抗CD133/CD3双特异性抗体介导的细胞因子诱导杀伤细胞体外杀伤CD133阳性肿瘤细胞研究  

Effects of cytokine-induced killer cells on CD133 positive cancer cells mediated by anti-CD3/anti-CD133 bispecifi c antibodies

作　　者：吕海燕[1] 刘传杰[1] 黄建华[1] 韩为东[1] 

机构地区：[1]解放军总医院基础医学研究所免疫研究室,北京100853

出　　处：《解放军医学院学报》2015年第12期1227-1231,共5页Academic Journal of Chinese PLA Medical School

摘　　要：目的比较抗CD133/CD3双特异性抗体介导的细胞因子诱导杀伤细胞(cytokine induced killer cells,CIK)与普通CIK对胰腺癌、胃癌和卵巢癌等细胞系的杀伤作用。方法利用杂交瘤技术制备鼠抗人CD133单克隆抗体,用酶联免疫吸附(ELISA)方法检测腹水效价;经过蛋白G亲和层析法纯化得到CD133抗体,在温和的条件下与鼠抗人CD3单抗进行化学偶联,制备抗CD133和抗CD3的双特异性抗体;CCK8试剂盒检测CIK和双特异性抗体介导的CIK杀伤肿瘤细胞系。结果收获腹水效价为1∶50 000,通过标准曲线计算出CD133单抗的浓度为3.2 mg/ml;成功构建了抗CD133/CD3双功能特异性抗体介导的CIK;在相同的效靶比条件下,抗CD133/CD3介导的CIK对表达CD133的胰腺癌细胞系SW1990、胃癌细胞系SGC7901和卵巢癌细胞系A2780有更强的杀伤作用,与单纯CIK相比有显著差异,在效靶浓度为20∶1时,抗CD133/CD3双特异性抗体介导CIK细胞较普通CIK对SW1990细胞株杀伤率提高了30.99%,对SGC7901细胞株杀伤率提高了24.42%(P<0.05)。结论抗CD133/CD3双功能抗体可以显著改善CIK对CD133阳性肿瘤细胞的杀伤作用。Objective To compare the killing effect of anti-CD3/anti-CD133 bispecific antibodies- cytokine-induced killer（Bs AbCIK） with general CIK cells on CD133 positive tumor cells. Methods Hybridoma technique was used to generate human antiCD133 monoclonal antibody, and enzyme-linked immunosorbent assay（ELISA） was utilized to detect the titer of antibody. AntiCD3/anti-CD133 bispecific antibodies were produced through chemical coupling with anti-CD3 and anti-CD133 antibody, which was purified by protein G. The killing effects of Bs Ab-CIK and general CIK on CD133 positive cancer cells were evaluated by CCK-8. Results The titer of anti-CD133 antibody in ascites was 1∶50 000 and the concentration was 3.2 mg/ml. The CIKs bounded by bispecific antibody were established successfully. The experiment results showed that m Bs Ab-CIK had better activity than general CIK in mediating lysis of CD133 positive cells, such as SW1990, 7901 and A2780 cells, in the same condition of effector-to-target ratio. In effector-to-target ratio of 20∶1, the killing effects was enhanced about 30.99% to SW1990 by Bs Ab-CIK comparing to general CIK, and it also enhanced about 24.42% to SGC7901 with significant differences（P〈0.05）. Conclusion The killing effects of CIK to CD133 positive tumor cells can be enhanced significantly by anti-CD3/anti-CD133 bispecific antibodies.

关 键 词：肿瘤干细胞 细胞因子诱导杀伤细胞 双特异性抗体 CD133单抗 CD3单抗 

分 类 号：R730.2[医药卫生—肿瘤]