城市中的花粉致敏植物及其影响因素

大量致敏花粉飘散于空气中会引发花粉症,严重危害人体健康。由于不当的城市绿地建设使得花粉致敏植物在城市中大量聚集,加之城市热岛效应和交通污染等多方面的因素,使得城市中花粉症患者数量激增。综述了花粉致敏植物的种类、物候特征和影响因素提出了控制花粉致敏植物的建议;总结了目前花粉致敏植物研究中应进一步关注的问题,旨在为改善城市空气质量,建设合理绿化环境提供科学依据。     

The Effects of Air Pollution on Structures, Proteins and Allergenicity of Pollen Grains

The prevalence of allergic disease has increased world wide during the last decades. Pollen allergy is the most typical form of allergic disease. The increase in its frequency during recent years is the most evident. Environmental factors play an important role in the problem of pollen allergy in large cities. The aim of this research is determination of allergenicity of Canna pollen in polluted and non-polluted conditions, detection of their allergenic proteins and also elucidation of some microscopic effects of air pollutants on pollen structure and proteins. Mature and immature pollen grains of Canna indica were collected from polluted and non-polluted areas. Pollen grains were studied by scanning electron microscopy. Mice were sensitized by injection of pollen extract and an adjuvant for five times. Allergy potency of different pollen extracts were compared by means of skin test, blood eosinophills number and IgE levels in sensitized and treated animals. Pollen proteins were studied by SDS-PGE and allergenic proteins were detected by immunoblotting techniques. Scanning electron microscope study of the pollen grains showed that in polluted areas, air born particles accumulated on the surface of pollen and changed both pollen's shape and pollen's tectum. Also many vesicles were released out of polluted pollen and the pollen material agglomerated on the surface of pollen. SDS-PAGE showed that different proteins exist in mature and immature pollen. In pollen collected from polluted area, some of protein bands between 22 and 45 kDa were disappeared . Also in all polluted pollen grains, protein content of pollen decreased in response to air pollution causing the release of pollen proteins. According to our experiments and regarding induction of allergic symptoms, the polluted pollen is more effective than non-polluted one, and mature pollen has more allergy potency than immature one.     

Proteomic analysis of the major birch allergen Bet v 1 predicts allergenicity for 15 birch species

Pollen of the European and Asian white birch (Betula pendula and B. platyphylla) causes hay fever in humans. The allergenic potency of other birch species is largely unknown. To identify birch trees with a reduced allergenicity, we assessed the immunochemical characteristics of 15 species and two hybrids, representing four subgenera within the genus Betula, while focusing on the major pollen allergen Bet v 1. Antigenic and allergenic profiles of pollen extracts from these species were evaluated by SDS-PAGE and Western blot using pooled sera of birch-allergic individuals. Tryptic digests of the Bet v 1 bands were analyzed by LC-MS(E) to determine the abundance of various Bet v 1 isoforms. Bet v 1 was the most abundant pollen protein across all birch species. LC-MS(E) confirmed that pollen of all species contained a mixture of multiple Bet v 1 isoforms. Considerable differences in Bet v 1 isoform composition exist between birch species. However, isoforms that are predicted to have a high IgE-reactivity prevailed in pollen of all species. Immunoblotting confirmed that all pollen extracts were similar in immune-reactivity, implying that pollen of all birch species is likely to evoke strong allergic reactions.     

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.     

Identification of specific IgE binding proteins in Castor bean (Ricinus communis) pollen obtained from different source materials

Allergenic components of Ricinus communis pollen obtained from different stages of inflorescence, different time intervals, different years and places were studied by immunoblot analysis. Proteins separated by SDS-PAGE and transferred to NC were identified using pooled sera from 15 skin and RAST positive patients. The IgE binding components in M.W. range of 14 to 70 kD were identified. The protein fractions of 70, 66, 64, 60, 50, 45, 36, 22 and 14 kD are the most prominent allergenic bands. Six samples collected during same pollination season from the same place showed similar allergenic profile. Of the samples collected from different stages of inflorescence, pollen of immature buds showed only three bands as compared to 18 from mature buds and flowers. Variability was seen in the IgE binding components of pollen stored for different years and obtained from different geographic regions of India. The IgE binding pattern of fifteen sera were heterogenous. The number of bands identified by different sera varied from 3 to 18. Two protein components of 66 and 36 kD were recognised by 14 (93.3%) of the 15 sera studied. The result suggests that there exists variations in the specific IgE binding pattern in pollen samples of Castor Bean, obtained from difference source materials.     

