Analysis of conformational and sequential IgE epitopes on the major allergen Cry j 2 of Japanese cedar (Cryptomeria japonica) pollen in humans by using monoclonal antibodies for Cry j 2.

Abstract

PURPOSE:

Japanese cedar (Cryptomeria japonica; CJ) pollinosis is a type I allergy induced by CJ pollen, and Cry j 2 is one of the major allergens in this pollen. In a previous study, we analyzed IgE epitopes on Cry j 2 in humans by using synthetic peptides. The main purpose of this study was to identify B-cell epitopes on Cry j 2 in patients with CJ pollinosis by using monoclonal antibodies (mAbs) for Cry j 2.

METHODS:

We used ELISA with mAbs for the epitope analysis. Sera samples were collected from 80 patients with CJ pollinosis, and allergenic epitopes for mAbs and human IgE were identified using ELISA with synthetic peptides. The importance of the epitopes for human IgE was analyzed using an inhibition ELISA.

RESULTS:

Four independent epitopes (epitope #1, #2, #3, and #4) were identified on Cry j 2 with the use of mAbs. Epitope #3 and #4, corresponding to peptides No. 25 and No. 33, respectively, were newly determined as epitopes for mAbs and human IgE. Inhibition ELISA showed that not only epitope #2 (sequential) but epitope #1 (conformational) may play an important role in the CJ pollinosis.

CONCLUSIONS:

Our results revealed 4 epitopes, including two new ones, on Cry j 2. We also found that inhibition ELISA with appropriate mAbs could be a viable method of evaluating the importance of the conformational and sequential epitopes for human IgE. These results are beneficial for the development of safer and more efficient therapeutic strategies for treating CJ pollinosis.

• Authors: Kazuki Miyaji, Terumi Yurimoto, Akemi Saito, Hiroshi Yasueda, Yukari Takase, Hidekatsu Shimakura, Noriaki Okamoto, Akio Kiuchi, Saburo Saito, Masahiro Sakaguchi
• Journal: Journal of clinical immunology (J Clin Immunol) Vol. 33 Issue 5 Pg. 977-83 (Jul 2013) ISSN: 1573-2592 [Electronic] Netherlands
• PMID: 23504147 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Allergens
• Antibodies, Monoclonal
• Antigens, Plant
• Cry j II protein, Cryptomeria japonica
• Epitopes, B-Lymphocyte
• Peptides
• Plant Proteins
• Immunoglobulin E

Topics

• Allergens (immunology)
• Amino Acid Sequence
• Antibodies, Monoclonal (immunology)
• Antigens, Plant (immunology)
• Cryptomeria (immunology)
• Epitopes, B-Lymphocyte (immunology)
• Humans
• Immunoglobulin E (immunology)
• Molecular Sequence Data
• Peptides (immunology)
• Plant Proteins (immunology)
• Pollen (immunology)