Mapping of IgE and IgG 4 antibody-binding epitopes in Cyn d 1, the major allergen of bermuda grass pollen

Han Chih Yuan, Keh Gong Wu, Chun Jen Chen, Song Nan Su, Horng Der Shen, Yann Jang Chen, Ho Jen Peng*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Background: Bermuda grass pollen (BGP) is an important seasonal aeroallergen worldwide which induces allergic disorders such as allergic rhinitis, conjunctivitis and asthma. Cyn d 1 is the major allergen of BGP. This study is aimed to map human IgE and IgG 4 antibody-binding sequential epitopes on Cyn d 1 by dot immunoblotting. Methods: Synthetic peptides (10-mers; 5 overlapping residues) spanning the full length of Cyn d 1 were used for dot immunoblotting to map human IgE and IgG 1-4 antibody-binding regions with sera from BGP-allergic patients. Synthetic peptides with more overlapping residues were used for further mapping. Essential amino acids in each epitope were examined by single amino acid substitution with alanine. Peptides with sequence polymorphism of epitopes of Cyn d 1 were also synthesized to extrapolate their differences in binding capability. Results: Four major IgE-binding epitopes (peptides 15 -1, 21, 33 -2 and 35 +1, corresponding to amino acids 70-79, 101-110, 159-167 and 172-181) and 5 major IgG 4-binding epitopes (peptides 15 -1, 30 -2, 33 -2, 35 +1 and 39, corresponding to amino acids 70-79, 144-153, 159-167, 172-181 and 192-200) were identified. They are all located on the surface of the simulated Cyn d 1 molecule, and three of them are major epitopes for both IgE and IgG 4. Their critical amino acids were all characterized. Major epitopes for human IgG 1 to IgG 4 are almost identical. Conclusions: This is the first study to map the sequential epitopes for human IgE and IgG 4 subclasses in Cyn d 1. It will be helpful for future development in immunotherapy and diagnosis.

Original languageEnglish
Pages (from-to)125-135
Number of pages11
JournalInternational Archives of Allergy and Immunology
Volume157
Issue number2
DOIs
StatePublished - Feb 2012

Keywords

  • Antibody-binding epitope
  • Cyn d 1
  • Dot immunoblotting
  • Synthetic peptide

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