Simulated environmental criticalities affect transglutaminase of Malus and Corylus pollens having different allergenic potential

Increases in temperature and air pollution influence pollen allergenicity, which is responsible for the dramatic raise in respiratory allergies. To clarify possible underlying mechanisms, an anemophilous pollen (hazel, Corylus avellana), known to be allergenic, and an entomophilous one (apple, Malus domestica), the allergenicity of which was not known, were analysed. The presence also in apple pollen of known fruit allergens and their immunorecognition by serum of an allergic patient were preliminary ascertained, resulting also apple pollen potentially allergenic. Pollens were subjected to simulated stressful conditions, provided by changes in temperature, humidity, and copper and acid rain pollution. In the two pollens exposed to environmental criticalities, viability and germination were negatively affected and different transglutaminase (TGase) gel bands were differently immunodetected with the polyclonal antibody AtPng1p. The enzyme activity increased under stressful treatments and, along with its products, was found to be released outside the pollen with externalisation of TGase being predominant in C. avellana, whose grain presents a different cell wall composition with respect to that of M. domestica. A recombinant plant TGase (AtPng1p) stimulated the secreted phospholipase A(2) (sPLA(2)) activity, that in vivo is present in human mucosa and is involved in inflammation. Similarly, stressed pollen, hazel pollen being the most efficient, stimulated to very different extent sPLA(2) activity and putrescine conjugation to sPLA(2). We propose that externalised pollen TGase could be one of the mediators of pollen allergenicity, especially under environmental stress induced by climate changes.     

Characterization of protein expression of Platanus pollen following exposure to gaseous pollutants and vehicle exhaust particles

Being major ornamental street trees, species of Platanus are widely planted in the Shanghai urban area. A great deal of allergenic Platanus pollen is released from the trees and suspended in the atmosphere during its flowering season, ultimately causing allergic respiratory diseases. Few papers have focused on the distribution of this type of pollen and its expression of allergenic proteins. In order to investigate any differences in protein expression in Platanus pollen following exposure to gaseous and particulate pollutants, a special apparatus was designed. Exposure condition (such as temperature, humidity, and exposure time) of Platanus pollen and gaseous pollutants can be simulated using of this apparatus. Fresh Platanus orientalis pollen, pollutant gases (NO₂, SO₂, NH₃), and typical urban ambient particles (vehicle exhaust particles, VEPs) were mixed in this device to examine possible changes that might occur in ambient airborne urban pollen following exposure to such pollutants. Our results showed that the fresh P. orientalis pollen became swollen, and new kinds of particles could be found on the surface of the pollen grains after exposure to the pollutants. The results of SDS-PAGE showed that five protein bands with molecular weights of 17–19, 34, 61, 82, and 144 kDa, respectively, were detected and gray scale of these brands increased after the pollen exposure to gaseous pollutants. The two-dimensional gel electrophoresis analysis demonstrated that a Platanus pollen allergenic protein (Pla a1, with a molecular weight of 18 kDa) increased in abundance following exposure to pollutant gases and VEPs, implying that air pollutants may exacerbate the allergenicity of pollen.     

Isolation and partial characterization ofCupressus sempervirens allergens

Summary Allergic rhinitis due to cypress pollen is a well-recognized clinical condition that is evermore frequently diagnosed as a winter allergy in the Mediterranean area. Little is known about the allergen composition of its pollen and the dynamics of its immune response. For these reasons IgE-specific antibody distribution was determined and the allergenic pollen characterized. SDS-PAGE analysis of the whole extract revealed more than ten proteic bands ranging from 10 to 90 kD. The major allergen had a molecular weight of 36 kD and the specific IgE antibody was presene in >95% of the sera tested by immunoblotting technique. Two minor allergens were observed at the 43 and 49 kd protein bands. The results obtained with the whole extract were then compared to those of five commercially available preparations.     

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.     

