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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 8可在体外介导预激活的外周血淋巴细胞对卵巢上皮癌细胞特异性杀伤
AIM: To investigate the cytotoxicity of bispecific antibody COC183B2 × anti CD28 on human ovarian epithelial carcinoma cell line SKOV3 mediated by preactivated PBMC in vitro. METHODS: COC183B2 × anti CD28 constructed by genetic engineering was induced in E.coli. The activity of the expressed product in combination with ovarian cancer antigen and CD28 antigen was determined by ELISA and incubated with SKOV3 and pre-activated PBMC . After 6h, the cytotoxicity of pre-activated PBMC against SKOV3 cells was determined by non-radioactive cytotoxicity assay and the morphological changes of SKOV3 were observed under microscope. Results: The results of ELISA showed that COC183B2 × anti CD28 could bind with ovarian cancer antigen and CD28 antigen bidirectionally. Cytotoxicity test showed that COC183B2 × anti CD28 could mediate the specific killing of SKOV3 cells by preactivated PBMC in vitro. Under the visible rosette formation, 12h visible tumor cell lysis. CONCLUSION: COC183B2 × anti CD28 can mediate the specific killing of ovarian epithelial cancer cells by preactivated peripheral blood lymphocytes in vitro