日本血吸虫成虫和虫卵可溶性抗原及其组分抗原的研究

目的:探讨日本血吸虫雄虫、雌虫、虫卵的可溶性抗原雄虫(AWA-m、雌虫AWA-f、虫卵SEA)及其组分抗原的特性.方法:用DE22纤维素层析柱对日本血吸虫AWA-m,AWA-f,SEA抗原进行纯化,得到相应组分抗原.分别以SDS-PAGE和Western blot对组分抗原进行全面分析,并与相应未经纯化的可溶性抗原作比较.结果:经DE22纤维素层析处理的AWA-m,AWA-f,SEA,均能获得含多种蛋白的组分抗原.SDS-PAGE结果表明,AWA-m见10条主带和9条次带,其中Mr 37 000,28 000,25 000为特异性条带,出现8条免疫反应阳性带;而AWA-m组分抗原为9条蛋白带和6条免疫反应阳性带.AWA-f见15条蛋白带,其中Mr 26 500为特异性条带,组分抗原为8条蛋白带.AWA-f粗抗原及其组分抗原均见9条反应带.SEA见8条主带和10条次带,13条为免疫反应阳性条带;SEA组分抗原见11条带,其中9条为免疫反应阳性带.结论:AWA-m,AWA-f,SEA经DE22纤维素层析纯化获得组分抗原,方法简单、可靠.该组分抗原含有粗抗原的主要免疫反应成分,去除了多种与日本血吸虫病免疫反应无关的蛋白.

Analysis on soluble antigen and its fraction antigen of Schistosoma japonicum adult and egg

Objective: To explore the characteristics of soluble male adult worm antigens(AWA-m),female adult worm antigens(AWA-f),egg antigens(SEA)and their fraction antigens of Schistosomia japonicum. Methods: The fraction antigens were isolated from AWA-m,AWA-f and SEA on DE22 cellulose column,analyzed by means of SDS-PAGE and western blot(WB).Results: The three types of fraction antigens could be obtained from AWA-m,AWA-f and SEA on DE22 cellulose column.AWA-m and its fraction antigens were separated into 19 and 9 distinct bands in SDS-PAGE gel and revealed 8 and 6 immune bands on western blot respectively.AWA-f and its fraction antigens were separated into 15 and 8 distinct bands in SDS-PAGE gel and displayed 9 immune bands on WB.Most of AWA-m and AWA-f protein bands were similar,but Mr 37 000 and Mr 28 000 and Mr 25 000 bands were only appeared in AWA and Mr 26 500 band was seen in AWA-f.SEA and its fraction antigens were separated into 18 and 11 distinct bands respectively in SDS-PAGE gel and showed 13 and 9 immune reaction bands on WB. Conclusion: The results suggest that our method for obtaining fraction antigens is simple and reliable by purifying AWA-m,AWA-f and SEA with DE22 cellulose.The fraction antigens contained the main immune components of crude soluble antigens and wiped out the major proteins irrespective with immune reaction of Schistosoma japonicum.