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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

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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.

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Acknowledgments

We are grateful to Dr. Reiko Homma (Torii Pharmaceutical Co., Ltd., Tokyo, Japan) for kindly giving us monoclonal antibodies. This research was partially supported by the Promotion and Mutual Aid Corporation for Private Schools of Japan, Grant-in-Aid for Matching Fund Subsidy for Private University, and by a Project Grant awarded by the Azabu University Research Services Division.

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Authors and Affiliations

  1. Department of Veterinary Microbiology, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa, 252-5201, Japan

    Kazuki Miyaji, Terumi Yurimoto, Yukari Takase, Hidekatsu Shimakura, Noriaki Okamoto, Akio Kiuchi & Masahiro Sakaguchi

  2. Clinical Research Center for Allergy and Rheumatology, National Sagamihara Hospital, 18-1 Sakuradai, Minami-ku, Sagamihara, Kanagawa, 252-0392, Japan

    Akemi Saito & Hiroshi Yasueda

  3. Department of Molecular Immunology, Institute of DNA Medicine, Jikei University School of Medicine, 3-25-8 Nishi-Shinbashi, Minato-ku, Tokyo, 105-8461, Japan

    Saburo Saito

Authors
  1. Kazuki Miyaji
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  2. Terumi Yurimoto
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  3. Akemi Saito
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  4. Hiroshi Yasueda
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  5. Yukari Takase
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  6. Hidekatsu Shimakura
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  7. Noriaki Okamoto
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  8. Akio Kiuchi
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  9. Saburo Saito
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  10. Masahiro Sakaguchi
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Corresponding author

Correspondence to Masahiro Sakaguchi.

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Supplementary table

Prediction of solvent-accessible regions for all amino acids of mature Cry j 2 protein using NetsurfP (PDF 302 kb)

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Miyaji, K., Yurimoto, T., Saito, A. et al. 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. J Clin Immunol 33, 977–983 (2013). https://doi.org/10.1007/s10875-013-9880-7

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  • DOI: https://doi.org/10.1007/s10875-013-9880-7

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