Biomonitor SAS, Villejuif Bio Park, 1 Mail du Professeur Georges Mathé, 94800 Villejuif, France.
J Immunol Res. 2017;2017:3908289. doi: 10.1155/2017/3908289. Epub 2017 Sep 27.
Novel ADCC effector cells expressing the V-variant or F-variant of FcRIIIa (CD16a) and firefly luciferase under the control of a chimeric promoter incorporating recognition sequences for the principal transcription factors involved in FcRIIIa signal transduction, together with novel target cells overexpressing a constant high level of the specific antigen recognized by rituximab, trastuzumab, cetuximab, infliximab, adalimumab, or etanercept, confer improved sensitivity, specificity, and dynamic range in an ADCC assay relative to effector cells expressing a NFAT-regulated reporter gene and wild-type target cells. The effector cells also contain a normalization gene rendering ADCC assays independent of cell number or serum matrix effects. The novel effector and target cells in a frozen thaw-and-use format exhibit low vial-to-vial and lot-to-lot variation in their performance characteristics reflected by CVs of 10% or less. Homologous control target cells in which the specific target gene has been invalidated by genome editing providing an ideal control and a means of correcting for nonspecific effects were observed with certain samples of human serum. The novel effector cells and target cells expressing noncleavable membrane-bound TNF have been used to quantify ADCC activity in serum from patients with Crohn's disease treated with infliximab and to relate ADCC activity to drug levels.
新型 ADCC 效应细胞表达 FcRIIIa（CD16a）的 V 变体或 F 变体，以及萤火虫荧光素酶，受包含涉及 FcRIIIa 信号转导的主要转录因子识别序列的嵌合启动子的控制，以及新型靶细胞过度表达特异性抗原的恒定高水平，该抗原被利妥昔单抗、曲妥珠单抗、西妥昔单抗、英夫利昔单抗、阿达木单抗或依那西普识别，与表达 NFAT 调节报告基因和野生型靶细胞的效应细胞相比，在 ADCC 测定中提高了灵敏度、特异性和动态范围。效应细胞还包含一个归一化基因，使 ADCC 测定不受细胞数量或血清基质效应的影响。新型效应细胞和靶细胞以冷冻解冻即用格式存在，其性能特征的瓶间和批间变化很小，CV 值为 10%或更低。在某些人血清样本中观察到经过基因组编辑使特定靶基因失效的同源对照靶细胞，这提供了理想的对照和纠正非特异性效应的方法。表达不可裂解的膜结合 TNF 的新型效应细胞和靶细胞已被用于定量用英夫利昔单抗治疗的克罗恩病患者血清中的 ADCC 活性，并将 ADCC 活性与药物水平相关联。
