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

Clin Exp Allergy. 2004 Mar;34(3):354-62. doi: 10.1111/j.1365-2222.2004.01878.x.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Allergens / genetics*
  • Allergens / immunology
  • Amino Acid Sequence
  • Antigens, Dermatophagoides / genetics
  • Antigens, Dermatophagoides / immunology
  • Arthropod Proteins
  • Asthma / immunology
  • Base Sequence
  • Case-Control Studies
  • Dermatophagoides pteronyssinus / immunology*
  • Eczema / immunology
  • Female
  • Humans
  • Hypersensitivity / immunology*
  • Immunoblotting
  • Immunoglobulin E / blood*
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Recombinant Proteins / immunology
  • Rhinitis, Allergic, Perennial / immunology
  • Sequence Analysis, Protein*
  • Urticaria / immunology

Substances

  • Allergens
  • Antigens, Dermatophagoides
  • Arthropod Proteins
  • Dermatophagoides farinae antigen f 2
  • Dermatophagoides pteronyssinus antigen p 2
  • Recombinant Proteins
  • Immunoglobulin E