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| Article Authors: | Ulrike Schulmeister; Heidrun Hochwallner; Ines Swoboda; Margarete Focke-Tejkl; Beate Geller; Mats Nystrand; Annika Härlin; Josef Thalhamer; Sandra Scheiblhofer; Walter Keller; Bodo Niggemann; Santiago Quirce; Christoph Ebner; Adriano Mari; Gabrielle Pauli; Udo Herz; Rudolf Valenta; Susanne Spitzauer |
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| Article Title: | Cloning, expression, and mapping of allergenic determinants of alphaS1-casein, a major cow's milk allergen. |
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| Reference ID: | 1014862 |
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| Abstract: | Milk is one of the first components introduced into human diet. It also represents one of the first allergen sources, which induces IgE-mediated allergies in childhood ranging from gastrointestinal, skin, and respiratory manifestations to severe life-threatening manifestations, such as anaphylaxis. Here we isolated a cDNA coding for a major cow's milk allergen, alphaS1-casein, from a bovine mammary gland cDNA library with allergic patients' IgE Abs. Recombinant alphaS1-casein was expressed in Escherichia coli, purified, and characterized by circular dichroism as a folded protein. IgE epitopes of alphaS1-casein were determined with recombinant fragments and synthetic peptides spanning the alphaS1-casein sequence using microarrayed components and sera from 66 cow's milk-sensitized patients. The allergenic activity of ralphaS1-casein and the alphaS1-casein-derived peptides was determined using rat basophil leukemia cells transfected with human FcepsilonRI, which had been loaded with the patients' serum IgE. Our results demonstrate that ralphaS1-casein as well as alphaS1-casein-derived peptides exhibit IgE reactivity, but mainly the intact ralphaS1-casein induced strong basophil degranulation. These results suggest that primarily intact alphaS1-casein or larger IgE-reactive portions thereof are responsible for IgE-mediated symptoms of food allergy. Recombinant alphaS1-casein as well as alphaS1-casein-derived peptides may be used in clinical studies to further explore pathomechanisms of food allergy as well as for the development of new diagnostic and therapeutic strategies for milk allergy. |
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| Affiliations: | Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Vienna, Austria. |
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| Date: | 2009 |
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| Reference Type: | Literature |
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| PubMed ID: | 19454699 |
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| Journal: | J Immunol |
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| Journal Volume: | 182 |
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| Article Pages: | 7019-29 |
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| Journal ISSN: | 1550-6606 |
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| Article Chemical List: | Allergens;Caseins;DNA, Complementary;Epitopes;Peptide Fragments;Receptors, IgE;Immunoglobulin E |
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| Article MeSH List: | Allergens(immunology); Animals; Basophils(physiology); Caseins(immunology); Cattle; Cell Degranulation; Cell Line, Tumor; Cloning, Molecular; DNA, Complementary; Epitope Mapping; Epitopes(genetics; immunology); Humans; Immunoglobulin E; Milk(immunology); Milk Hypersensitivity(immunology); Peptide Fragments(chemical synthesis; immunology); Rats; Receptors, IgE |
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| Curation Last Updated: | 2013-05-28 21:43:59 |
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