双特异抗体COC183B_2×anti-CD28对卵巢上皮癌细胞杀伤效力的研究

The study of cytotoxicity of bispecific antibody COC183B_2×anti-CD28 againstovarian epithelial carcinoma cells.

目的 :检测双特异抗体 (bispecificantibody ,BsAb)COC183B2 ×anti CD2 8在体外介导预激活的外周血单个核细胞 (PBMC)对卵巢上皮癌细胞系SKOV3的杀伤效力。方法 :在大肠杆菌中对用基因工程方法构建的COC183B2 ×anti CD2 8进行诱导表达 ,用ELISA方法测定表达产物与卵巢癌抗原和CD2 8抗原结合的活性后 ,与SKOV3和预激活的PBMC共孵育。 6h后用非放射性细胞毒分析试剂测定预激活的PBMC对SKOV3细胞毒性作用 ,并在镜下观察SKOV3的形态学改变。结果 :ELISA结果显示 ,COC183B2 ×anti CD2 8可与卵巢癌抗原和CD2 8抗原双向结合 ;细胞毒性试验显示 ,在体外COC183B2 ×anti CD2 8能介导预激活的PBMC特异杀伤SKOV3细胞 ;4h时镜下可见玫瑰花环形成 ,12h可见肿瘤细胞溶解。结论 :COC183B2 ×anti CD2

Objective:To study the specific cytotoxic effects of pre activated human peripheral blood mononuclear cell (PBMC)against ovarian epithelial carcinoma cell line SKOV3 mediated by bispecific antibody COC183B 2×anti CD28.Methods:COC183B 2×anti CD28 constructed by gene engineering was expressed in E.coli.The immunoactivity of COC183B 2×anti CD28 binding withboth CD28 antigen and human ovarian carcinomas antigen OC183B 2 was detectedby ELISA.The cytotoxicity mediated by COC183B 2×anti CD28 was assayed following incubation with SKOV3 and pre activated PBMC at 6 hours,meanwhile,the morphologic changes of SKOV3 were observed under microscopy.Results: The product expressed in E.colicould specifically bind to both anti CD28 antigen and human ovarian carcinomas antigen COC183B 2. In vitro cytotoxicity test showed that COC183B 2×anti CD28 could mediate the specific cytotoxic effects ofpre activated PBMC against SKOV3.The cytomorphology showed that the rosette formed and lysis of SKOV3 occured after 4 hours and 12 hours respectively. Conclusion:COC183B 2×anti CD28 can mediate a specific cytotoxic effect of pre activated PBMC against ovarian carcinoma cell.