Mountain cedar pollen induces IgE-independent mast cell degranulation, IL-4 production, and intracellular reactive oxygen species generation

Cedar pollens cause severe allergic disease throughout the world. We have previously characterized allergenic pollen glycoproteins from mountain cedar (Juniperus ashei) that bind to allergen-specific immunoglobulin E (IgE). In the present report, we investigated an alternative pathway of mast cell activation by mountain cedar pollen extract through IgE-independent mechanisms. We show that mountain cedar pollen directly induces mast cell serotonin and IL-4 release and enhances release induced by IgE cross-linking. Concomitant with mediator release, high levels of intracellular reactive oxygen species (ROS) were generated, and both ROS and serotonin release were inhibited by anti-oxidants. These findings suggest that alternative mechanisms exist whereby pollen exposure enhances allergic inflammatory mediator release through mechanisms that involve ROS. These mechanisms have the potential for enhancing the allergenic potency of pollens.     

5种裸子植物花粉外壁成分及结构分析比较

利用傅里叶变换红外光谱仪对银杏、日本冷杉等5种裸子植物的花粉外壁成分进行测定分析,结果表明,这5种裸子植物花粉外壁的红外光谱主要由蛋白质、脂类及多糖类物质的特征吸收峰组成。但不同科属间的花粉外壁主要成分含量存在较大差异,其中银杏科的银杏花粉外壁以蛋白质含量最为丰富;松科的雪松花粉外壁以脂类物质较为丰富;杉科的日本冷杉、杉木和日本柳杉花粉外壁成分以多糖类物质为主,但种间花粉外壁成分仍存在差异。采用扫描电镜观察5种裸子植物花粉,显示银杏、柳杉和杉木花粉粒体积较小,不具气囊,银杏花粉粒外壁表面具较均一条纹状纹饰,日本柳杉和杉木花粉粒外壁具颗粒状突起。雪松和日本冷杉花粉粒具气囊,体积较大,花粉外壁分别是粗糙具小穴状纹理以及表面光滑具微穿孔。     

Reduction of antigenicity of Cry j I, major allergen of Japanese cedar pollen, by the attachment of polysaccharides

An attempt was made to mask the allergenic structure of a major allergen protein, Cry j I (CJI), in Japanese cedar pollen using the Maillard-type polysaccharide conjugation. The SDS-PAGE pattern of the CJI-galactomannan conjugate prepared by the Maillard reaction showed broad bands widely distributed from 50 kDa to more than 100 kDa, suggesting the attachment of galactomannan. The competitive enzyme-linked immunosorbent assay showed that the IgE antibody in the sera of cedar pollen-sensitive patients reacted strongly with CJI, while it did not react with the CJI-galactomannan conjugate. This result suggests that the antigenicity of CJI is greatly reduced by the conjugation with galactomannan.     

空气致敏花粉污染研究进展

对空气致敏花粉污染概念的提出 ,空气致敏花粉污染的特点和影响因素 ,花粉采集方法的改进以及空气致敏花粉污染的研究进展等方面进行了总结 ,指出了研究中存在的问题 ,并对研究前景进行了展望     

Study on allergenicity of Thuja orientalis pollen grains in rat

Cupressaceae pollen allergy is an important cause of pollen allergy throughout the world. Prevalence of allergy to Cupressaceae pollen has increased significantly during the winter over the past 3 decades because of extensive planting of cypress trees for different purposes. Thuja orientalis (Cupressaceae) is a naturally grown plant in Iran and is widely cultivated as a common ornamental plant in this country and other ones. Allergenicity of its pollen has been established, but to this day no allergenic component has been detected. The aim of this research is to study allergenicity and evaluate the immunoglobulin E reactivity to T. orientalis pollen extracts. Pollen grains were directly collected from mature male cones of trees. Pollen proteins were extracted and were analyzed by SDS-PAGE. Total protein content of pollen extracts was measured by Bradford assay. Immunoblotting using the serum of sensitized rats showed a single immunogenic band at about 44KD in pollen extracts. Result of this research proved that pollen grains of T. orientalis are allergenic.     

