摘要
为探究旋毛虫成虫排泄分泌抗原(ES)中丝氨酸蛋白酶对宿主免疫调节作用,用PCR扩增出旋毛虫成虫期特异性丝氨酸蛋白酶基因Zh68,克隆至表达载体pET28a,转化到大肠埃希菌BL21(DE3),经诱导表达后的重组蛋白免疫接种动物获取抗血清。将抗体封闭前后的ES分别作用于S774A.1巨噬细胞,CCK-8检测巨噬细胞的增殖情况,流式细胞仪检测细胞凋亡率。结果发现ES可以抑制S774A.1细胞增殖,并促使其出现凋亡趋势。但用Zh68多抗封闭ES后,这种作用减弱。证实了旋毛虫成虫ES中的丝氨酸蛋白酶可以抑制巨噬细胞增殖,促巨噬细胞出现凋亡趋势,从而为旋毛虫的侵袭与生存提供了有利的环境。
To investigate the effect of serine protease in excretory-secretory antigens（ES） antigens of adult Trichinella spiralis on the host＇s immune system,full length sequence of serine protease gene Zh68 was amplified by PCR.The PCR products was cloned into pET28a vector,and the positive recombinant plasmid was transformed into Escherichia coli BL21（DE3）.The Zh68 antibody was acquired form the rabbit by immunization with recombinant Zh68 protein.ES or ES treated by Zh68 antibody were incubated with S774A.1 macrophages.The number of S774A.1 macrophages was calculated by CCK-8,and the apoptotic rate was determined by flow cytometry（FCM）.The results showed that ES antigens of Trichinella spiralis can inhibit the proliferation of macrophages and promote the apoptosis of S774A.1 macrophages.However,this effect was significantly decreased when ES antigens were treated by Zh68 antibody.We demonsrtated that serine protease in ES antigens of Trichinella spiralis can inhibit the proliferation of macrophages and promote apoptosis of macrophages.
出处
《动物医学进展》
CSCD
北大核心
2011年第10期5-9,共5页
Progress In Veterinary Medicine
基金
国家自然科学基金项目(31030064)
国家杰出青年科学基金项目(30825033)
