title: Proteomic analysis of major and minor allergens from isolated pollen cytoplasmic granules.
abstract: Grass pollen is one of the most important vectors of aeroallergens. Under atmospheric conditions, pollen grains can release pollen cytoplasmic granules (PCGs). The allergens associated with these intrinsic subfractions induce, in laboratory animals as well as in asthmatic patients, allergic and inflammatory responses. The objectives of this study were to characterize the PCGs' intrinsic allergens and to compare them with those of pollen grains. The water-soluble proteins were extracted from pollen grains and their PCGs. IgE-binding proteins were analyzed and characterized through an allergomic strategy: 1- and 2-dimensional gel electrophoresis (1-DE and 2-DE), immunoblotting, using grass-pollen-sensitized patient sera, mass spectrometry (MS) analysis, and database searching. Several of the allergens listed in the IUIS nomenclature, Phl p 1, 4, 5, 6, and 12, were detected in pollen and PCG extracts, whereas Phl p 11 was found only in PCGs, and Phl p 2 as well as Phl p 13 were found only in pollen extract. Some other allergens not listed in the IUIS nomenclature were also characterized in both pollen and PCG extracts. Since the major grass pollen allergens were found in PCGs and because of their small size, these submicronic particles should be considered as very potent sensitizing and challenging respirable vectors of allergens.
authors.affiliation: ESPCI ParisTech, UMR 7195 CNRS , LSABM, 10 rue Vauquelin, 75231 Paris Cedex 05, France. Oussama.Abou-chakra@espci.fr
authors.forename: Oussama R
authors.initials: OR
authors.lastname: Abou Chakra
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Sutra
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Demey Thomas
authors.forename: Joëlle
authors.initials: J
authors.lastname: Vinh
authors.forename: Ghislaine
authors.initials: G
authors.lastname: Lacroix
authors.forename: Pascal
authors.initials: P
authors.lastname: Poncet
authors.forename: Hélène
authors.initials: H
authors.lastname: Sénéchal
mesh_terms: D003593:Cytoplasm / Q000737:chemistry*; D031745:Dactylis; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000276:immunology; D013058:Mass Spectrometry; D010940:Plant Proteins / Q000032:analysis* / Q000276:immunology; D011058:Pollen / Q000737:chemistry* / Q000276:immunology; D020543:Proteome / Q000032:analysis* / Q000276:immunology; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology
chemical_list: D010940:Plant Proteins; D020543:Proteome; D007073:Immunoglobulin E