INSERM U892, Nantes, France.
MAbs. 2013 Jul-Aug;5(4):587-94. doi: 10.4161/mabs.25077. Epub 2013 May 29.
To take advantage of the large number of well-characterized mouse immunoglobulins (IgGs) for the study of antibody-dependent cell-mediated cytotoxicity (ADCC) in human cells, we armed human cytotoxic lymphocytes with a mouse receptor for the Fc portion of IgG antibodies. The human ΝΚ-92 natural killer cell line was transduced with a mouse receptor gene (mCD16), which was stably expressed on the cell surface (referred to as NK-92 (mCD16) ). When tested against a B-lymphoblastoid cell line (BLCL) coated with mouse anti-CD20 IgG1, IgG2a or IgG2b monoclonal antibodies (mAbs), the newly expressed mouse Fc receptor enabled the NK-92 (mCD16) cells to kill the BLCL by ADCC. Next, using the NK-92 (mCD16) we compared mouse mAbs directed at B lineage specific CD antigens for their ability to induce ADCC against human Epstein-Barr virus- infected B lymphoblastoid (for anti-CD19, -CD20 and -CD21) or against myeloma (for anti-CD38 and -CD138) target cells. Our results demonstrated that the "NK-92 (mCD16) assay" allows convenient and sensitive discrimination of mouse mAbs for their ability to mediate ADCC in a human cellular system. In addition, our results provide examples of dissociation between opsonization and target cell killing through ADCC. These "murinized" human effector cells thus represent a convenient cellular tool for the study of ADCC.
为了利用大量经过充分鉴定的小鼠免疫球蛋白 (IgG) 来研究人细胞中的抗体依赖的细胞介导的细胞毒性 (ADCC),我们用人的细胞毒性淋巴细胞武装了一种针对 IgG 抗体 Fc 部分的小鼠受体。人 NK-92 自然杀伤细胞系被转导了一种小鼠受体基因 (mCD16),该基因稳定地表达在细胞表面(称为 NK-92(mCD16))。当用包被有小鼠抗 CD20 IgG1、IgG2a 或 IgG2b 单克隆抗体 (mAb) 的 B 淋巴细胞样细胞瘤系 (BLCL) 进行测试时,新表达的小鼠 Fc 受体使 NK-92(mCD16)细胞能够通过 ADCC 杀死 BLCL。接下来,我们使用 NK-92(mCD16)比较了针对 B 谱系特异性 CD 抗原的小鼠 mAb 诱导针对人类 EBV 感染的 B 淋巴母细胞 (针对抗-CD19、-CD20 和 -CD21) 或骨髓瘤 (针对抗-CD38 和 -CD138) 靶细胞的 ADCC 的能力。我们的结果表明,“NK-92(mCD16)测定法”允许方便且灵敏地区分小鼠 mAb 介导人细胞系统中的 ADCC 的能力。此外,我们的结果提供了调理作用与 ADCC 介导的靶细胞杀伤之间脱钩的实例。这些“鼠化”的人效应细胞因此代表了研究 ADCC 的便利细胞工具。