ABSTRACT

Therapeutic antibodies sometimes elicit anti-drug antibodies (ADAs) that can affect efficacy and safety. Engineered antibodies that contain artificial amino acid sequences are potentially highly immunogenic, but this is currently difficult to predict. Therefore, it is important to efficiently assess immunogenicity during the development of complex antibody-based formats. Here, we present an in vitro peripheral blood mononuclear cell-based assay that can be used to assess immunogenicity potential within 3 days. This method involves examining the frequency and function of interleukin (IL)-2-secreting CD4⁺ T cells induced by therapeutic antibodies. IL-2-secreting CD4⁺ T cells seem to be functionally relevant to the immunogenic potential due to their proliferative activity and the expression of several cytokines. The rates of the donors responding to low and high immunogenic proteins, mAb1, and keyhole limpet hemocyanin were 1.3% and 93.5%, respectively. Seven antibodies with known rates of immunogenicity (etanercept, emicizumab, abciximab, romosozumab, blosozumab, humanized anti-human A33 antibody, and bococizumab) induced responses in 1.9%, 3.8%, 6.4%, 10.0%, 29.2%, 43.8%, and 89.5% of donors, respectively. These data are comparable with ADA incidences in clinical settings. Our results show that this assay can contribute to the swift assessment and mechanistic understanding of the immunogenicity of therapeutic antibodies.

摘要

治疗性抗体有时会引发抗药物抗体 (ADA)，从而影响疗效和安全性。含有人工氨基酸序列的工程抗体具有潜在的高免疫原性，但这目前很难预测。因此，在复杂的基于抗体的形式的开发过程中，有效评估免疫原性非常重要。在这里，我们提出了一种基于体外外周血单核细胞的检测方法，可用于在 3 天内评估免疫原性潜力。^{该方法涉及检查治疗性抗体诱导的白细胞介素 (IL)-2 分泌 CD4 +} T 细胞的频率和功能。由于其增殖活性和多种细胞因子的表达，分泌 IL-2 的 CD4 ^{+ T 细胞似乎在功能上与免疫原性潜力相关。}供体对低免疫原性蛋白、mAb1 和匙孔血蓝蛋白的反应率分别为 1.3% 和 93.5%。七种具有已知免疫原性的抗体（依那西普、艾美珠单抗、阿昔单抗、罗莫佐单抗、blosozumab、人源化抗人 A33 抗体和博可珠单抗）诱导的应答分别为 1.9%、3.8%、6.4%、10.0%、29.2%、43.8% 和捐赠者的比例分别为 89.5%。这些数据与临床环境中 ADA 的发生率具有可比性。我们的结果表明，该测定有助于快速评估和理解治疗性抗体的免疫原性。