杆状病毒表达的hZP3 B细胞表位嵌合肽的免疫活性研究  

Studies on the Immunocytochemistry of hZP3 B-cell Epitope Chimeric Peptide Exp ressed by Baculovirus Expression System

作　　者：曹佐武[1] 杨林[2] 郑辉[3] 刘穗玲[4] 肖銮娟[1] 谢琪璇[1] 王自能[5] 

机构地区：[1]暨南大学生殖免疫研究中心,广东广州510632 [2]中山大学生物医药中心,广东广州510275 [3]暨南大学医学院生理教研室,广东广州510632 [4]中山大学第三附属医院,广东广州510630 [5]暨南大学医学院妇产科,广东广州510632

出　　处：《中山大学学报（自然科学版）》2003年第5期70-73,共4页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：国家自然科学基金资助项目(30271370);广东省自然科学基金资助项目(010398)

摘　　要：为研究杆状病毒系统表达的人透明带ZP3的B细胞表位嵌合肽(BCP)的免疫原性及其抗血清与人透明带的免疫交叉反应,重组z3bcp病毒用于感染sf9细胞以表达BCP。Tricine＿SDS＿PAGE用于分离感染液中的小相对分子质量表达产物,表达产物用于免疫小鼠制备抗血清,免疫细胞化学试验用于检验抗血清与人卵泡透明带的交叉反应性。结果显示,sf9细胞被z3bcp病毒感染后,培养液中分离出一条相对分子质量约为6000的特异带,而正常sf9细胞中没有等同的带。免疫细胞化学试验结果显示,该特异肽的抗血清能与人透明带特异结合,表明人透明带BCP嵌合肽在杆状病毒系统中成功表达,表达产物具有抗原性,抗血清能与人透明带特异结合。To produce the recombinant B_cell epitope chimeric peptide (BCP) of human zona pellucida 3(hZP3) and study its immunogenicity and cross reactivity with human zona pellucida,recombinant virus (z3bcp virus) containing hZP3 BCP DNA fragment was used to infect sf9 cells to express BCP. Tricine_SDS_PAGE was performed to resolve the specific product of small molecular mass. The antiserum against recombinant BCP was produced by immunization of mice. Immunocytochemical assay was performed to test the cross_reactivity of BCP with human zona pellucida. After sf9 cells were infected by the recombinant virus, a specific peptide band was separated from the medium of the z3bcp virus_infected cells by PAGE. The molecular weight of the specific peptide is about 6 000, as compared to the standard protein. No identical band was observed in the sample of the normal sf9 cells. Immunocytochemical assay showed that antibody against the specific peptide is specifically bound to the zona pellucida on humans ovary sections. These results suggest that the hZP3 BCP gene was successfully expressed in the sf9 cells. The recombinant BCP is antigenic in animals to induce antibody cross reacted with human zona pellucida.

关 键 词：人透明带3 嵌合肽 免疫细胞化学试验 杆状病毒 透明带避孕疫苗 

分 类 号：R392-33[医药卫生—免疫学] R169.41[医药卫生—基础医学]