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Protein sequence analysis of a novel 103‐kDa Dermatophagoides pteronyssinus mite allergen and prevalence of serum immunoglobulin E reactivity to rDer p 11 in allergic adult patients
Author(s) -
Lee C.S.,
Tsai L.C.,
Chao P.L.,
Lin C.Y.,
Hung M.W.,
Chien A.I.,
Chiang Y.T.,
Han S.H.
Publication year - 2004
Publication title -
clinical and experimental allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.462
H-Index - 154
eISSN - 1365-2222
pISSN - 0954-7894
DOI - 10.1111/j.1365-2222.2004.01878.x
Subject(s) - complementary dna , immunoglobulin e , allergen , house dust mite , mite , immunology , allergy , recombinant dna , biology , microbiology and biotechnology , pyroglyphidae , cdna library , antibody , gene , genetics , botany
Summary Background House dust mites are regarded as important indoor allergens. While the most studies mite allergens are low molecular weight (mw), a high mw Dermatophagoides farinae mite paramyosin (Der f 11) has recently been cloned. We have also cloned a novel high mw Dermatophagoides pteronyssinus (Dp) mite allergen, Der p 11. Objective The aim of this study was to isolate and express a cDNA gene coding for a Der p 11 allergen, to compare the sequence of Der p 11 with other antigens and to evaluate the presence of IgE reactivity to the recombinant protein (rDer p 11) in the sera of allergic adult patients. Methods The full‐length Der p 11 gene was isolated by cDNA library screening, 5′‐3′ rapid amplification of cDNA ends and PCR. The cDNA gene was expressed as a glutathione‐S‐transferase fusion protein in Escherichia coli . The allergenicity of rDer p 11 was tested by human IgE immunodot or immunoblot assay in a large panel of 100 allergic patients with bronchial asthma, allergic rhinitis or eczema. Results Der p 11 is a 2965 bp cDNA gene with a 2625 bp open reading frame coding for a 875 amino acid protein. The deduced amino acid sequence of the Der p 11 showed significant homology with various invertebrate paramyosins. The prevalence of serum IgE reactivity to rDer p 11 on immunodot assay ranged from 41.7% to 66.7% in different allergic patient groups, whereas it was rare in non‐atopic patients with urticaria (18.8%) and in normal individuals (8%). A high frequency (five out of eight) of MAST(Dp) − allergic serum samples had specific IgE‐binding activity to rDer p 11 or its fragments on immunoblot assay, even though their IgE‐binding activity to Dp extract was either weak or negative. Conclusion The 103‐kDa Der p 11 appears to be major Dp mite allergen with a high frequency of IgE reactivity in sera of patients allergic to mites.