Cross-reactive and species-specific immunoglobulin E epitopes of plant profilins: an experimental and structure-based analysis.
BACKGROUND
Profilins are cross-reactive plant allergens responsible for multiple pollen sensitization and pollen-associated food allergy. While it is assumed that profilins from different species are immunologically equivalent, some studies suggest partial or even lacking IgE cross-reactivity between certain profilins.


OBJECTIVE
We aimed to obtain a semi-quantitative assessment of the contributions of conserved and species-specific epitopes to IgE binding of plant profilins.


METHODS
We compared model structures of profilins from timothy, mugwort, celery and bell pepper with crystal structures of birch and latex profilins. We predicted potential conformational epitopes that consisted of contiguous patches of at least 20% surface-exposed residues. Celery and timothy profilins were purified from their natural sources, and profilins from birch, mugwort, bell pepper and latex were expressed in Escherichia coli. The structural integrity of all purified proteins was confirmed by circular dichroism spectroscopy. IgE ELISAs and ELISA inhibitions using sera from 22 profilin-sensitized allergic patients were carried out.


RESULTS
Peptide backbone conformations of all six profilins were highly similar. Nine variable epitopes and two containing high proportions of conserved residues were predicted. IgE from all sera bound to all tested profilins and the amounts were highly correlated. However, IgE inhibition experiments revealed that up to 60% of total IgE binding was mediated by species-specific epitopes. The extent of cross-reactivity among profilins from timothy, birch, latex and celery was greater than cross-reactivity to mugwort and bell pepper profilins. This pattern was mirrored by sequence similarities among one of the predicted variable epitopes. Patients with IgE to cross-reactive epitopes displayed allergic reactions to a greater number of plant foods than patients having IgE directed to species-specific epitopes.


CONCLUSIONS
The large extent of cross-reactivity among plant profilins justifies using a single profilin for diagnosis. However, the fine specificity of IgE directed to variable epitopes may influence the clinical manifestation of profilin sensitization.
Radauer
C
C
Department of Pathophysiology, Center of Physiology and Pathophysiology, Medical University of Vienna, Vienna, Austria.
Willerroider
M
M
Fuchs
H
H
Hoffmann-Sommergruber
K
K
Thalhamer
J
J
Ferreira
F
F
Scheiner
O
O
Breiteneder
H
H
D000595:Amino Acid Sequence; D001667:Binding, Competitive; D003429:Cross Reactions; D000939:Epitopes / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D005512:Food Hypersensitivity / Q000276:immunology; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology*; D007073:Immunoglobulin E / Q000276:immunology*; D020315:Latex Hypersensitivity / Q000276:immunology; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology*; D011058:Pollen / Q000276:immunology; D051304:Profilins / Q000235:genetics / Q000276:immunology*; D017434:Protein Structure, Tertiary; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology; D016415:Sequence Alignment; D013045:Species Specificity
D000939:Epitopes; D010940:Plant Proteins; D051304:Profilins; D007073:Immunoglobulin E