Lep d 2 polymorphisms in wild and cultured Lepidoglyphus destructor mites

• Published in: European journal of biochemistry Vol. 270; no. 4; pp. 646 - 653
• Main Authors: Kaiser, Liselotte, Gafvelin, Guro, Johansson, Eva, van Hage‐Hamsten, Marianne, Rasool, Omid
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01.02.2003
• Subjects: allergen; Allergens - genetics; Allergens - pharmacology; Amino Acid Sequence; Animals; Blotting, Southern; Cloning, Molecular; dust mite; Enzyme-Linked Immunosorbent Assay; Immunoblotting; Immunoglobulin E - metabolism; Interferon-gamma - metabolism; Interleukin-5 - metabolism; Lepidoglyphus destructor; Lep d 2; Medicin och hälsovetenskap; Mites - genetics; Molecular Sequence Data; Mutagenesis, Site-Directed; Polymerase Chain Reaction; polymorphism; Polymorphism, Genetic - genetics; Proteins - genetics; Proteins - pharmacology; Sequence Homology, Amino Acid; T-Lymphocytes - drug effects; T-Lymphocytes - immunology
• ISSN: 0014-2956; 1432-1033
• DOI: 10.1046/j.1432-1033.2003.03412.x

We have previously cloned, expressed and characterized two variants of the major allergen Lep d 2 from cultured Lepidoglyphus destructor mites. These variants, Lep d 2.0101 and Lep d 2.0201, differ at 13 amino acid positions. In this study we investigated Lep d 2 sequence diversity between wild and cultured mites. PCR, Southern blot and DNA sequence analysis revealed the presence of two different Lep d 2 genes, one with and one without an intron. In addition, two new variants of Lep d 2, Lep d 2.0102 and Lep d 2.0202, were found at different frequencies in wild and cultured mites. When we expressed the Lep d 2 variants and compared their IgE binding properties by ELISA inhibition, we found that Lep d 2.0102 was a more potent inhibitor than Lep d 2.0101, and to a lesser extent Lep d 2.0202 was more potent than Lep d 2.0201. Long‐term cultures of peripheral blood mononuclear cells were used to assess the ability of the expressed Lep d 2 variants to induce cytokine release. Although cells from different individuals released different amounts of interferon‐γ and interleukin‐5, no consistent cytokine release pattern could be linked to any specific Lep d 2 variant. In conclusion, we show that both cultured and wild Lepidoglyphus destructor mites contain the same pattern of polymorphism. Furthermore, this Lep d 2 sequence diversity seems not to have any significant impact on the allergens IgE binding or its ability to induce T cell cytokine release.