Food and Drug Reactions and AnaphylaxisMutational analysis of major, sequential IgE-binding epitopes in αs1-casein, a major cow's milk allergen☆,☆☆
Section snippets
Patient population
Sera from 15 patients with documented IgE-mediated cow's milk hypersensitivity (mean age, 8.5 years; range, 2-18 years) were used to identify the AAs essential for IgE binding. Milk-specific IgE antibodies in these sera ranged from 25 to >100 kU/L (median, >100 kU/L), as measured by the CAP system FEIA (Pharmacia Diagnostics, Uppsala, Sweden). In 11 children, CMA was confirmed by open (n = 3) or double-blind, placebo-controlled food challenges (n = 8). The remaining 4 children had a history of
Results
Pooled sera from 15 patients and individual sera from 8 randomly selected patients whose serum was included in the pool were used to identify AAs essential for epitope-specific recognition by IgE antibodies. In the first phase of experiments, each residue within an epitope was substituted with alanine (or, when alanine was present, with glycine) 1 at a time. Fig 1 shows an example of the immunolabeling with pooled sera for peptide AA 109-120.
Discussion
Within the last few years, the primary AA sequences of a number of food allergens and their IgE-binding regions have been identified. To develop safe immunotherapeutic agents for food allergy, modified, recombinant proteins need to be engineered that will not bind serum and mast cell IgE. Therefore, critical AAs within the IgE-binding regions need to be identified to provide the necessary information to alter the cDNAs that encode proteins capable of activating milk-specific T cells, but not
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Supported by grants AI 44236 and AI 24439 from the National Institute of Allergy and Infectious Diseases, grant M01 RR00071 from the Division of Research Resources, National Institutes of Health, and the Bunning Family Fund.
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Reprint requests: Kirsten Beyer, MD, Department of Pediatrics, Charite-CVK, Augustenburgerplatz 1, 13353 Berlin, Germany.