日本血吸虫细胞免疫原模拟表位的筛选及初步鉴定  

Screening and identification of mimotopes of Schistosoma japonicum cell-type immunogen

作　　者：李艳琴[1] 蔡力汀[1] 曾庆仁[1] 余平[1] 喻容[1] 

机构地区：[1]中南大学基础医学院细胞与分子生物实验中心

出　　处：《中国病原生物学杂志》2008年第4期276-280,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金资助项目(No.30570952)

摘　　要：目的从噬菌体随机12肽库中筛选日本血吸虫(Schistosoma japonicum,S.j)具有保护性的细胞免疫原模拟表位,为进一步阐明其免疫保护性分子机制提供依据。方法用具有保护性的S.j12d童虫培养细胞(S.jC-12)免疫鼠血清中的IgG对随机12肽库作亲和筛选,随机挑取20个噬菌体克隆用ELISA检测其特异性;对阳性克隆进行测序,采用ELISA和Western blot检测阳性克隆免疫鼠血清中的特异性抗体及其识别天然抗原的特性。结果经过4轮免疫淘洗,洗脱的噬菌体得到了有效富集(16 250倍)。20个噬菌体克隆经检测有15个能被S.jC-12免疫鼠血清识别,有12个序列不同的噬菌体克隆;ELISA检测显示有11个阳性噬菌体克隆诱导小鼠产生IgG应答,其中含精氨酸的阳性噬菌体克隆免疫血清中IgG水平与不含精氨酸克隆免疫鼠比较差异有统计学意义(P<0.01)。结论利用噬菌体展示技术可筛选出S.jC-12免疫原的部分模拟表位且能在小鼠模型中诱导免疫应答。Objective To obtain the specific mimic epitopes of the protective cell-type immunogen of Schistosoma japonicum（S.jC-12） from the peptide display library and provide the bases for illustration of the immunoprotective molecular mechanism. Methods Immunoscreening of a random 12-peptide phage display library was carried out with IgG purified from sera of mice immunized with cultured cells of 12-day S.japonicum.Specificity of 20 clones randomly picked out from the fourth elute was tested by reverse ELISA and the amino acid sequences of positive clones were deduced by sequencing.The specialty of antibodies in sera from mice immunized with positive clones and character of recognition of natural antigens were evaluated by direct ELISA and Western blot,respectively. Results After four rounds of biopanning,the eluted phages were effectively enriched（16 250 times）.15 of 20 clones were recognized by sera from mice immunized with S.jC-12,and,among them,12 clones possess distinct sequences.ELISA result showed that 11 positive clones induced IgG response in immunized mice and significant differences（P〈0.01） were found between sera from mice immunized with the clones containing Arginine residue in their sequences and without Arginine residue. Conclusion Partial specific mimic epitopes of the 12-day cell-type immunogen of S.japonicum could be screened out by utilization of phage display technique and be able to induce immune response in a mouse model.

关 键 词：血吸虫 日本 童虫细胞 噬菌体随机12肽库 表位 

分 类 号：R383.24[医药卫生—医学寄生虫学]