Allergomic study of cypress pollen via combinatorial peptide ligand libraries

J Proteomics. 2012 Dec 21;77:101-10. doi: 10.1016/j.jprot.2012.07.010. Epub 2012 Jul 17.

Abstract

Although Cupressus sempervirens (Cups) pollen represents one of the main aeroallergens in southern Europe, only two Cups allergens have yet been identified and reported: Cup s 1 and Cup s 3. The aim of this study was to identify allergens in cypress pollen using an immuno-proteomic approach. A sequential pollen protein extraction was developed and supplemented by a combinatorial peptide ligand library (CPLL) treatment to select low-abundance species. Control extracts and CPLL eluates have then been resolved by 1-DE and 2-DE gel electrophoresis, blotted and confronted with sera from cypress allergic patients. Extracted proteins including IgE-binding components were identified using nanoLC-MS/MS analysis. A total of 108 unique gene products were identified analyzing the eluates and control loaded onto 1-DE SDS-PAGE. Forty proteins were identified in control samples and 68 supplementary species upon CPLL treatment. Out of the 12 IgE-binding proteins characterized in 2-DE gels, 9 were already reported as allergens in various sources including the two major known allergens of Cupressaceae (groups 1 and 2). Three IgE-binding proteins, not previously reported as allergens, are newly described. The improvement in protein extraction combined with the enrichment of low-abundance species allowed us to extend the repertoire of potential cypress pollen allergens.

Publication types

  • Clinical Trial

MeSH terms

  • Allergens / chemistry*
  • Allergens / immunology*
  • Allergens / isolation & purification
  • Antigens, Plant / chemistry
  • Antigens, Plant / immunology
  • Cupressus / chemistry*
  • Cupressus / immunology
  • Female
  • Humans
  • Hypersensitivity / immunology
  • Immunoglobulin E / chemistry*
  • Immunoglobulin E / immunology
  • Male
  • Peptide Library*
  • Pollen / chemistry*
  • Pollen / immunology
  • Polysaccharide-Lyases / chemistry
  • Polysaccharide-Lyases / immunology

Substances

  • Allergens
  • Antigens, Plant
  • Peptide Library
  • Immunoglobulin E
  • Polysaccharide-Lyases
  • Cup s 1 allergen, Cupressus sempervirens