Biochemical composition of Phleum pratense pollen grains: A review.

The Poaceae family is composed of 12,000 plant species. Some of these species produce highly allergenic anemophilous pollen grains (PGs). Phleum pratense pollen grains (PPPGs) emerged as a model for studies related to grass allergy. The biochemical composition of allergenic PGs has not yet been fully described despite potential health effects of PG constituents other than allergenic proteins. This review brings together the information available in literature aiming at creating a comprehensive picture of the current knowledge about the chemical composition of allergenic PGs from timothy grass. PPPGs have an average diameter between 30-35 µm and the mass of a single PG was reported between 11 and 26 ng. The pollen cytoplasm is filled with two types of pollen cytoplasmic granules (PCGs): the starch granules and the polysaccharide particles (p-particles). Starch granules have a size between 0.6-2.5 µm with an average diameter of 1.1 µm (estimated number of 1000 granules per PG) while p-particles have a size ranging around 0.3 to 0.4 µm (estimated number between 61,000-230,000 p-particles per PG). The rupture of PG induces the release of PCGs and the dispersion of allergens in the inhalable fraction of atmospheric aerosol. PPPGs are composed of sporopollenin, sugars, polysaccharides, starch, glycoproteins (including allergens), amino-acids, lipids, flavonoids (including isorhamnetin), various elements (the more abundant being Si, Mg and Ca), phenolic compounds, phytoprostanoids, carotenoids (pigments) metals and adsorbed pollutants. PPPG contains about a hundred different proteins with molecular masses ranging from 10 to 94 kDa, with isoelectric points from 3.5-10.6. Among these proteins, allergens are classified in eleven groups from 1 to 13 with allergens from groups 1 and 5 being the major contributors to Phl p pollen allergy. Major allergen Phl p 5 was quantified in PPPGs by several studies with concentration ranging from 2.7 and 3.5 µg.mg-1 in unpolluted environment. Values for other allergens are scarce in literature; only one quantitative assessment exists for allergen groups Phl p 1, 2 and 4. The extractible lipid fraction of PPPGs is estimated between 1.7-2.2% of the total PG mass. The main chemical families of lipids reported in PPPGs are: alkanes, alkenes, alcohols, saturated and unsaturated fatty acids, di- and tri-hydroxylated fatty acids, aldehydes and sterols. Several lipid compounds with potential adjuvant effects on allergy have been specifically quantified in PPPGs: E2-like prostaglandin (PGE2), B4-like leukotriene (LTB4), unsaturated fatty acids (linoleic and linolenic acids and their hydroxylated derivatives), adenosine, vitamins and phenolic compounds. Some other biochemical characteristics such as NAD(P)H oxidase, protease activity and pollen microbiome were described in the literature. The bioaccessibility in physiological conditions has not been described for most biochemicals transported by allergenic PPPGs. There is also a considerable lack of knowledge about the potential health effects of pollen constituents other than allergens. The variability of pollen composition remains also largely unknown despite its importance for plant reproduction and allergy in an environment characterized by chemical pollution, climate change and loss of biodiversity.

Organic and aqueous extraction of lipids from birch pollen grains exposed to gaseous pollutants.

The lipid fraction of birch pollen grains (BPGs) is not yet fully described, although pollen lipid molecules may play a role in the allergic immune response. The mechanisms by which atmospheric pollutants modify allergenic pollen grains (PGs) are also far from being elucidated despite high potential effects on allergic sensitization. This work is a contribution to a better description of the lipid profile (both external and cytoplasmic) of BPGs and of alterations induced by gaseous air pollutants. Several lipid extractions were performed using organic and aqueous solvents on BPGs following exposure to ozone and/or nitrogen dioxide and under conditions favoring the release of internal lipids. Ozone reacted with alkenes to produce aldehydes and saturated fatty acids, while nitrogen dioxide was shown to be unreactive with lipids. NO2 exhibited a protective effect against the reactivity of alkenes with ozone, probably by competition for adsorption sites. The decreased reactivity of ozone during simultaneous exposure to NO2/O3 raised the possibility of a Langmuir-Hinshelwood mechanism. Oxidation reactions induced by exposure of BPGs to ozone did not substantially modify the extraction of lipids by aqueous solvent, suggesting that the bioaccessibility of lipids was not modified by oxidation. On the contrary, the rupture of PGs appeared to be a key factor in enhancing the bioaccessibility of bioactive lipid mediators (linoleic and α-linolenic acids) in an aqueous solution. The internal lipid fraction of BPGs has specific characteristics compared with external lipids, with more abundant hexadecanoic acid, tricosanol, and particularly unsaturated fatty acids (linoleic and α-linolenic acids). Several mechanisms of action of gaseous pollutants on allergenic pollen were identified in this study: gaseous air pollutants can (i) modify the external lipid fraction by reactivity of alkenes, (ii) adsorb on the surface of PGs and be a source of oxidative stress after inhalation of PGs, and (iii) promote the release of cytoplasmic bioactive lipids by facilitating pollen rupture.

Uptake of ozone and modification of lipids in Betula Pendula pollen.

Pollen allergy risk is modified by air pollutants, including ozone, but the chemical modifications induced on pollen grains are poorly understood. Pollen lipidic extract has been shown to act as an adjuvant to the allergenic reaction and therefore, the modification of lipids by air pollutants could have health implications. Birch pollen was exposed in vitro to ozone to explore the reactivity of O3 on its surface and on its lipidic fraction. Uptake coefficients of ozone were determined for ozone concentration of 117 ppb on the surface of native birch pollen (8.6 ±â€¯0.8 × 10-6), defatted pollen (9.9 ±â€¯0.9 × 10-6), and for crushed pollen grains (34±3 × 10-6). The mass of ozone uptaken was increased by a factor of four for crushed pollen compared to native pollen showing a higher susceptibility to ozone of cytoplasmic granules and broken pollen grains. A total mass of extractible lipids of 27 mg per gram of birch pollen was found and a fraction of these lipids was identified and quantified (fatty acids, alkanes, alkenes and aldehydes). The distribution of lipids was modified by ozone exposure of 115 and 1000 ppb for 16 h with the following reactivity: consumption of alkene, formation of aldehydes and formation of nonanoic acid and octadecanoic acid. The quantity of ozone trapped in the lipidic fraction during 15 min at 115 ppb is enough to contribute to the reactivity of one-third of the alkenes demonstrating that pollen could be susceptible to an atmospheric increase of ozone concentration even for a very short duration complicating the understanding of the link between pollen allergy and pollution.

Chemical modification of coating of Pinus halepensis pollen by ozone exposure.

Pollen coating, located on the exine, includes an extractible lipid fraction. The modification of the pollen coating by air pollutants may have implications on the interactions of pollen with plant stigmas and human cells. Pinus halepensis pollen was exposed to ozone in vitro and the pollen coating was extracted with organic solvent and analyzed by GC-MS. Ozone has induced chemical changes in the coating as observed with an increase in dicarboxylic acids, short-chain fatty acids and aldehydes. 4-Hydroxybenzaldehyde was identified as the main reaction product and its formation was shown to occur both on native pollen and on defatted pollen. 4-Hydroxybenzaldehyde is very likely formed via the ozonolysis of acid coumaric-like monomers constitutive of the sporopollenin. Modification of pollen coating by air pollutants should be accounted for in further studies on effect of pollution on germination and on allergenicity.