抗CD133/CD3双特异性抗体装载的CIK细胞对高表达CD133结直肠癌细胞杀伤效应的探讨

Cytokine-induced killer cells bound with anti-CD3/anti-CD133 bispecific antibodies target CD133-high colorectal cancer cells in vitro

摘要:

目的 探讨抗CD133/CD3双特异性抗体装载的CIK（BsAb-CIK）细胞对高表达CD133结直肠癌细胞的杀伤效应。方法 通过化学偶联制备抗CD133和抗CD3的双特异性抗体。利用CCK8试剂盒检测CIK和BsAb-CIK对高表达CD133结直肠癌细胞系（SW620和HT29）和低表达CD133结直肠癌CIK和BsAb-CIK细胞系（LOVO）的杀伤能力，并检测培养上清中细胞因子分泌水平。构建裸鼠皮下移植瘤模型，随后从腹腔进行CIK和BsAb-CIK细胞输注，1个月后取瘤称重并统计组间肿瘤生长的差异。结果 在不同效靶比条件下（1∶5，1∶10，1∶20），BsAb-CIK细胞对高表达CD133的结直肠癌细胞系SW620和HT29的杀伤作用均明显强于单纯CIK组（P＜0.05），而BsAb-CIK细胞对低表达CD133的结直肠癌细胞系LOVO的杀伤作用与单纯CIK组相比，没有显著性差异（P＞0.05）。BsAb-CIK细胞INF-γ分泌明显高于单纯CIK组（P＜0.05）。与单纯CIK治疗组比较，BsAb-CIK细胞对移植瘤的生长抑制作用更加显著（P＜0.05）。结论 BsAb-CIK细胞能够有效杀伤高表达CD133的结直肠癌细胞。

Abstract:

Objective To determine the killing effects of cytokine-induced killer cells bound with the bispecific antibodies for CD3 and CD133(BsAb-CIK) on CD133-high expression colorectal cancer cells. Methods The bispecific antibodies were generated by chemical coupling with anti-CD3 and anti-CD133 antibodies. CCK-8 kit was used for comparing the killing effects of CIK and BsAb-CIK on CD133-high expression colorectal cancer cell lines (SW620 and HT29) and CD133-low expression colorectal cancer cell line (LOVO). Cytokines secreted in the supernatant were detected. The xenograft for nude mice was established, and then given CIK or BsAb-CIK cells infusion via abdominal cavity. Tumor weight were compared between the groups after 1 month. Results At the different ratio of effector cells to target cells (1∶5, 1∶10, 1∶20), the cytotoxic effects of BsAb-CIK cells on CD133-high expression colorectal cancer cells (SW620 and HT29) were stronger than that of general CIK cells (P＜0.05), but there was no significant difference on CD133-low expression colorectal cancer cells (LOVO). The secretary amount of IFN-γ was higher in the supernatant of BsAb-CIK cells than the general CIK cells when killing experiments were performed. The treatment with BsAb-CIK cells resulted in a more remarkable inhibition on xenograft tumor growth than that with general CIK cells (P＜0.05). Conclusion The BsAb-CIK cells can effectively kill CD133-high expression colorectal cancer cells.