Pollen grains of queen sago (Cycas circinalis L.), a source of aeroallergen from West Bengal, India: an immunochemical approach

Cycas circinalis L. or queen sago is a common ornamental gymnosperm in tropics and subtropics. The objectives of the study were (a) to observe the seasonal variation of queen sago pollen in the atmosphere of a rural and an industrial area of West Bengal, India, (b) to visualize its allergenic potential on local population, and (c) to identify and isolate the important IgE-binding protein component present in the pollen extract. A two-year aerobiological survey was performed with Burkard personal volumetric sampler, and Cycas pollen was found to be present in air during April–July. Among 172 respiratory allergic patients of study area, 25.58% showed skin reaction to Cycas pollen extract. The allergenicity of the pollen extract was confirmed by in vivo (skin reaction test) and in vitro (IgE-ELISA and dot blotting) analyses and immunoblotting. Two components of 39.6 and 20.7 kDa were found to be the important IgE-binding proteins in pollen extract. The 20.7 kDa component was purified by two-step gel electrophoresis and it was found to retain its IgE reactivity. This component can be used for further work in diagnostic and therapeutic purpose in susceptible individuals. The overall study demonstrated that the pollen grains of Cycas circinalis is one of the important aeroallergen source of West Bengal, India,     

<Emphasis Type="Italic">Chenopodium album</Emphasis> pollen profilin (Che a 2): homology modeling and evaluation of cross-reactivity with allergenic profilins based on predicted potential IgE epitopes and IgE reactivity analysis

The inhalation of Chenopodium album (C. album) pollen has been reported as an important cause of allergic respiratory symptoms. The aim of this study was to produce the recombinant profilin of C. album (rChe a 2) pollen and to investigate its cross-reactivity with other plant-derived profilins based on potential conformational epitopes and IgE reactivity analysis. Che a 2-coding sequence was cloned, expressed, and purified using one step metal affinity chromatography to recover high-purity target protein. We assessed cross-reactivity and predicted IgE potential epitopes among rChe a 2 and other plant-derived profilins. Immunodetection and inhibition assays using sixteen individual sera from C. album allergic patients demonstrated that purified rChe a 2 could be the same as that in the crude extract. The results of inhibition assays among rChe a 2 and other plant-derived profilins were in accordance with those of the homology of predicted conserved conformational regions. In this study, amino acid sequence homology analysis showed that a high degree of IgE cross-reactivity among plant-derived profilins may depend on predicted potential IgE epitopes.     

Airborne Poaceae pollen in Porto (Portugal) and allergenic profiles of several grass pollen types

This work aims to investigate the presence of airborne grass pollen and to identify antigenic and allergenic profiles from eight different grass species collected in the Porto region (Portugal). Poaceae airborne pollen, sampled using a Hirst-type volumetric trap during 2003–2007, was the second most abundant type, and high concentrations were found from April to August. Pollen proteins extracted from the eight grass species collected were separated by SDS-PAGE, being the allergenic profile investigated by immunoblotting using sera from atopic patients and maize profilin polyclonal antibody (ZmPRO3). Pollen extract profiles showed several bands ranging from 10 to 97 kDa. In immunoblotting studies, a low molecular weight protein (12–13 kDa) was recognized by profilin antibody. Also, in all pollen extracts except Zea mays, the IgE binding proteins of 12–13 kDa were detected in sera from the 25 patients with different sensitization profiles presenting high IgE values (>80 kU/l). This protein can be considered as a potential causal agent of the allergic respiratory diseases.     

Assessment of airborne Asteraceae pollen in Brisbane, Australia

In the southern hemisphere, the airborne prevalence and seasonal distribution of Asteraceae pollen are poorly characterized. In this study, we assessed the longitudinal distribution of Asteraceae pollen using an historic aerobiological dataset. The influence of weather parameters on pollen derived from Asteraceae species was additionally explored. Asteraceae pollen was collected using a static Burkard 7-day spore trap during a 5-year (June 1994–May 1999) aerometric surveillance program in Brisbane, Australia. Two discrete seasonal intervals were identified each sampling year. The first interval (September and October) consisted of small quantities of unidentifiable Asteraceae pollen. The second interval (March and April) was characterized by the highest concentrations of Asteraceae pollen that were predominantly derived from Baccharis halimifolia. Associations between daily Asteraceae pollen concentrations and temperature were observed during each sampling year. The results of the study demonstrated that pollen derived from B. halimifolia, an invasive weed species introduced in the late nineteenth century, continue to be in the environment of Brisbane. Given the public health burden associated with personal Asteraceae pollen exposure in other geographic locations, future Australian-based clinical studies should characterize the contribution of Asteraceae pollen, in particular B. halimifolia, to allergic sensitization and respiratory morbidity.     

