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Peach profilin: cloning, heterologous expression and cross‐reactivity with Bet v 2
Author(s) -
RodríguezPerez R.,
FernándezRivas M.,
GonzálezMancebo E.,
SánchezMonge R.,
DíazPerales A.,
Salcedo G.
Publication year - 2003
Publication title -
allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.363
H-Index - 173
eISSN - 1398-9995
pISSN - 0105-4538
DOI - 10.1034/j.1398-9995.2003.00198.x
Subject(s) - profilin , polyclonal antibodies , allergen , biology , cross reactivity , recombinant dna , microbiology and biotechnology , complementary dna , immunoglobulin e , allergy , antibody , biochemistry , gene , genetics , immunology , cross reactions , cytoskeleton , actin cytoskeleton , cell
Background: Peach is among the main foods causing allergic reactions in the Mediterranean adult population. Only a single peach allergen, named Pru p 3, has been characterized. However, a potential role of profilin has also been suggested in grass pollen‐associated allergy to peach. Methods: Complementary DNA clones for two different peach profilin isoforms were obtained by reverse transcriptase polymerase chain reaction using non‐degenerated primers. Expression of recombinant peach profilin was performed in Escherichia coli , and confirmed using rabbit polyclonal antibodies to sunflower pollen profilin. Twenty‐nine individual sera from patients with peach allergy proved by double‐blind, placebo‐controlled food challenges (DBPCFC), either with ( n = 15) or without ( n = 14) specific IgE to Bet v 2, were used in immunodetection assays to test recombinant peach profilin reactivity. Results: Each peach profilin cDNA included an open reading frame coding for a 131 amino acid protein. The peach profilin isoforms, designated Pru p 4.01 and Pru p 4.02, showed 80% of amino acid sequence identity, and were very similar (>70% identity) to allergenic profilins from plant foods and pollens. Recombinant Pru p 4.01 was expressed in E. coli as a nonfusion protein, displaying the expected molecular size and reacting with anti‐profilin antibodies. rPru p 4.01 was recognized by all sera (15 of 15) with specific IgE to Bet v 2, whereas no sera (zero of 14) without IgE to this birch allergen reacted with rPru p 4.01. Conclusions: Peach profilin Pru p 4 is very closed to other allergenic profilins from plant foods and pollens. A complete correlation between reactivity to rPru p 4 and rBet v 2 has been found in sera from peach allergic patients.