BACKGROUND: Wheat protein and its derivatives can cause protein contact dermatitis (PCD), which mainly occurs in bakers. Few studies have attempted to identify the allergens responsible for wheat PCD. OBJECTIVES: The aim of this study was to identify allergenic wheat proteins in patients with wheat PCD. METHODS: Water-soluble and water-insoluble wheat flour proteins were separated by 1- or 2-dimensional gel electrophoresis. IgE-binding proteins were detected by immunoblotting with sera from 3 wheat PCD patients and identified by N-terminal amino acid sequence analysis. The IgE-binding proteins were recombinantly expressed in Escherichia coli and tested against patients' sera. RESULTS: IgE antibodies from the patients' sera reacted with water-soluble proteins rather than water-insoluble proteins, and the 2-dimensional electrophoresis and immunoblotting produced individual IgE-binding patterns. Analysis of the N-terminal amino acid sequence of the IgE-binding proteins from the 2-dimensional gel led to the identification of three glycoproteins, wheat 27-kDa allergen, peroxidase, and purple acid phosphatase. No specific IgE antibodies to their non-glycosylated recombinant proteins were observed. CONCLUSIONS: We identified wheat 27-kDa allergen, peroxidase and purple acid phosphatase as candidate allergens for wheat PCD. Our results suggest that glycan moieties in these proteins are involved in IgE binding.
BACKGROUND:Wheat protein and its derivatives can cause protein contact dermatitis (PCD), which mainly occurs in bakers. Few studies have attempted to identify the allergens responsible for wheatPCD. OBJECTIVES: The aim of this study was to identify allergenic wheat proteins in patients with wheatPCD. METHODS:Water-soluble and water-insoluble wheat flour proteins were separated by 1- or 2-dimensional gel electrophoresis. IgE-binding proteins were detected by immunoblotting with sera from 3 wheatPCDpatients and identified by N-terminal amino acid sequence analysis. The IgE-binding proteins were recombinantly expressed in Escherichia coli and tested against patients' sera. RESULTS: IgE antibodies from the patients' sera reacted with water-soluble proteins rather than water-insoluble proteins, and the 2-dimensional electrophoresis and immunoblotting produced individual IgE-binding patterns. Analysis of the N-terminal amino acid sequence of the IgE-binding proteins from the 2-dimensional gel led to the identification of three glycoproteins, wheat 27-kDa allergen, peroxidase, and purple acid phosphatase. No specific IgE antibodies to their non-glycosylated recombinant proteins were observed. CONCLUSIONS: We identified wheat 27-kDa allergen, peroxidase and purple acid phosphatase as candidate allergens for wheatPCD. Our results suggest that glycan moieties in these proteins are involved in IgE binding.
Authors: Carla Viegas; Gerard T A Fleming; Abdul Kadir; Beatriz Almeida; Liliana Aranha Caetano; Anita Quintal Gomes; Magdalena Twarużek; Robert Kosicki; Susana Viegas; Ann Marie Coggins Journal: Microorganisms Date: 2020-01-15
Authors: Margitta Worm; Imke Reese; Barbara Ballmer-Weber; Kirsten Beyer; Stephan C Bischoff; Martin Classen; Peter J Fischer; Thomas Fuchs; Isidor Huttegger; Uta Jappe; Ludger Klimek; Berthold Koletzko; Lars Lange; Ute Lepp; Vera Mahler; Bodo Niggemann; Ute Rabe; Martin Raithel; Joachim Saloga; Christiane Schäfer; Sabine Schnadt; Jens Schreiber; Zsolt Szépfalusi; Regina Treudler; Martin Wagenmann; Bernhard Watzl; Thomas Werfel; Torsten Zuberbier; Jörg Kleine-Tebbe Journal: Allergo J Int Date: 2015-11-07