Biochemical and immunochemical characterization of hop-hornbeam (Ostrya Carpinifolia Scop.) pollen

In the last few years Ostrya carpinifolia pollen is consideredas an important cause of respiratoryallergy in Mediterranean areas. The concentration ofthe pollen was measured over a period of fifteen yearsfrom 1981 to 1996 in an area around Genoa; the resultsof this study have clearly indicated an increasingtrend that correlate with persons sensitization.In this study we sought to define the immunochemical andbiochemical properties of hop-hornbean pollen. Soluble proteins extracted from Ostryacarpinifolia pollen and from the taxonomicallyrelated species Corylus Avellana, were analyzedby polyacrylamide gel electrophoresis (SDS-PAGE), byhorizontal isoelectrofocusing (IEF) and by twodimensions electrophoresis (2D-PAGE). Allergenicproteins were identified with sera of sensibilizedpatients and cross-reactivity was evaluated byimmunoblotting techniques. The electrophoreticanalysis showed a partial identity between theproteins from Ostrya and Corylus extracts. The immunoblotting assay, developed withhuman IgE from subjects allergic to hop-hornbeampollen, displayed the major IgE reactivity for acomponent with a molecular weight of 17 kDa expressedin both Ostrya and Corylus extracts. This reactivity is consistent with the presence ofBet v 1 that is described as the major pollen allergenin the Betulaceae and Corylaceae families. Sera fromsubjects allergic to Ostrya were then preadsorbed with recombinant Bet v 1 immobilized in the Pharmacia CAP System; a significant reduction ofthe IgE binding activity was observed after thetreatment. We therefore suggest that Bet v 1 couldbe one of the allergenic proteins present in theOstrya pollen possibly being responsible forcross-reactivity with other members of taxonomicallyrelated families.     

Aerobiology, allergenicity and biochemistry of three pollen types in Berhampore town of West Bengal, India

An aerobiological survey was performed in Berhampore town of West Bengal, India, to know the frequency of three common airborne pollen, namely Acacia auriculiformis, Eucalyptus citriodora and Madhuca indica using an ASTIR one day volumetric sampler. Acacia pollen showed its peak concentration in September, followed by Madhuca in April, while Eucalyptus showed its two peaks between September–October and January–April. Meteorological factors like temperature, RH, rainfall played an important role in release and dispersal of pollen. Skin prick tests with the antigenic extracts of the three pollen types, showed their allergenic potentialities. The highest markedly positive reactions were exhibited by Eucalyptus (34.04%), followed by Madhuca (22.93%) and Acacia (21.87%). 30–60% (NH₄)₂ SO₄ cut fraction (Fraction II) of each pollen type showed maximum positivity in skin prick test. Biochemical analysis showed that Acacia pollen was richer in protein and carbohydrate, than the other two types. The total protein component of the above types were studied by SDS-PAGE showing different protein bands with a range of molecular weight 29–110 KD. In isolated fraction II (allergenically most potent) of Eucalyptus and Madhuca different protein band of 43–200 KD were obtained, while a single protein band of 57 KD was obtained for Acacia. The IgE specific allergenic reactivity was confirmed by Dot-blotting technique. This revised version was published online in June 2006 with corrections to the Cover Date.     

Minor pollinosis fromCryptomeria japonica in Varese,northern Italy. Aerobiological monitoring and main clinical features

In the last 2 years, we have conducted an aerobiological monitoring program ofCryptomeria japonica, a plant belonging to the family of Taxodiaceae that sometimes causes pollinosis in the period from February to April. Throughout 1994, we checked the incidence of its sensitization and the clinical effects in 85 subjects with correlated seasonal symptoms, who gave a positive skin prick test (SPT) for Betulaceae and/or Corylaceae. Twenty-five patients (29.4%; 19 M; 6 F; mean age, 38.8 years) all with oculorhinitis, were SPT positive to an allergenic extract ofCryptomeria. RAST confirmed this positivity in 44% of the cases. No patients showed monosensitization forCryptomeria to either SPT or RAST. Two subjects gave a positive result on specific nasal provocation. RAST inhibition showed no cross-reaction betweenCryptomeria and birch pollen. During the pollen season each patient made a list, scoring symptoms and specifying any drugs used, so we could correlate these elements with aerobiological observations. The pollen concentration probably exceeded the allergizing threshold forCryptomeria on 8 days during 3 months of recording. The intensity and duration of symptoms seemed to be mainly influenced by sensitization to Betulaceae and Corylaceae. It is thus possible that a combination of ‘minor’ pollinosis may produce seasonal symptoms in allergic patients.     

