重组多子小瓜虫抑动抗原ISCOMs制备

Preparation of recombinant immobilization antigen of Ichthyophthirius multifiliis

提取表达于四膜虫（Tetrahymena thermophila）细胞膜表面的多子小瓜虫抑动抗原。通过Western Blotting证实重组多子小瓜虫抑动抗原约为35.7 kD,其免疫原性与多子小瓜虫抑动抗原相似。以Quil-A作为佐剂,采用离心、超声波和透析等方法制备小瓜虫抑动抗原免疫刺激复合物（Iag-ISCOMs）。电镜观察可见Iag-ISCOMs经磷钨酸负染后呈直径30~40 nm的笼格状结构。免疫和攻毒试验证实Iag-ISCOMs对鳗鲡具有一定保护能力,攻毒后存活率达71.4%,对照组存活率为43.9%,结果表明,本实验成功制备了具有一定免疫原性的Iag-ISCOMs。

The recombinant immobilization antigen（Iag） of Ichthyophthirius multifiliis was extracted from mem-brane protein of Tetrahymena thermophila.The molecular weight of the recombinant immobilization antigen is 35.7 kD.In addition,western blotting corroborated that the recombinant Iag had similar immunogenic features to the Iag of I.multifiliis.Iag-ISCOMs were prepared by centrifugation,ultrasonic,dialysis and by using Quil-A as the adjuvant.Results from negative contrast electron microscopy observations indicate that Iag-ISCOM is a cage-like structure with a diameter of 30？40 nm.The Iag-ISCOMs provide protection to the eels based on the tri-als of immunization and challenge.Thus,survival rates of the Iag-ISCOMs and control groups were 71.4% and 43.9%,respectively.Results show that the Iag-ISCOMs with immunogenicity were successfully prepared.