日本血吸虫保护性单克隆抗体SSj14识别模拟表位的筛选及其免疫原性分析  

Screening and immunogenicity analysis of mimotopes of protective monoclonal antibody SSj14 against Schistosoma japonicum

作　　者：傅志强[1] 蔡学忠[1] 刘金明[1] 石耀军[1] 李浩[1] 吴祥甫[2] 蔡幼民[1] 林矫矫[1] 

机构地区：[1]中国农业科学院上海兽医研究所农业部动物寄生虫学重点开放实验室,上海200232 [2]中国科学院上海生化与细胞生物研究所,上海200032

出　　处：《中国兽医科学》2007年第8期645-649,共5页Chinese Veterinary Science

基　　金：国家青年自然科学基金项目(30100138);上海市启明星计划项目(04QMX1462)

摘　　要：为分析日本血吸虫保护性单抗SSj14的模拟表位,观察其免疫保护作用,利用SSj14单抗筛选富集噬菌体的随机六肽库,应用免疫印迹法确定阳性克隆并分析其核苷酸序列,采用竞争ELISA法分析阳性克隆的抗原性,用筛选的阳性噬菌体克隆免疫小鼠进行日本血吸虫病免疫保护试验,计算减虫效果,并用ELISA法检测小鼠免疫前后的特异性抗体水平。结果显示,经3轮富集筛选后,免疫印迹法鉴定到4个阳性克隆,它们对SSj14单抗的抑制率分别为20.0%、40.8%、58.6%、61.5%,并分别在小鼠中诱导了7.42%、9.43%、11.74%、22.30%的减虫率。用噬菌体克隆免疫的小鼠均产生了较高水平的特异性抗体。结果表明,筛选到的阳性噬菌体克隆和SSj14单抗有较高的亲和力,并诱导了抗小鼠日本血吸虫的部分保护效果。To analyze peptides mimicking epitope of a protective monoclonal antibodies（McAb） SSjl4 against Schistosoma japonicum and investigate their immune-protective effects,a phage random 6-peptide library was screened using the purified McAb SSjl4, and some clones were picked up. Western-blotting analysis confirmed that 4 positive clones were obtained. The epitope of each positive clone was detected by sequence analysis. The suppression rates of the 4 positive clones to McAb SSjl4 were 20.0%, 40. 8%, 58. 6% and 61.5% in the competitive ELISA,respectively. They induced adult worm reduction by 7. 420/4o, 9. 430/oo ,11.74% and 22.30o/oo in mice,respectively. Meanwhile,levels of the specific antibody in sera of the vaccinated mice was higher than that in the blank control. These results showed that the 4 positive phage clones had high affinity with McAB SSjl4,and provided partial protection against schistosomiasis.

关 键 词：日本血吸虫 噬菌体展示 SSj14单抗 模拟表位 

分 类 号：S852.735[农业科学—基础兽医学] Q813.2[农业科学—兽医学]