Immunogenicity of excretory/secretory products from Anisakis simplex worms : cultured under acidic condition
| Autor: | Akihiko, Uchida, Chikaku, Dogasaki, Yasushi, Kawakami |
|---|---|
| Jazyk: | japonština |
| Rok vydání: | 2008 |
| Zdroj: | 麻布大学雑誌 = Journal of Azabu University. :253-257 |
| ISSN: | 1346-5880 |
| Popis: | 一般に寄生虫感染症の各免疫反応に用いられる抗原としては,体成分抗原(SA: Smatic Antigen)が用いられてきたが,近年,虫体の排泄/分泌抗原(ES:Excretory/Secretory Antigen)の有用性が重視され始めている。アニサキス症全症例の約80%の原因寄生虫であるA. simplexを用いて,酸性条件下(pH1)及び中性条件下(pH7)で培養を行い,得られた培養液から排泄/分泌抗原タンパクを抽出し,SDS-PAGEでタンパク質分画を行って,その組成について比較した。また,ラット及びハムスターに生きた虫体を感染させて抗血清を作製し,ELISA及びウエスタンブロット法を行い,両条件下の抗原タンパク成分と反応性に差異が認められるか否かについて検討した。pH1及びpH7の両条件下でA. simplex虫体を培養し,得られたES産物をSDS-PAGEで比較したところ,22kD,25kD,30kDのバンドは,pH1の条件下でのみ検出され,pH7の条件下では検出されなかった。この結果よりES産物のタンパク質組成は,培養液の酸性,中性条件下では抗原タンパク成分に差異が認められ,酸性条件下で虫体が排泄/分泌する抗原は中性条件下とは異なることが明らかとなった。 The somatic antigen of parasitic helminthes has generally been used in the immunodiagnosis of the parasitic infection. In recent years, excretory/secretory(ES) products of worms have been recognized as an important antigen for immunodiagnosis. In our present study, we cultured Anisakis simplex worms in vitro separately under acidic (pH 1) and neutral (pH 7) conditions. The two culture media fluids were then concentrated and used as ES protein antigens. The two antigens were then fractionated and analyzed by SDS-PAGE. Antisera to the worm antigens, produced by orally infecting rats and golden hamsters with live worms, were used to react against the ES antigens in ELISA and Western blotting to examine the immunogenicity of the antigens. It was observed in the Westerb blot that antisera from the infected golden hamstersrecognized different ES protein antigens derived from the acidic culture media when compared to those produced under neutral condition. Thus, we have shown that ES antigens produced by A. simplex worms under acudic condition differ from those produced under neutral condition. |
| Databáze: | OpenAIRE |
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