Alvarez-Rueda Nidia, Leprieur Stéphanie, Clémenceau Béatrice, Supiot Stéphane, Sébille-Rivain Véronique, Faivre-Chauvet Alain, Davodeau François, Paris François, Barbet Jacques, Aubry Jacques, Birklé Stéphane
Inserm, Université de Nantes, Nantes Atlantique Universités U601, Département de Recherche en Cancérologie, 9 quai Moncousu, F-44093 Nantes, France.
Clin Cancer Res. 2007 Sep 15;13(18 Pt 2):5613s-5620s. doi: 10.1158/1078-0432.CCR-07-1057.
We previously generated a mouse monoclonal antibody (mAb) specific for the tumor-associated GD2 ganglioside antigen. Here, we describe the development of a chimeric anti-GD2 mAb for more effective tumor immunotherapy.
We cloned the cDNA encoding the immunoglobulin light and heavy chains of the 60C3 anti-GD2 mAb, and constructed chimeric genes by linking the cDNA fragments of the variable regions of the murine light and heavy chains to cDNA fragments of the human kappa and gamma1 constant regions, respectively.
The resultant chimeric anti-GD2 mAb, c.60C3, showed identical binding affinity and specificity to that of its murine counterpart. Both c.60C3 and 60C3 were rapidly internalized by tumor cells at 37 degrees C. When human serum and human natural killer cells were used as effectors in complement-mediated cytotoxicity and antibody-dependent cell cytotoxicity, respectively, c.60C3 was more effective in killing GD2-expressing tumor cells. However, c.60C3 was ineffective at inducing cell death by apoptosis, although binding of 60C3 induced apoptotic death in vitro. In an in vivo, GD2-expressing, syngeneic tumor model, i.v. injection of c.60C3, but not of 60C3, significantly suppressed tumor growth in mice (P<0.0005).
Immune effector functions mediated by this antibody and its potentially reduced immunogenicity make chimeric c.60C3 a promising therapeutic agent against neuroectodermic tumors.
我们之前制备了一种针对肿瘤相关GD2神经节苷脂抗原的小鼠单克隆抗体（mAb）。在此，我们描述一种用于更有效肿瘤免疫治疗的嵌合抗GD2单克隆抗体的研发情况。
我们克隆了编码60C3抗GD2单克隆抗体免疫球蛋白轻链和重链的cDNA，并通过将鼠轻链和重链可变区的cDNA片段分别与人κ和γ1恒定区的cDNA片段连接，构建了嵌合基因。
所得的嵌合抗GD2单克隆抗体c.60C3与其鼠源对应物表现出相同的结合亲和力和特异性。在37℃时，c.60C3和60C3均能被肿瘤细胞迅速内化。当分别用人血清和人自然杀伤细胞作为补体介导的细胞毒性和抗体依赖性细胞毒性的效应细胞时，c.60C3在杀伤表达GD2的肿瘤细胞方面更有效。然而，尽管60C3的结合在体外可诱导凋亡性细胞死亡，但c.60C3在诱导细胞凋亡性死亡方面无效。在一个体内表达GD2的同基因肿瘤模型中，静脉注射c.60C3而非60C3能显著抑制小鼠肿瘤生长（P<0.0005）。
该抗体介导的免疫效应功能及其潜在降低的免疫原性使嵌合c.60C3成为一种有前景的抗神经外胚层肿瘤治疗药物。
