Mapping of IgE-binding epitopes on the recombinant major group I allergen of velvet grass pollen, rHol I 1

被引:26
作者
Schramm, G
Bufe, A
Petersen, A
Haas, H
Schlaak, M
Becker, WM
机构
[1] Forschungszentrum Borstel, Division of Allergology, D-23845 Borstel
关键词
group I allergens; velvet grass pollen; recombinant allergens; B-cell epitope mapping;
D O I
10.1016/S0091-6749(97)80012-7
中图分类号
R392 [医学免疫学];
学科分类号
100102 ;
摘要
Background: New and more successful approaches to diagnosis and therapy of allergic diseases require a more subtle understanding of the structure and the epitopes on the allergen molecule. Objective: This study was done to obtain more information on the structure and the IgE-binding epitopes of a major allergen of velvet grass pollen, Hol I 1. Methods: We cloned Hot I 1 from a complementary DNA library and performed B-cell epitope mapping with 21 recombinant fragments expressed as fusion proteins in Escherichia coli. The fragments were analyzed by Western blotting with sera from 50 different patients. Results: The patients' sera individually recognized at least four different IgE-binding regions (amino acids 1 to 27, 61 to 76, 84 to 105, and 158 to 240). According to their binding patterns with these epitopes, they were divided into five groups. Most sera (92%) bound to the C-terminal peptide (158 to 240), which consists of more than 80 amino acids, whereas there was virtually no binding to smaller fragments covering this region. In contrast to the C-terminal peptide, the IgE-binding peptides on the N terminus and on the middle region of the molecule were of a smaller size (15 to 30 amino acids). Conclusions: The major group I allergen of velvet grass bears at least four different IgE-binding epitopes, which were individually recognized by sera from different patients. The C terminus represents the major IgE-binding region and contains at least one discontinuous IgE-binding epitope, whereas the N terminus and middle region of Hol I 1 seem to contain continuous IgE-binding epitopes.
引用
收藏
页码:781 / 787
页数:7
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