Molecular cloning, expression and immunological characterisation of Pas n 1, the major allergen of Bahia grass Paspalum notatum pollen

Mol Immunol. 2008 Dec;46(2):286-93. doi: 10.1016/j.molimm.2008.08.267. Epub 2008 Sep 24.

Abstract

Bahia grass, Paspalum notatum, is a clinically important subtropical grass with a prolonged pollination season from spring to autumn. We aimed to clone and characterise the major Bahia grass pollen allergen, Pas n 1. Grass pollen-allergic patients presenting to a tertiary hospital allergy clinic were tested for IgE reactivity with Bahia grass pollen extract by skin prick testing, ImmunoCAP, ELISA and immunoblotting. Using primers deduced from the N-terminal peptide sequence of a group 1 allergen of Bahia grass pollen extract separated by two-dimensional gel electrophoresis, the complete Pas n 1 cDNA was obtained by rapid amplification of cDNA ends and cloned. Biological relevance of recombinant Pas n 1 expressed in Escherichia coli was assessed by serum IgE reactivity and basophil activation. Twenty-nine of 34 (85%) consecutive patients presenting with grass pollen allergy were skin prick test positive to Bahia grass pollen. The Pas n 1 cDNA has sequence homology with the beta-expansin 1 glycoprotein family and is more closely related to the maize pollen group 1 allergen (85% identity) than to ryegrass Lol p 1 or Timothy grass Phl p 1 (64 and 66% identity, respectively). rPas n 1 reacted with serum IgE in 47 of 55 (85%) Bahia grass pollen-allergic patients, activated basophils and inhibited serum IgE reactivity with the 29 kDa band of Bahia grass pollen extract. In conclusion the cDNA for the major group 1 allergen of the subtropical Bahia grass pollen, Pas n 1, was identified and cloned. rPas n 1 is immunologically active and is a valuable reagent for diagnosis and specific immunotherapy of grass pollen allergy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens / genetics*
  • Allergens / immunology*
  • Asthma / immunology
  • Base Sequence
  • Basophils / immunology
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • DNA, Complementary / immunology
  • Female
  • Humans
  • Immunoglobulin E / immunology
  • Male
  • Molecular Sequence Data
  • Paspalum / genetics*
  • Paspalum / immunology*
  • Plant Proteins / genetics*
  • Plant Proteins / immunology*
  • Pollen / genetics*
  • Pollen / immunology*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Rhinitis, Allergic, Seasonal / immunology
  • Skin Tests

Substances

  • Allergens
  • DNA, Complementary
  • Plant Proteins
  • Recombinant Proteins
  • Immunoglobulin E