A human monoclonal IgE antibody defines a highly allergenic fragment of the major timothy grass pollen allergen, Phl p 5: Molecular, immunological, and structural characterization of the epitope-containing domain

S. Flicker, S. Vrtala, P. Steinberger, L. Vangelista, A. Bufe, A. Petersen, M. Ghannadan, W. R. Sperr, P. Valent, L. Norderhaug, B. Bohle, H. Stockinger, C. Suphioglu, Kok Ong Eng Kok Ong, D. Kraft, R. Valenta

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Almost 90% of grass pollen-allergic patients are sensitized against group 5 grass pollen allergens. We isolated a monoclonal human IgE Fab out of a combinatorial library prepared from lymphocytes of a grass pollen-allergic patient and studied its interaction with group 5 allergens. The IgE Fab cross-reacted with group 5A isoallergens from several grass and corn species. By allergen gene fragmentation we mapped the binding site of the IgE Fab to a 11.2-kDa N-terminal fragment of the major timothy grass pollen allergen Pld p 5A. The IgE Fab-defined Phl p 5A fragment was expressed in Escherichia coli and purified to homogeneity. Circular dichroism analysis revealed that the rPhl p 5A domain, as well as complete rPld p 5A, assumed a folded conformation consisting predominantly of an a helical secondary structure, and exhibited a remarkable refolding capacity. It reacted with serum IgE from 76% of grass pollen-allergic patients and revealed an extremely high allergenic activity in basophil histamine release as well as skin test experiments. Thus, the rPhl p 5A domain represents an important allergen domain containing several IgE epitopes in a configuration optimal for efficient effector cell activation. We suggest the rPhl p 5A fragment and the corresponding IgE Fab as paradigmatic tools to explore the structural requirements for highly efficient effector cell activation and, perhaps later, for the development of generally applicable allergen-specific therapy strategies.

Original languageEnglish
Pages (from-to)3849-3859
Number of pages11
JournalJournal of Immunology
Volume165
Issue number7
Publication statusPublished - Oct 1 2000
Externally publishedYes

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Phleum
Pollen
Allergens
Immunoglobulin E
Epitopes
Monoclonal Antibodies
Poaceae
Basophils
Histamine Release
Circular Dichroism
Skin Tests
Zea mays
Binding Sites
Lymphocytes
Escherichia coli

ASJC Scopus subject areas

  • Immunology

Cite this

A human monoclonal IgE antibody defines a highly allergenic fragment of the major timothy grass pollen allergen, Phl p 5 : Molecular, immunological, and structural characterization of the epitope-containing domain. / Flicker, S.; Vrtala, S.; Steinberger, P.; Vangelista, L.; Bufe, A.; Petersen, A.; Ghannadan, M.; Sperr, W. R.; Valent, P.; Norderhaug, L.; Bohle, B.; Stockinger, H.; Suphioglu, C.; Eng Kok Ong, Kok Ong; Kraft, D.; Valenta, R.

In: Journal of Immunology, Vol. 165, No. 7, 01.10.2000, p. 3849-3859.

Research output: Contribution to journalArticle

Flicker, S, Vrtala, S, Steinberger, P, Vangelista, L, Bufe, A, Petersen, A, Ghannadan, M, Sperr, WR, Valent, P, Norderhaug, L, Bohle, B, Stockinger, H, Suphioglu, C, Eng Kok Ong, KO, Kraft, D & Valenta, R 2000, 'A human monoclonal IgE antibody defines a highly allergenic fragment of the major timothy grass pollen allergen, Phl p 5: Molecular, immunological, and structural characterization of the epitope-containing domain', Journal of Immunology, vol. 165, no. 7, pp. 3849-3859.
Flicker, S. ; Vrtala, S. ; Steinberger, P. ; Vangelista, L. ; Bufe, A. ; Petersen, A. ; Ghannadan, M. ; Sperr, W. R. ; Valent, P. ; Norderhaug, L. ; Bohle, B. ; Stockinger, H. ; Suphioglu, C. ; Eng Kok Ong, Kok Ong ; Kraft, D. ; Valenta, R. / A human monoclonal IgE antibody defines a highly allergenic fragment of the major timothy grass pollen allergen, Phl p 5 : Molecular, immunological, and structural characterization of the epitope-containing domain. In: Journal of Immunology. 2000 ; Vol. 165, No. 7. pp. 3849-3859.
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abstract = "Almost 90{\%} of grass pollen-allergic patients are sensitized against group 5 grass pollen allergens. We isolated a monoclonal human IgE Fab out of a combinatorial library prepared from lymphocytes of a grass pollen-allergic patient and studied its interaction with group 5 allergens. The IgE Fab cross-reacted with group 5A isoallergens from several grass and corn species. By allergen gene fragmentation we mapped the binding site of the IgE Fab to a 11.2-kDa N-terminal fragment of the major timothy grass pollen allergen Pld p 5A. The IgE Fab-defined Phl p 5A fragment was expressed in Escherichia coli and purified to homogeneity. Circular dichroism analysis revealed that the rPhl p 5A domain, as well as complete rPld p 5A, assumed a folded conformation consisting predominantly of an a helical secondary structure, and exhibited a remarkable refolding capacity. It reacted with serum IgE from 76{\%} of grass pollen-allergic patients and revealed an extremely high allergenic activity in basophil histamine release as well as skin test experiments. Thus, the rPhl p 5A domain represents an important allergen domain containing several IgE epitopes in a configuration optimal for efficient effector cell activation. We suggest the rPhl p 5A fragment and the corresponding IgE Fab as paradigmatic tools to explore the structural requirements for highly efficient effector cell activation and, perhaps later, for the development of generally applicable allergen-specific therapy strategies.",
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AU - Flicker, S.

AU - Vrtala, S.

AU - Steinberger, P.

AU - Vangelista, L.

AU - Bufe, A.

AU - Petersen, A.

AU - Ghannadan, M.

AU - Sperr, W. R.

AU - Valent, P.

AU - Norderhaug, L.

AU - Bohle, B.

AU - Stockinger, H.

AU - Suphioglu, C.

AU - Eng Kok Ong, Kok Ong

AU - Kraft, D.

AU - Valenta, R.

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