Immunochemical and Biological Analysis of Allergenicity with Excretory-Secretory Products of Anisakis simplex Third Stage Larva

• Published in: International Archives of Allergy and Immunology 2005-04, Vol.136 (4), p.320-328
• Main Authors: Kim, J.S., Kim, K.H., Cho, S., Park, H.Y., Cho, S.W., Kim, Y.T., Joo, K.H., Lee, J.S.
• Format: Article
• Language: English
• Subjects: Allergens - immunology; Allergies; Animals; Anisakis - immunology; Anisakis simplex; Biological and medical sciences; Blotting, Western; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Female; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Immunoglobulin Isotypes; Immunoglobulins; Immunology; Immunopathology; Larva - immunology; Medical sciences; Original Paper; Parasites; Proteins; Rats; Rats, Sprague-Dawley
• ISSN: 1018-2438; 1423-0097; 1365-2567
• DOI: 10.1159/000084225

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.