用单克隆抗体展示猪链球菌2型溶菌酶释放蛋白在菌体表面的形态和分布  

Configuration and Location of Muramidase Released Protein on Bacterial Cell Surface of Streptococcus suis Type 2 Demonstrated by Monoclonal Antibodies

作　　者：曾巧英[1] 陆承平[1] 

机构地区：[1]南京农业大学农业部动物疫病诊断与免疫重点开放实验室

出　　处：《中国农业科学》2004年第1期143-147,共5页Scientia Agricultura Sinica

基　　金：国家"973"资助项目 (G19990 1190 6)

摘　　要：用猪链球菌 2型江苏分离株HA980 1全菌免疫Balb/c小鼠 ,取其脾细胞和Sp2 / 0细胞融合 ,以电泳纯溶菌酶释放蛋白 (MRP)为抗原 ,间接ELISA筛选MRP抗体阳性孔并用有限稀释法单克隆化 ,剔除与胞壁杂蛋白交叉反应阳性的克隆 ,获得 3株特异针对MRP的单克隆抗体细胞株 3A3、4D5和 6B4。将HA980 1和HEp 2细胞共孵育 ,使细菌粘附于细胞表面 ,用 3株MRP单克隆抗体联合介导FITC标记HA980 1菌体表面的MRP ,激光共聚焦显微观察 ,代表MRP的荧光图像显示 ,MRP是一线形表面大分子 ,一端锚定在细胞壁 ,另一端向空中伸展 ,其分布状态有单在和丛集Three monoclonal antibody hybridomases (MAbs) named 3A3, 4D5 and 6B4 specially directing to muramidase released protein (MRP),a virulence factor of Streptococcus suis type 2(SS2), were produced. In the procedure, Sp2/0 and spleen cells from Balb/c mice immunized with Formalin-killed whole bacteria cells of HA9801,a Jiangsu isolate of SS2, were fused. Then by indirect enzyme-linked immunosorbent assay, the positive hybridomases against MRP were screened using electrophoretically pure MRP as antigen and monoclonalized by limiting dilution, among which, hybridomas clones cross-directing to other cell wall protein potions of HA9801 except MRP were screened out. Bacterial cells of HA9801 were co-incubated with HEp-2 cells to make a adhesion of both cells. Then MRP on cellular surface of HA9801 was labelled with FITC through co-mediation with the three MAbs of MRP. Laser confocal scanning light microscopy revealed that MRP took on a thread-like shape, one end of which was anchored on the cell wall and the other end was free, directing outward singly or in clusters.

关 键 词：单克隆抗体 猪 链球菌2型 溶菌酶 释放蛋白 菌体表面 形态分布 

分 类 号：S852.61[农业科学—基础兽医学]