nCup a 1 as a marker of allergy to cypress pollen

The increase in polysensitisations among allergic patients has led us to search for suitable means of diagnosis for identifying true sensitisation, and distinguishing true sensitisation from cross-reactivity. Cross-reactive carbohydrate determinants (CCDs) present in glycoproteins from cypress pollen extracts have been linked with such cross-reactivity, particularly in in vitro assays. The application of component-resolved diagnosis using recombinant allergens makes it possible to identify true allergens. The problem arises when the allergen available for the usual diagnostic methods, which are used as a reference for the diagnosis of allergy to cypress pollen nCup a 1, is a native allergen. The aim of the study was to validate the native allergen nCup a as a marker of true sensitisation to cypress pollen. The sera of 96 subjects with a proven allergy to cypress pollen were analysed. We then quantified IgE specific to Cupressus arizonica and to nCup a 1 and also analysed the CCDs in subjects sensitised to several tree pollen allergens, presenting with MUXF3-specific IgE. Results revealed that there is a statistically significant correlation between conventional diagnostic techniques used to determine allergy to cypress pollen (SPT and IgE Cupressus arizonica) and sensitisation to nCup a 1. CCD quantification in subjects sensitised to several tree pollen antigens showed that these did not interfere with our results. We validated the native Cupressus arizonica allergen, nCup a 1, as a marker of allergy to cypress pollen in our population.     

Allergenicity of Acroptilon repens and Juglans regia pollen in rats

Pollen proteins that are located in the cytoplasm or on the surface of the exine can function as allergens and evoke immune system responses in sensitive patients, leading to allergic rhinitis and asthma. In this research, the pollen allergenicity and ability to induce IgE response of the pollen of two plant species were studied in rats. Acroptilon repens is an herbaceous, invasive plant with entomophilous pollen, while Juglans regia which is a tree crop produces anemophilous pollen. Immunoblot analysis using sera of sensitised rats revealed IgE reactivity to three protein bands including the 70, 41 and 25.12 kDa bands present in the A. repens pollen extract, while only one single immunogenic band of 11 kDa was detected in J. regia pollen extract. Both pollen extracts increased the eosinophil content and caused some clinical signs of allergy in treated rats. The results showed that both entomophilous and anemophilous pollen can be allergenic.     

Allergy symptoms in relation to alder and birch pollen concentrations in Finland

In order to study allergic people responding to daily changes in pollen concentrations, we compared personal diary data on allergic symptoms and the use of allergy medicines to daily pollen counts during the two unequal alder and birch pollen seasons of 2009 and 2010. Almost 90% of the 61 subjects with physician-diagnosed birch pollinosis developed conjunctival, nasal or other symptoms during the peak birch pollination. Most subjects (95%) also reported symptoms during the alder pollination. Despite a delay between the most severe symptoms and the pollen peaks and the increased risk of allergy symptoms between the alder and birch pollen peaks at much lower pollen concentrations, the number of subjects with allergy symptoms correlated with the daily pollen concentrations in both years (r ₀₉ = 0.35, r ₁₀ = 0.36, p < 0.01). The positive correlation was even stronger (r ₀₉ = 0.69, r ₁₀ = 0.74, p < 0.001) in relation to the cumulative sum of daily concentrations. The use of allergy medicines precisely followed the abundance of allergy symptoms in both years (r ₀₉ = 0.96, r ₁₀ = 0.70, p < 0.001). We conclude that there is a fair correlation between the daily allergy symptoms and the particular pollen concentrations, but the risk of developing symptoms at low, moderate and high concentrations is affected by the progression of the pollen season.     

Allergenic pollen and pollinosis in Warsaw

Summary Our study of fifty two hay fever patients included twenty six solely allergic to grass pollen and twenty six exhibiting allergy to various pollen species, such as hazel, birch, oak, poplar, andArtemisia. Their total and specific IgE response was evalutated by the immunoenzymatic method, while clinical reactivity was assessed by recording nasal and bronchial symptom scores between mid-March and mid-July. Simultaneously pollen counts were made. Polysensitized patients showed significantly higher levels of both total and specific IgE, which testifies to the enhanced quantitative and qualitative IgE. Multisensitized patients reacted earlier than patients sensitized to grass pollen only, which confirms that non-grass plants flowering only in the spring cause the priming effect on the nasal and bronchial mucosa. The early symptoms may be attributable to tree pollen sensitivity or may refletct higher grass pollen IgE levels in the polysensitized group. Characteristically, nasal symptoms preceded bronchial symptoms of several weeks.On comparing nasal washing from the polysensitized patients to washing from patients with grass pollen, we found much cytological material with the predominance of eosinophils.     

