Molecular cloning, purification, and IgE-binding of a recombinant class I chitinase from Hevea brasiliensis leaves (rHev b 11.0102)

Allergy. 2003 Mar;58(3):246-51. doi: 10.1034/j.1398-9995.2003.00058.x.

Abstract

Background: Class I chitinase in natural rubber latex (NRL) has been assumed to be an important allergen, especially concerning its cross-reactivity with fruits like avocado and banana.

Objectives: The present study aimed to produce a recombinant latex class I chitinase from Hevea brasiliensis leaves and to study its immunoglobulin (Ig)E-binding reactivity.

Methods: A class I chitinase-specific complementary DNA from H. brasiliensis leaves was synthesized, subcloned, sequenced and overexpressed in fusion with the maltose-binding protein (MBP) in Escherichia coli. The IgE-binding reactivity of this protein was studied by the Pharmacia CAP System and by immunoblot experiments using sera from latex-allergic patients.

Results: The rHev b 11.0102 was found to have a length of 295 amino acid residues and contains an N-terminal hevein-like domain with a 56% homology to hevein. Analysis by the CAP method revealed the presence of rHev b 11.0102-specific IgE antibodies in 17 of 58 sera (29%) of IgE-mediated latex-allergic subjects tested. Immunoblot analysis of the MBP-rHev b 11.0102 fusion protein and the MBP carrier protein as a negative control confirmed the IgE-reactivity of rHev b 11.0102.

Conclusion: Due to its IgE-reactivity rHev b 11.0102 represents an allergen of intermediate prevalence in NRL. Its property to cross-react with certain fruits makes it an important supplement in the diagnostic panel of recombinant NRL allergens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Allergens*
  • Amino Acid Sequence
  • Base Sequence
  • Child
  • Chitinases / genetics
  • Chitinases / immunology
  • Chitinases / isolation & purification*
  • Cloning, Molecular
  • Cross Reactions / immunology
  • Female
  • Hevea / chemistry*
  • Hevea / immunology
  • Humans
  • Immunity
  • Immunoglobulin E / immunology*
  • Latex / chemistry*
  • Latex / immunology
  • Latex Hypersensitivity / immunology*
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Plant Proteins / genetics
  • Plant Proteins / immunology
  • Plant Proteins / isolation & purification*
  • Recombinant Proteins

Substances

  • Allergens
  • Latex
  • Plant Proteins
  • Recombinant Proteins
  • Immunoglobulin E
  • Chitinases