Evaluation of ash pollen sensitization pattern using proteomic approach with individual sera from allergic patients.
BACKGROUND
In Europe, sensitization to ash pollen induces pollinosis with cross-reactivities with other pollen sources. The aim of the study was to identify the repertoire of ash pollen allergens and evaluate the extent of the diversity of the IgE response in ash allergic patients.


METHODS
The IgE reactivities of 114 ash pollen- and eight grass pollen-sensitized patients were screened by 1D immunoblot (SDS-PAGE) against ash pollen extract. The IgE reactivities of 13 ash pollen- and two grass pollen-sensitized patients were then evaluated in 2D immunoblots. Some IgE- and non-IgE-reactive proteins were identified by mass spectrometry.


RESULTS
In 1D analysis, 86% of sera showed binding to Fra e 1 (18-20 kDa), 23% to Fra e 2 (14 kDa), 3% to Fra e 3 (10 kDa) and 57% to High Molecular Weight allergens (HMW, >30 kDa). Individual analysis of 2D immunoblots showed several IgE-binding protein areas among which three were more often recognized: (i) Fra e 1 comprising, at least, 15 isoforms, (ii) a series of acidic spots (45 kDa), and (iii) Fra e 2, the ash profilin. HMW allergens could be resolved in four areas; two unidentified, one homologous to beta-galactosidase and the other to sugar transport proteins. A malate deshydrogenase and calmodulin were shown to be IgE-binding proteins and 10 non-IgE reactive proteins were identified.


CONCLUSIONS
No direct correlation was evidenced between IgE profile and the degree of sensitization even though 2 spectrotypes could be distinguished. Our data contribute to a better delineation of ash pollen allergens and patterns of sensitization.
Poncet
P
P
Ecole Supérieure de Physique et Chimie Industrielles de la Ville de Paris, CNRS, France.
Senechal
H
H
Clement
G
G
Purohit
A
A
Sutra
J-P
JP
Desvaux
F-X
FX
Wal
J-M
JM
Pauli
G
G
Peltre
G
G
Gougeon
M-L
ML
D015153:Blotting, Western; D003429:Cross Reactions / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D031661:Fraxinus / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood* / Q000276:immunology; D040901:Proteomics; D006255:Rhinitis, Allergic, Seasonal / Q000097:blood* / Q000453:epidemiology / Q000276:immunology*; D012882:Skin Tests; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D007073:Immunoglobulin E