Pollen and Mould Allergy in Southern Sardinia (Italy): Comparison of Skin-Test Frequencies and Air Sampling Data

A study was carried out to investigate the influence of atmospheric pollen and fungi in determining allergic diseases by comparing the frequency of skin reactions to air sampling data over a 6-year period.

48% of our population reacted to at least one of the pollen and fungal extracts used. Among pollen, Gramineae gave the most frequent positive reactions, followed by Parietaria, Olea and Compositae. The most common positive skin tests in fungus sensitive patients were by extracts of Alternaria, Cladosporium, Aspergillus and Candida

As for the aerobiological survey, the general trend of pollen and molds was similar during the sampling period. The annual pollen catch did not show remarkable differences during the years sampled, whereas the total fungal spore count was highest in 1988 and 1990.

A comparison between aerobiological and clinical data revealed a good degree of concordance between total pollen counts and positive skin test frequencies for Urticaceae, Gramineae and Oleaceae but not for Compositae (high positive skin reactions and very low counts) and Cupressaceae (high counts and few skin reactions).

A less marked correlation has been found between fungal spore counts and positive skin-test frequencies as compared to pollen. Spores such as Cladosporium, which are present in large number in the air, appear to be less sensitizing, while certain spore types (e.g. Alternaria), seem to be able to sensitize patients in spite of their low atmospheric concentrations.     

Allergen Microarray Indicates Pooideae Sensitization in Brazilian Grass Pollen Allergic Patients

Background

Grass pollen, in particular from Lolium multiflorum is a major allergen source in temperate climate zones of Southern Brazil. The IgE sensitization profile of Brazilian grass pollen allergic patients to individual allergen molecules has not been analyzed yet.

Objective

To analyze the IgE sensitization profile of a Brazilian grass pollen allergic population using individual allergen molecules.

Methods

We analyzed sera from 78 grass pollen allergic patients for the presence of IgE antibodies specific for 103 purified micro-arrayed natural and recombinant allergens by chip technology. IgE-ELISA inhibition experiments with Lolium multiflorum, Phleum pratense extracts and a recombinant fusion protein consisting of Phl p 1, Phl p 2, Phl p 5 and Phl p 6 were performed to investigate cross-reactivities.

Results

Within the Brazilian grass pollen allergic patients, the most frequently recognized allergens were Phl p 1 (95%), Phl p 5 (82%), Phl p 2 (76%) followed by Phl p 4 (64%), Phl p 6 (45%), Phl p 11 (18%) and Phl p 12 (18%). Most patients were sensitized only to grass pollen allergens but not to allergens from other sources. A high degree of IgE cross-reactivity between Phleum pratense, Lolium multiflorum and the recombinant timothy grass fusion protein was found.

Conclusions

Component-resolved analysis of sera from Brazilian grass pollen allergic patients reveals an IgE recognition profile compatible with a typical Pooideae sensitization. The high degree of cross-reactivity between Phleum pratense and Lolium multiflorum allergens suggests that diagnosis and immunotherapy can be achieved with timothy grass pollen allergens in the studied population.     

Minor pollinosis fromCryptomeria japonica in Varese,northern Italy. Aerobiological monitoring and main clinical features

In the last 2 years we have conducted an aerobiological monitoring program ofCryptomeria japonica, a plant belonging to the family of Taxodiaceae that sometimes causes pollinosis in the period from February to April. Throughout 1994, we checked the incidence of its sensitization and the clinical effects in 85 subjects with correlated seasonal symptoms, who gave a positive skin prick test (SPT) for Betulaceae and/or Corylaceae. Twenty-five patients (29.4%; 19 M; 6 F; mean age, 38.8 years) all with oculorhinitis, were SPT positive to an allergenic extract ofCryptomeria. RAST confirmed this positivity in 44% of the cases. No patients showed monosensitization forCryptomeria to either SPT or RAST. Two subjects gave a positive result on specific nasal provocation. RAST inhibition showed no cross-reaction betweenCryptomeria and birch pollen. During the pollen season each patient made a list, scoring symptoms and specifying any drugs used, so we could correlate these elements with aerobiological observations. The pollen concentration probably exceeded the allergizing threshold forCryptomeria on 8 days during 3 months of recording. The intensity and duration of symptoms seemed to be mainly influenced by sensitization to Betulaceae and Corylaceae. It is thus possible that a combination of minor pollinosis may produce seasonal symptoms in allergic patients.     

Quantitative dynamics of Ambrosia pollen grains in Bulgaria

The aerobiological study of the Ambrosia pollen in Sofia, Bulgaria began in 1991. The quantitative dynamics of Ambrosia pollen grains during the last seven years was determined by Chebishev's method of parabolic interpolation with orthogonal polynoms. Thesensitization to ragweed pollen was investigated in random groups of patients with clinical manifestation of hay fever.This investigation proves the previouslyobserved tendency of an increase in thespreading of Ambrosia pollen in Bulgaria.     

Prevalence of allergenic pollen grains in the aerosol of the city of Calcutta,India: a two year study

The role of pollen grains as a causative agent of respiratory allergic disorders such as asthma and allergic rhinitis is common and very well established. The aim of this study was to assess the frequencies of airborne pollen in the Calcutta metropolis and to identify the taxa which cause significant amounts of sensitization. An aeropalynological survey of the atmosphere of Calcutta was carried out from 2004 to 2006. Skin tests were performed with a panel of the most common pollen types on local patients with clinical features of pollinosis. The meteorological factors responsible for the frequency of the pollen types were analysed. The results of monthly visits to the clinic by these patients were correlated with the monthly pollen counts of three dominant and perennial pollen taxa. The dominant pollen types were Trema (19%), Poaceae (12.98%), Casuarina (5.76%), Cocos (5.7%), Azadirachta (4.65%), Peltophorum (3.71%), Cyperaceae (3.68%), Delonix (3.18%) and Areca (2.56%). Total pollen concentration seems to have a significant positive correlation with temperature and wind speed whereas there was a negative correlation with humidity. Skin tests were most frequently found to be positive with the pollen of Poaceae (49%), Azadirachta (46%), Cocos (47%), Cyperaceae (35%), Peltophorum (33%), Areca (29%), Phoenix (26%), and Borassus (23%). A positive correlation occurred between visits to the clinic and monthly pollen count of Areca, Cocos, and Poaceae. This is the first study to design a pollen calendar for Calcutta city; it will provide useful data for enabling allergologists to achieve accurate diagnosis for patients with pollen hypersensitivity.     

Airborne pollen patterns in Mar del Plata atmosphere (Argentina) and its relationship with meteorological conditions

This work is part of a series of aerobiological researches conducted in the city of Mar del Plata, Argentina. The annual, seasonal and daily features of the pollen cloud were analyzed over 2 years, together with the effects of the meteorological variables. Cupressaceae amounted to 75 and 54% of the annual total in each year, and it was analyzed separately due to its proven overrepresentation. Eighty-five pollen types were registered during the 2 years, bearing a similar annual pattern (more than 50% from August to November, December also being an important month). Arboreal pollen predominated between late winter and spring (61 and 49% of the annual total), while non-arboreal pollen did so from late spring to summer. The observed seasonality was significantly correlated with the monthly mean temperatures and associated with its effect on phenology. No significant differences along the day were detected between years and 50% of the daily total was recorded between 1000 and 1600 hours. The maximum concentration was registered at 1000 hours in spring and summer, but later in autumn and winter. Regarding the hourly scale, the significant correlation of total pollen with temperature and wind speed was positive, albeit negative with relative humidity. These effects are linked to airborne pollen release and transport. The role rainfall plays on airborne pollen is discussed. To interpret the results, it is important to consider the time scale at which the aerobiological phenomenon is analyzed.     

Scanning and transmission electron microscopy related to immunochemical analysis of grass pollen

The exine stereostructure (scanning electron microscopy) and ultrastructure (transmission electron microscopy) of pollen of seven grass species, is related to the allergens extracted from these pollen grains. The heterogeneity of the allergens was studied by the immunoprint technique and revealed by labelling the binding of grass pollen sensitive patients IgE antibodies. Using patient sera recognizing a very restricted number of allergens, we showed that a group of pollen had a great number of allergens in common (Dactylis, Agrostis, Festuca, Lolium, Holcus) and, in decreasing cross reactivities order, we found Avena and, finally, Zea mays. The tectum stereostructure shows presence of insulae in all pollen grains except in Zea mays which has small isolated spinules. These insulae are separated by very wide and deep interinsular spaces in Avena sativa with connections between insulae. In the remaining species, no connections were seen between the insulae. These observations were in good correlation with the immunological cross-reactivity of the allergens present in the pollen. In all species, there are microperforations in the bottom of the interinsular spaces, which are the opening of the tectal microchannels.