title: 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: OBJECTIVE
  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.affiliation: Department of Veterinary Microbiology, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 252-5201, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Miyaji
authors.forename: Terumi
authors.initials: T
authors.lastname: Yurimoto
authors.forename: Akemi
authors.initials: A
authors.lastname: Saito
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yasueda
authors.forename: Yukari
authors.initials: Y
authors.lastname: Takase
authors.forename: Hidekatsu
authors.initials: H
authors.lastname: Shimakura
authors.forename: Noriaki
authors.initials: N
authors.lastname: Okamoto
authors.forename: Akio
authors.initials: A
authors.lastname: Kiuchi
authors.forename: Saburo
authors.initials: S
authors.lastname: Saito
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sakaguchi
mesh_terms: D000485:Allergens / Q000276:immunology*; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D052179:Antigens, Plant / Q000276:immunology; D032485:Cryptomeria / Q000276:immunology*; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology; D010940:Plant Proteins / Q000276:immunology*; D011058:Pollen / Q000276:immunology*
chemical_list: D000485:Allergens; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; C065291:Cry j II protein, Cryptomeria japonica; D018985:Epitopes, B-Lymphocyte; D010455:Peptides; D010940:Plant Proteins; D007073:Immunoglobulin E