients, activated basophils and inhibited serum IgE reactivity with the 29 kDa band of Bahia grass pollen extract. In conclusion the cDNA for the major group 1 allergen of the subtropical Bahia grass pollen, Pas n 1, was identified and cloned. rPas n 1 is immunologically active and is a valuable reagent for diagnosis and specific immunotherapy of grass pollen allergy.

Specific IgE recognition of pollen allergens from subtropic grasses in patients from the subtropics

Background Pollens of subtropical grasses, Bahia (Paspalum notatum), Johnson (Sorghum halepense), and Bermuda (Cynodon dactylon), are common causes of respiratory allergies in subtropical regions worldwide. Objective To evaluate IgE cross-reactivity of grass pollen (GP) found in subtropical and temperate areas. Methods Case and control serum samples from 83 individuals from the subtropical region of Queensland were tested for IgE reactivity with GP extracts by enzyme-linked immunosorbent assay. A randomly sampled subset of 21 serum samples from patients with subtropical GP allergy were examined by ImmunoCAP and cross-inhibition assays. Results Fifty-four patients with allergic rhinitis and GP allergy had higher IgE reactivity with P notatum and C dactylon than with a mixture of 5 temperate GPs. For 90% of 21 GP allergic serum samples, P notatum, S halepense, or C dactylon specific IgE concentrations were higher than temperate GP specific IgE, and GP specific IgE had higher correlations of subtropical GP (r = 0.771-0.950) than temperate GP (r = 0.317-0.677). In most patients (71%-100%), IgE with P notatum, S halepense, or C dactylon GPs was inhibited better by subtropical GP than temperate GP. When the temperate GP mixture achieved 50% inhibition of IgE with subtropical GP, there was a 39- to 67-fold difference in concentrations giving 50% inhibition and significant differences in maximum inhibition for S halepense and P notatum GP relative to temperate GP. Conclusion Patients living in a subtropical region had species specific IgE recognition of subtropical GP. Most GP allergic patients in Queensland would benefit from allergen specific immunotherapy with a standardized content of subtropical GP allergens.