Food and Drug Reactions and Anaphylaxis
Identification of wheat flour allergens by means of 2-dimensional immunoblotting

https://doi.org/10.1067/mai.2001.113761Get rights and content

Abstract

Background: Wheat flour proteins are allergens for 60% to 70% of bakers with workplace-related respiratory symptoms. Objective: The aim of the study was to investigate the variability of IgE antibody patterns of wheat flour–sensitized bakers and to identify the most frequently recognized allergens. Methods: Water/salt-soluble wheat flour proteins from the cultivar Bussard were separated by using 2-dimensional gel electrophoresis with immobilized pH gradients. IgE-reactive proteins were identified by means of immunoblotting with sera of 10 subjects with baker’s asthma. Mass spectrometric fingerprinting was used to identify the proteins most frequently recognized by IgE. Results: The IgE immunoblots obtained with 10 different sera exhibited a remarkable heterogeneity. Each patient showed an individual IgE-binding pattern with 4 to 50 different allergen spots. Altogether, more than 100 IgE-binding protein spots were detected. Nine of the predominant IgE-binding protein spots were identified by using mass spectrometric fingerprinting. The obtained masses matched 2 different isoforms of glycerinaldehyde-3-phosphate dehydrogenase from Hordeum vulgare , triosephosphate isomerase from H vulgare , and serpin, a serine proteinase inhibitor from Triticum aestivum . Conclusions: The results show a great interindividual variation of IgE-binding patterns of wheat flour proteins in baker’s asthma. The clinical relevance of the identified 4 new allergens will be further investigated in the near future. (J Allergy Clin Immunol 2001;107:907-13.)

Section snippets

Baker’s sera and specific IgE measurement

Sera were obtained from 10 bakers with bronchial asthma, rhinitis, or both caused by type I allergy to flour. Severity of asthma symptoms was classified according to the “Guidelines for the diagnosis and management of asthma.”15 As controls, one serum from a baker without clinical symptoms and without specific IgE to any of the tested baker allergens and one serum of a patient who had never worked in a bakery were used. All patients were men.

Human IgE antibodies to wheat flour, rye flour, soy

Description of patients

Ten patients with work-related wheat flour allergy were recruited for the identification of wheat grain albumins and globulins bound by IgE antibodies. All patients had workplace-related asthma and high levels of specific IgE to wheat and rye flour (Table I). Additional sensitizations to soy flour, house dust mites, enzymes used for the baking process, and grass pollens were found. The control patients had no specific IgE to wheat or rye flour.

Two-dimensional electrophoresis of wheat flour proteins

Water/salt-soluble wheat grain proteins of the

Discussion

Water/salt-soluble wheat flour proteins of the cultivar Bussard were analyzed by using high-resolution 2-dimensional electrophoresis with IPGs. Similar to the results of Posch et al,7 who investigated the cultivar Fregatt with the same method, we obtained a complex but highly reproducible pattern of several hundred silver-stained albumin and globulin spots. The high reproducibility of spot positions in 2-dimensional electrophoresis with IPGs offers the possibility of producing a series of

Acknowledgements

We thank Maria Düser and Silke Maryska for excellent technical assistance, Bernd Naurath and Markus Bocian for photographs and scans, and Karin Weber for assistance in improving the English.

References (23)

  • R Sánchez-Monge et al.

    Wheat and barley allergens associated with baker’s asthma. Glycosylated subunits of the alpha-amylase-inhibitor family have enhanced IgE-binding capacity

    Biochem J

    (1992)
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