Cloning and Characterisation of Two IgE-Binding Proteins, Homologous to Tropomyosin and [alpha]-Tubulin, from the Mite Lepidoglyphus destructor

• Published in: International archives of allergy and immunology Vol. 130; no. 4; p. 258
• Main Authors: Saarne, Tiiu, Kaiser, Liselotte, Rasool, Omid, Huecas, Sonia, Marianne van Hage-Hamsten, Gafvelin, Guro
• Format: Journal Article
• Language: English
• Published: Basel S. Karger AG 01.04.2003
• ISSN: 1018-2438; 1423-0097

<Background:< The dust mite <Lepidoglyphus destructor< is a major source of mite allergy in European rural environments, but it also causes allergy in urban populations around the world. We have previously cloned, sequenced and expressed several allergens from <L. destructor< (Lep d 2, Lep d 5, Lep d 7 and Lep d 13). The aim of this study was to identify and clone additional allergens from <L. destructor<, and to evaluate their IgE-binding reactivities. <Methods:< PCR and screening with sera from <L. destructor<-sensitised individuals were used to isolate new clones from a phage display <L. destructor< cDNA library. The complete coding sequences of the clones were determined and expressed as His<6<-tagged recombinant proteins in <Escherichia coli<. The recombinant proteins were analysed by SDS-PAGE, immunoblotting and mass spectrometry. <Results:< Two new clones, showing homology to tropomyosin and [alpha]-tubulin in several species, were isolated from the phage display <L. destructor< cDNA library. Due to its homology to group 10 dust mite allergens, the tropomyosin clone was named Lep d 10. The IgE-binding frequencies of the recombinant Lep d 10 and [alpha]-tubulin were 13% (18/136) and 12% (11/95), respectively, among subjects with IgE reactivity to mites and/or crustaceans. <Conclusions:< Two new allergens from <L. destructor< have been identified and can now be added to the repertoire of recombinant <L. destructor< allergens. In addition, both these allergens belong to highly conserved protein families and may be important for evaluation of allergenic cross-reactivity. Copyright © 2003 S. Karger AG, Basel