Peach profilin:: cloning, heterologous expression and cross-reactivity with Bet v 2

被引:44
作者
Rodríguez-Perez, R
Fernández-Rivas, M
González-Mancebo, E
Sánchez-Monge, R
Díaz-Perales, A
Salcedo, G
机构
[1] ETS Ingn Agronomos, Dept Biotecnol, Unidad Bioquim, Madrid 28040, Spain
[2] Fdn Hosp Alcorcon, Unidad Alergia, Madrid, Spain
关键词
cross-reactivity; food allergy; peach; profilin; recombinant allergen;
D O I
10.1034/j.1398-9995.2003.00198.x
中图分类号
R392 [医学免疫学];
学科分类号
100102 ;
摘要
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.
引用
收藏
页码:635 / 640
页数:6
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