抗人卵巢癌/抗人CD3单链双特异性抗体的生物学活性研究

Biologic Properties of an Anti-human Ovarian Carcinoma/Anti-human CD3 Single Chain Bispecific Antibody

背景与目的目前,常规疗法已难以提高卵巢癌患者的生存率,已有实验数据及临床前试验结果表明,双特异性抗体能有效介导效应细胞对肿瘤细胞的杀伤作用。本研究旨在通过检测抗人卵巢癌/抗人CD3单链双特异性抗体(BHL-I)的生物学活性,为其临床前试验及应用提供实验依据。方法观察BHL-I介导的外周血淋巴细胞(peripheralbloodlymphocyte,PBL)与靶细胞SKOV3结合、MTT法检测外周血单个核细胞(peripheralbloodmononuclearcells,PBMCs)增殖及PBL对靶细胞SKOV3杀伤效应,ELISA法检测杀伤过程中PBL分泌hIFN-γ、hTNF-α变化。结果BHL-I介导的花环形成率(15.7%)显著高于对照组(11.1%)(P<0.01);在抗原存在下,BHL-I显著促进PBMC增殖和PBL对靶细胞的杀伤(P<0.01),且杀伤率与花环形成率呈正相关(r=0.946);杀伤过程中上清液中hIFN-γ、hTNF-α显著增高(P<0.01)。结论BHL-I能介导PBL和SKOV3结合并活化PBL特异杀伤效应,杀伤可能与其hIFN-γ、hTNF-α表达增高有关。

BACKGROUND & OBJECTIVE: Previous routine therapies can' t improve the survival rate of ovarian carcinoma patients. Experimental and pre-clinical data showed that bispecific antibodies could efficiently induce antitumor effect of cytotoxic cells. This study was to investigate the biologic properties of an anti-human ovarian carcinoma/anti-human CD3 single chain bispecific antibody (BHL-I) in vitro, and provide reference for pre-clinical experiment and its application. METHODS: Peripheral blood lymphocytes (PBLs), isolated from peripheral blood of healthy donors, were treated with BHL-I. The conjugation between PBLs and target ovarian carcinoma SKOV3 cells was observed under reverse microscope; the proliferation of peripheral blood mononuclear cells (PBMCs) and cytotoxicity of PBLs to SKOV3 cells were detected by MTT assay; the concentrations of human interferon-γ (hIFN-γ) and human tumor necrosis factor-α (hTNF-α), secreted by PBLs in the process of killing target cells, were detected by ELISA. RESULTS: The rosette (PBL-SKOV3) formation rate was significantly higher in BHL-I group than in control group (15.7% vs. 11.1%, P<0.01). BHL-I significantly enhanced the proliferation of PBLs and cytotoxicity of PBLs to SKOV3 cells in the presence of relative antigen (P<0.01); the cytotoxic rate was positively correlated with the rosette formation rate (r=0.946); the concentrations of hIFN-γ and hTNF-α were significantly increased (P<0.01). CONCLUSION: BHL-I could mediate conjugation between PBLs and SKOV3 cells, and activate the cytotoxicity of PBLs which may relate with up-regulation of hIFN-γ and hTNF-α.