Wu T_(11)(CD2)-小鼠抗人绵羊红细胞受体杂交瘤细胞系的建立

本文报道用T细胞系JMN1免疫BALB/c小鼠,然后取免疫脾细胞与小鼠骨髓瘤细胞系X 63Ag8.653融合。用扁桃腺冰冻切片免疫酶染色法,筛选出一株稳定分泌IgG1亚类的抗绵羊红细胞受体(ER)单扰(McAb)的杂交瘤—WuT11。通过E玫瑰花结形成抑制试验,膜抗原抽提物对WuT11的间接免疫荧光阻断试验,对外周血单个核细胞(PBMC)及其E~ 、E~-层细胞,T、B和非T非B细胞系的反应特征以及FACS的反应曲线等,证明WuT11与OKT11和anti-CCT3相类似,竞争抑制试验证明WuT11与anti-CCT3识别同一表位。用WuT11进行亲和层析,提取的相应抗原经SDS-PAGE,测得分子量为50KDa的糖蛋白,证明WuT11属CD_2,为识别人ER的单抗。

WuTll-Establishment of Mouse Hybridoma Secreted Monoclonal Antibody against Sheep Erythrocyte Receptor on Human T Cells

The preparation of mouse monoclonal antibody,WuT 11,against sheep erythrocyte receptor on human T cells by fusing spleen cells of BALB/c mouse immunized with T cell line JMN1 with X63Ag 8.653 was reported. It could block E rossette formation of huamn T cells with sheep erythrocytes.WuTll reacted exclusively with cells of T cell lineage including thymocytes, peripheral blood T cells and T cell lines(JMN1,CEM,HSB2)and did not react with peripheral blood B cells,Granulocytes, B cell lines (H2LCL,EB-3, 1788, Raji, Daudi, ERL) non-T , non-B cell lines(Reh, Km-3). K562 and platelets. All of WuTll, anti-CCTs and OKT11 gave similar figure of FACS and staining of freeze section of tonsil and thymus. Anti-CCT3 could compete with FITC-WuTll for same epitope on human T cell surface. WuTll recognizied approximately 50KDa molecular on PBMC. The important role and potential application of WuTll were discussed.