Cloning and characterization of profilin (Pru du 4), a cross-reactive almond (Prunus dulcis) allergen

Pallavi Tawde, Yeldur P. Venkatesh, Fang Wang, Suzanne S Teuber, Shridhar K. Sathe, Kenneth H. Roux

Research output: Contribution to journalArticle

52 Scopus citations

Abstract

Background: The identity of allergenic almond proteins is incomplete. Objective: Our objective was to characterize patient IgE reactivity to a recombinant and corresponding native almond allergen. Methods: An almond cDNA library was screened with sera from patients with allergy for IgE binding proteins. Two reactive clones were sequenced, and 1 was expressed. The expressed recombinant allergen and its native counterpart (purified from unprocessed almond flour) were assayed by 1-dimensional and 2-dimensional gel electrophoresis, dot blot, and ELISA, and screened for cross-reactivity with grass profilin. Results: The 2 selected clones encoded profilin (designated Pru du 4) sequences that differed by 2 silent mutations. By dot-blot analyses, 6 of 18 patient sera (33%) reacted with the recombinant Pru du 4 protein, and 8 of 18 (44%) reacted with the native form. ELISA results were similar. Almond and ryegrass profilins were mutually inhibitable. Two-dimensional immunoblotting revealed the presence of more than 1 native almond profilin isoform. The strength of reactivity of some patients' serum IgE differed markedly between assays and between native and recombinant profilins. Conclusion: Almond nut profilin is an IgE-binding food protein that is cross-reactive with grass pollen profilin and is susceptible to denaturation, resulting in variable reactivity between assay types and between patients. Clinical implications: Serum IgE of nearly half of the tested patients with almond allergy reacts with almond nut profilin. Because most patients also had pollinosis, the well-known cross-reactivity between pollen and food profilins could account for this pattern of reactivity.

Original languageEnglish (US)
Pages (from-to)915-922
Number of pages8
JournalJournal of Allergy and Clinical Immunology
Volume118
Issue number4
DOIs
StatePublished - Oct 2006

Keywords

  • Allergen
  • almond allergy
  • food allergy
  • pan-allergen
  • profilin
  • recombinant allergen

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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