A recombinant hypoallergenic parvalbumin mutant for immunotherapy of IgE-mediated fish allergy

Ines Swoboda, Agnes Bugajska-Schretter, Birgit Linhart, Petra Verdino, Walter Keller, Ulrike Schulmeister, Wolfgang R. Sperr, Peter Valent, Gabriel Peltre, Santiago Quirce, Nikolaos Douladiris, Nikolaos G. Papadopoulos, Rudolf Valenta, Susanne Spitzauer

    Research output: Contribution to journalArticlepeer-review

    Abstract

    IgE-mediated allergy to fish is a frequent cause of severe anaphylactic reactions. Parvalbumin, a small calcium-binding protein, is the major fish allergen. We have recently isolated a cDNA coding for carp parvalbumin, Cyp c 1, and expressed in Escherichia coli a recombinant Cyp c 1 molecule, which contained most IgE epitopes of saltwater and freshwater fish. In this study, we introduced mutations into the calcium-binding domains of carp parvalbumin by site-directed mutagenesis and produced in E. coli three parvalbumin mutants containing amino acid exchanges either in one (single mutants; Mut-CD and Mut-EF) or in both of the calcium-binding sites (double mutant; Mut-CD/EF). Circular dichroism analyses of the purified derivatives and the wild-type allergen showed that Mut-CD/EF exhibited the greatest reduction of overall protein fold. Dot blot assays and immunoblot inhibition experiments performed with sera from 21 fish-allergic patients showed that Mut-CD/EF had a 95% reduced IgE reactivity and represented the derivative with the least allergenic activity. The latter was confirmed by in vitro basophil histamine release assays and in vivo skin prick testing. The potential applicability for immunotherapy of Mut-CD/EF was demonstrated by the fact that mouse IgG Abs could be raised by immunization with the mutated molecule, which cross-reacted with parvalbumins from various fish species and inhibited the binding of fish-allergic patients' IgE to the wild-type allergen. Using the hypoallergenic carp parvalbumin mutant Mut-CD/EF, it may be possible to treat fish allergy by immunotherapy. Copyright © 2007 by The American Association of Immunologists, Inc.
    Original languageEnglish
    Pages (from-to)6290-6296
    Number of pages6
    JournalJournal of Immunology
    Volume178
    Issue number10
    Publication statusPublished - 15 May 2007

    Fingerprint

    Dive into the research topics of 'A recombinant hypoallergenic parvalbumin mutant for immunotherapy of IgE-mediated fish allergy'. Together they form a unique fingerprint.

    Cite this