Chung Young-Bae, Lee Mejeong, Yang Hyun-Jong, Chung Byung-Suk, Lee Shun-Yu, Choi Min-Ho, Hong Sung-Tae
Department of Parasitology, Institute of Endemic Diseases, Seoul National University College of Medicine, Seoul 110-799, Korea.
Korean J Parasitol. 2002 Jun;40(2):83-8. doi: 10.3347/kjp.2002.40.2.83.
The 8 kDa antigenic protein of Clonorchis sinensis was partially purified by ammonium sulfate precipitation and subsequently by a column chromatographic steps. The purified protein was separated into 7 and 8 kDa protein bands through SDS-tricine gel electrophoresis, while the protein was found to migrate to a 8 kDa band in 7.5-15% SDS-PAGE. The molecular weight of the antigen was estimated to be 110 kDa by Superose 6 HR 10/30 gel filtration. The purified antigen strongly reacted with the human sera of clonorchiasis. The hyperimmune sera of BALB/c mice immunized against the 8 kDa protein were reacted with both the crude extract and the excretory-secretory product of adult worms, but not with the metacercarial extract. Immunohistochemical staining demonstrated that the protein was distributed to the tegument and subtegumental cells and also to the seminal receptacle. The present findings suggest that the 8 kDa protein is a partition of the multicomplex protein originating from various organs of adult C. sinensis, and that it is composed of several 7 and 8 kDa proteins.
华支睾吸虫的8 kDa抗原蛋白先用硫酸铵沉淀法进行部分纯化,随后再经柱色谱步骤进一步纯化。通过SDS-三羟甲基氨基甲烷凝胶电泳,纯化后的蛋白被分离成7 kDa和8 kDa的蛋白条带,而在7.5 - 15%的SDS-聚丙烯酰胺凝胶电泳中该蛋白迁移至8 kDa条带。通过Superose 6 HR 10/30凝胶过滤法估计该抗原的分子量为110 kDa。纯化后的抗原与华支睾吸虫病患者血清发生强烈反应。用8 kDa蛋白免疫的BALB/c小鼠的超免疫血清与成虫的粗提物及排泄-分泌产物均发生反应,但与后尾蚴提取物不发生反应。免疫组织化学染色显示,该蛋白分布于虫体皮层和皮层下细胞以及受精囊。目前的研究结果表明,8 kDa蛋白是源自华支睾吸虫成虫各器官的多复合体蛋白的一部分,且由几种7 kDa和8 kDa的蛋白组成。