Immunochemical and biological analysis of allergenicity with excretory-secretory products of Anisakis simplex third stage larva

J. S. Kim, K. H. Kim, S. Cho, H. Y. Park, Sung-Weon Cho, Y. T. Kim, K. H. Joo, J. S. Lee

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Background: Anisakis simplex third stage larvae (L3) are parasites that frequently give rise to allergic responses. The larvae molt into fourth stage larvae (L4), and at each stage they produce L3-excretory-secretory products (L3-ESP) and L4-ESP, respectively, which are different in their main protein constituents. Although the allergenicity of L4-ESP has been investigated by several research groups, research on the allergenicity of L3-ESP has not been carried out by any researcher. In this investigation, the allergenicity and antigenicity of L3-ESP were investigated in comparison with L4-ESP, using rat sera. Methods: Rat sera were produced by L3 oral infection two times with a 9-week interval. Larvae ESP prepared by culture were concentrated and fractioned using lyophilizer and a centrifugal filter device, respectively. Immunochemical analysis was performed using both indirect ELISA and immunoblot. Biological allergenicity was analyzed by RBL-2H3 exocytosis. Results: With the indirect ELISA, the optical density (OD) value of the nonfractioned (NF)-L3ESP was only one third of that of the NF-L4ESP in both specific IgM and IgG. On measuring specific IgE, the OD of NF-L3ESP was less than one tenth of that of NF-L4ESP. In addition, neither antigen nor allergen was shown in NF-L3ESP, but it was shown in NF-L4ESP with immunoblot. However, the biological allergenicity of NF-L3ESP was comparable to that of NF-L4ESP. To demonstrate the presence of any allergen, L3-ESP was fractioned and found to carry twelve visualized allergen bands from 10 to 186 kDa by immunoblot. Conclusions: These results indicate that L3-ESP may include the important allergens necessary to induce the allergy by L3 oral infection, as compared to L4-ESP.

Original languageEnglish
Pages (from-to)320-328
Number of pages9
JournalInternational Archives of Allergy and Immunology
Volume136
Issue number4
DOIs
Publication statusPublished - 2005 Apr 1

Keywords

  • Allergen
  • Anisakis simplex
  • ELISA
  • IgE
  • Mast cells
  • Western blot

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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