Variable expression of pathogenesis-related protein allergen in mountain cedar (Juniperus ashei) pollen

Allergic diseases have been increasing in industrialized countries. The environment is thought to have both direct and indirect modulatory effects on disease pathogenesis, including alterating on the allergenicity of pollens. Certain plant proteins known as pathogenesis-related proteins appear to be up-regulated by certain environmental conditions, including pollutants, and some have emerged as important allergens. Thus, the prospect of environmentally regulated expression of plant-derived allergens becomes yet another potential environmental influence on allergic disease. We have identified a novel pathogenesis-related protein allergen, Jun a 3, from mountain cedar (Juniperus ashei) pollen. The serum IgE from patients with hypersensitivity to either mountain cedar or Japanese cedar were shown to bind to native and recombinant Jun a 3 in Western blot analysis and ELISA. Jun a 3 is homologous to members of the thaumatin-like pathogenesis-related (PR-5) plant protein family. The amounts of Jun a 3 extracted from mountain cedar pollen varied up to 5-fold in lots of pollen collected from the same region in different years and between different regions during the same year. Thus, Jun a 3 may contribute not only to the overall allergenicity of mountain cedar pollen, but variable levels of Jun a 3 may alter the allergenic potency of pollens produced under different environmental conditions.     

Hypersensitivity to Ailanthus altissima (tree of heaven) pollen: identification of a major IgE-binding component

Ailanthus altissima or Ailanthus glandulosa (Simaroubaceae) is known as tree of heaven. It is a dioecious plant with staminate and pistillate flowers that grow on separate trees. In recent years, A. altissima has been frequently planted in numerous areas, especially in arid and semiarid lands of Iran and also used as an ornamental tree in several Iran cities including Kerman. The aim of this research was to identify IgE-binding proteins responsible for type I hypersensitivities of A. altissima pollen extract. In this study, pollen’s proteins were extracted and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Total protein content of pollen extracts was measured by Bradford assay. Allergenicity of pollen extract was evaluated by skin test and immunoblotting. Results showed that total protein concentration of A. altissima pollen was found to be 1.34 μg μl^?1. In A. altissima pollen extract, different protein fractions were identified by SDS-PAGE mostly at 42, 53.7, 63, 87.7, 100 and 120 kDa. Skin tests showed a delayed hypersensitivity. Immunoblotting of A. altissima pollen extract with pooled subject’s sera detected a major IgE-binding component of 42 kDa. Moreover, these results will provide a platform for cloning cDNA encoding allergenic protein from A. altissima pollen.     

Analysis of the pollen allergen content of twelve olive cultivars grown in Portugal

Olive pollen presents intercultivar variability as regards to its antigenic and allergenic composition. In this study, we report the presence of differences among the SDS-PAGE pollen protein profiles of twelve Portuguese olive cultivars. Though most soluble proteins from these extracts seemed similar, three bands of about 18, 20 and 22 kDa presented sharp differences in intensity among the cultivars analyzed. The dissimilarity of patient’s sera reactivity to these protein extracts and the presence of several allergens already characterized (Ole e 1, Ole e 2, Ole e 5 and Ole e 9) in the extracts were also investigated. Epidemiological data indicated that 53.3 % out of the 428 patients analyzed with reactivity to pollen extracts, presented specific IgE levels to Olea europaea. A representative number of these sera were assayed in immunoblotting experiments. The cultivars ‘Galega’ and ‘Conserva de Elvas’ displayed low reactivity to the sera of atopic patients, whereas the extracts corresponding to the cultivars ‘Cobrançosa’, ‘Ascolana’ and ‘Verdeal de Serpa’ led to higher IgE reactivity. The use of antibodies to the allergens Ole e 1, Ole e 2, Ole e 5 and Ole e 9 in immunoblotting experiments also allowed cultivar discrimination. The cultivar ‘Verdeal de Serpa’ presented the highest Ole e 1, Ole e 5 and Ole e 9 allergen loads but the lowest Ole e 2. ‘Carrasquenha’ was the second cultivar in terms of the higher allergen content. Oppositely, the lowest allergen loads were those of the cultivars ‘Galega’ and ‘Conserva de Elvas’ coincidentally with their low IgE reactivity. These data may help interpret physiological differences in pollen performance for successful olive fertilization and, moreover, to better define future strategies for allergy diagnosis and treatment by specific immunotherapy.     

