고려대학교

초록/요약

Background: Anisakis simplex third stage larvae (L3) are the parasites that raise allergy frequently. The larvae molt into fourth stage larvae (L4), and each stage larva produces L3-excretory secretory products (L3ESP) and L4ESP respectively, which are different from each other in main protein constituents. To analyze the allergens to raise the allergy, allergenicity and antigencity of both ESP were investigated using rat sera. Methods: Rat sera were administered by L3-oral infection for two times with nine week interval. Larva ESP prepared by culture in RPMI-1640 media was concentrated and fractioned using lyophilizer and centrifugal filter device respectively. Immunochemical analysis was performed using both by indirect ELISA and by immunoblot. Biological allergenicity was analyzed by RBL-2H3 exocytosis using both administered rat sera and ESP. Results: When specific antibody level for each ESP was measured using indirect ELISA, the level for non-fractioned (NF)-L3ESP was only one third of that for NF-L4ESP both in IgG and in IgM. And, the former was less than one tenth of the latter in IgE. In addition, the former did not show any evident antigen, nor allergen band on immunoblot . However, the biological allergenicity of NF-L3ESP was comparable to that of NF-L4ESP in RBL-2H3 exocytosis. To verify any allergen in L3ESP, L3ESP was fractioned and analyzed immunochemically. In the fractioned L3ESP, twelve allergen bands from 10~186 KDa were evidently visualized on immunoblot. Conclusions: These results indicated that L3ESP may include the important allergens to raise the allergy by L3-oral infection, comparable to L4-ESP.