Immunologic characterization and allergenicity of recombinant Tyr p 3 allergen from the storage mite Tyrophagus putrescentiae

Int Arch Allergy Immunol. 2009;150(1):15-24. doi: 10.1159/000210376. Epub 2009 Apr 2.

Abstract

Background: The involvement of the storage mite Tyrophagus putrescentiae in allergies has been increasingly reported in many countries. Molecular analysis has shown that group 3 mite allergens are homologous to trypsin. Similar allergens have not been identified in T. putrescentiae. Our aims were to characterize group 3 allergens in T. putrescentiae and to investigate their significance in allergenicity.

Methods: cDNAs of PreTyr p 3 and rTyr p 3 from T. putrescentiae were cloned and expressed in Escherichia coli. Native Tyr p 3 (nTyr p 3) was purified from spent growth medium with an affinity column coupling of antibody. Biological activities of rTyr p 3 were compared with nTyr p 3 in terms of IgE activity, enzymatic activity and histamine release.

Results: Full-length cDNA of PreTyr p 3 encodes a 285-amino acid trypsin-like protease and acquires enzymatic activity after removing the pre- and pro-sequences. rTyr p 3 is a 26-kDa protein with equivalent IgE reactivity but weaker enzymatic activity than that of nTyr p 3. A limited level of cross-reactivity has been found between rTyr p 3, Der p 3 and Blomia. Eight of 10 T. putrescentiae-sensitized individuals showed >50% histamine release after triggering with rTyr p 3.

Conclusions: Our studies demonstrate that Tyr p 3 is a frequent allergen (58%) in T. putrescentiae-sensitized patients. Since rTyr p 3 displays equivalent biological activities as nTyr p 3, the role of group 3 allergens can be studied using rTyr p 3 to elucidate the pathogenic effects and diagnostic applications of Tyr p 3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acaridae / genetics*
  • Acaridae / immunology*
  • Allergens / genetics
  • Allergens / immunology*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western
  • Enzyme-Linked Immunosorbent Assay
  • Genes, Insect*
  • Humans
  • Hypersensitivity / genetics
  • Hypersensitivity / immunology
  • Immunoglobulin E
  • Insect Proteins / genetics*
  • Insect Proteins / immunology*
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology

Substances

  • Allergens
  • Insect Proteins
  • Recombinant Proteins
  • Immunoglobulin E