Distribution pattern of allergenic plants in the Beijing metropolitan region

Urbanization has significantly affected the composition and distribution pattern of plant species within and around cities. Plants with airborne pollens can cause seasonal allergic symptoms that are intensified by increasing air pollution and temperature. In urban landscapes, the reduced native biodiversity, increased exotic biodiversity, and species homogenization may all affect the abundance and distribution of allergenic plants. We investigated the plants with airborne pollens in the Beijing metropolitan region to determine the distribution pattern of allergenic plants as influenced by urbanization. Our results show that native allergenic plants dominated the urban area in the region. The species richness of allergenic plants, particularly the exotic plants with airborne pollens, significantly differed between land use types. The higher the plant diversity in the urban area, the higher the frequency of allergenic plant occurrence. Despite the homogenization of the allergenic plant communities, their characteristic species were still present across the metropolitan region. The flowering allergenic plants also differed between the different land use types. We suggest that some common allergenic plants should be avoided by urban planners, especially those that are exotic to the region. Humans susceptible to pollinosis should stay away from places that are concentrated with allergenic plants, and be aware of the flowering phenology of allergenic plants.     

Size distribution of allergenic Cry j 2 released from airborne <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> pollen grains during the pollen scattering seasons

Japanese cedar (Cryptomeria japonica) pollinosis is one of seasonal allergic rhinitis that mainly occurs in Japan. The pollinosis is caused by two main kinds of allergenic proteins called Cry j 1 and Cry j 2 which exist in Cryptomeria japonica pollen. In our previous study, we reported that the size-segregated of airborne fine allergenic Cry j 1 and morphological change of Cry j 1 due to the contact with rainfall. However, the study on airborne allergenic Cry j 2 in different particle sizes has not been identified until now. Therefore, the main aim of this study is to investigate the size distribution and scattering behavior of allergenic Cry j 2. The Cry j 2 particles were collected and determined in different particle sizes at the urban sampling points during the most severe pollination season of 2012 in Saitama, Japan. After the size-segregated Cry j 2 allergenic particles were collected using an Andersen high-volume (AHV) atmospheric sample, the airborne Cry j 2 concentrations were determined with a surface plasmon resonance (SPR) method. At the same time, the airborne Cryptomeria japonica pollens were also counted by the Durham pollen sampler. The higher concentrations of the allergenic Cry j 2 were detected even in particle sizes equal to or less than 1.1 μm (PM_1.1) than other particle sizes. The airborne particles ranges from 0.06 to 11 μm were also collected by a low-pressure impactor (LPI) atmospheric sampler. After that, the concentrations of Cry j 2 allergenic particles in fine particle sizes were measured by the SPR method either. With the help of this study, we have confirmed the existence of fine daughter allergenic particles, which clearly differ from the parent pollen grains in size, especially after the rainy days. It is possible that the daughter allergenic species will be released from the fractions of cell wall and burst pollen grains. We concluded that rainwater was one of the important factors that affects the release of pollen allergenic proteins of both Cry j 1 and Cry j 2 from the parent pollen grains.     

In‐Depth Proteome Analysis of Ricinus communis Pollens

Pollen grains are tiny structures vital for sexual reproduction and consequently seed and fruit production in angiosperms, and a source of many allergenic components responsible for deleterious implications for health worldwide. Current pollen research is mainly focused on unraveling the molecular mechanisms underlying the pollen germination and tube formation passing from the quiescent stage. In this context, an in‐depth proteome analysis of the pollens from Ricinus communis at three different stages—that is, mature, hydrated, and in vitro germinated—is performed. This analysis results in the identification of 1950 proteins, including 1773, 1313, and 858, from mature, hydrated, and germinated pollens, respectively. Based on label‐free quantification, 164 proteins are found to be significantly differentially abundant from mature to hydrated pollens, 40 proteins from hydrated to germinated, and 57 proteins from mature to germinated pollens, respectively. Most of the differentially abundant proteins are related to protein, carbohydrate, and energy metabolism and signaling. Besides other functional classes, a reasonable number of the proteins are predicted to be allergenic proteins, previously undiscovered. This is the first in‐deep proteome analysis of the R. communis pollens and, to the best of our knowledge, one of the most complete proteome dataset identified from the pollens of any plant species, thus providing a reference proteome for researchers interested in pollen biology.