Purification and characterization of Lep d I, a major allergen from the mite Lepidoglyphus destructor

Abstract

A major allergen of the storage mite Lepidoglyphus destructor (Lep d I) has been purified by affinity chromatography using an anti-Lep d I monoclonal antibody. The purity of the protein obtained by this procedure was assessed by reverse-phase HPLC. Lep d I displayed a molecular weight of 14 kD on SDS-PAGE under non-reducing conditions, and 16 kD in the presence of a reducing agent. Analytical IEF revealed a little charge microheterogeneity, showing three bands with pIs 7.6-7.8. Purified Lep d I retained IgE-binding ability, as proved by immunoblotting experiments after SDS-PAGE and RAST with individual sera from L. destructor-sensitive patients. Results from the latter technique demonstrated that 87% of L. destructor-allergic patients had specific IgE to Lep d I, and a good correlation between IgE reactivity with L. destructor extract and Lep d I was found. In addition, RAST inhibition experiments showed that IgE-binding sites on Lep d I are major L. destructor-allergenic determinants, since Lep d I could inhibit up to 75% the binding of specific IgE to L. destructor extract; on the other hand, Lep d I did not cross-react with D. pteronyssinus allergens.

Distribution of dust‐mite allergens (Lep d 2, Der p 1, Der f 1, Der 2) in pig‐farming environments and sensitization of the respective farmers

“…However, van Hage-Hamsten et al (5) suggested that each of the storage mites and D. pteronyssinus possess unique allergens. Ventas et al (11) also found that the major allergen of Lep d 2 is not responsible for allergenic cross-reactivity between L. destructor and D. pteronyssinus. One explanation is that most individuals were exposed to both storage mites and house-dust mites and became sensitized to both.…”

“…The main L. destructor allergen, Lep d 2, has been puri®ed and characterized (11,12), and it has become possible to measure Lep d 2, as well as group 1 and group 2 allergens of Dermatophagoides (13±17), by ELISA.…”

Allergen cross‐reactivity between house‐dust mites and other invertebrates

“…Der 1 is inhibited to the same extent by D. pteronyssinus and L. destructor extracts in inhibition tests, while the cross-reactivity between Der 2 and L. destructor was found to be less extensive (77). Other research did not ®nd any Lep d 1 inhibition of HDM-speci®c IgEbinding (80). The amino-acid sequence homology of Der 2 and Lep d 2 is 43%.…”

Cross-reactivity between storage and dust mites and between mites and shrimp

“…However, some studies have shown some cross-inhibition of IgE binding between storage mites and house dust mite extracts (Chew et al 1999;Harfast et al 1992;Johansson et al 1991Johansson et al , 1994Luczynska et al 1990;Puerta et al 1993;van der Heide et al 1998). Still others have found little cross-allergenicity between storage mites and between storage mite extracts and house dust mite extracts or allergens (Chew et al 1999;Johansson et al 1991;Puerta et al 1993;van Hage-Hamsten et al 1987;Ventas et al 1992).…”