title: Significance of carbohydrate epitopes in a latex allergen with beta-1,3-glucanase activity.
abstract: BACKGROUND
  One of the latex allergens, Hev b 2, has beta-1,3-glucanase activity. The entire sequence of this allergen is already known. There is one potential N-glycosylation site in this molecule ((27)Asn). Heterogeneous glycosylation of this Asn residue could be a source of the multiplicity of natural Hev b 2. Possible participation of the carbohydrate epitopes of latex beta-1,3-glucanase isoenzymes in their IgE-binding capacity and cross-reactivity was investigated in this study.
  
  
  METHODS
  beta-1,3-Glucanase isoenzymes were separated based on their affinities for concanavalin A. IgE-binding capacity and cross-reactivity were examined by immunoblotting and enzyme-linked immunosorbent assay (ELISA). Sequence heterogeneity among the isoenzymes was probed by peptide mass mapping after lysyl endopeptidase digestion. To clarify the relation to Hev b 2, N-terminal sequencing was performed on a fragmented peptide common to the separated isoenzymes.
  
  
  RESULTS
  Basic beta-1,3-glucanase was subdivided into two glycosylated isoenzymes (GI and GII) and one non-glycosylated isoenzyme (GIII). IgE antibodies in latex-positive sera chiefly recognized the glycosylated isoenzymes. Inhibition ELISA supported the significance of the carbohydrate epitopes for the IgE recognition and cross-reactivity. However, non-glycosylated GIII, as well as GI and GII, produced positive results in a skin prick test. The three beta-1,3-glucanase isoenzymes shared a partial sequence in common with Hev b 2.
  
  
  CONCLUSIONS
  Our results suggest that the carbohydrate epitopes in Hev b 2 homologues are relevant to an in vitro diagnosis of latex allergy and the accompanying cross-reactivity. Carbohydrate epitopes do not necessarily provoke allergic symptoms. Therefore, the actual allergenicity of Hev b 2 and its homologues should be carefully evaluated not only by in vitro IgE tests but also by in vivo tests.
authors.affiliation: Division of Medical Devices, National Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya-ku, Tokyo 158-8501, Japan. yagami@nihs.go.jp
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yagami
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Osuna
authors.forename: Masumi
authors.initials: M
authors.lastname: Kouno
authors.forename: Yuji
authors.initials: Y
authors.lastname: Haishima
authors.forename: Akitada
authors.initials: A
authors.lastname: Nakamura
authors.forename: Zenro
authors.initials: Z
authors.lastname: Ikezawa
mesh_terms: D000328:Adult; D000485:Allergens / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005260:Female; D043326:Glucan 1,3-beta-Glucosidase; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D007527:Isoenzymes / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D020315:Latex Hypersensitivity / Q000209:etiology* / Q000276:immunology; D008297:Male; D008875:Middle Aged; D001617:beta-Glucosidase / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism
chemical_list: D000485:Allergens; D000939:Epitopes; D007527:Isoenzymes; D007073:Immunoglobulin E; D001617:beta-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase