Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology (ETH Zürich), Vladimir-Prelog-Weg 4, CH-8093, Zürich, Switzerland.
Sci Rep. 2020 Feb 24;10(1):3234. doi: 10.1038/s41598-020-60182-4.
Engineered cytokine products represent a growing class of therapeutic proteins which need to be tested for biological activity at various stages of pharmaceutical development. In most cases, dedicated biological assays are established for different products, in a process that can be time-consuming and cumbersome. Here we describe the development and implementation of a universal cell-based reporter system for various classes of immunomodulatory proteins. The novel system capitalizes on the fact that the signaling of various types of pro-inflammatory agents (e.g., cytokines, chemokines, Toll-like receptor agonists) may involve transcriptional activation by NF-κB. Using viral transduction, we generated stably-transformed cell lines of B or T lymphocyte origin and compared the new reporter cell lines with conventional bioassays. The experimental findings with various interleukins and with members of the TNF superfamily revealed that the newly-developed "universal" bioassay method yielded bioactivity data which were comparable to the ones obtained with dedicated conventional methods. The engineered cell lines with reporters for NF-κB were tested with several antibody-cytokine fusions and may be generally useful for the characterization of novel immunomodulatory products. The newly developed methodology also revealed a mechanism for cytokine potentiation, based on the antibody-mediated clustering of TNF superfamily members on tumor-associated extracellular matrix components.
工程细胞因子产品代表了一类不断增长的治疗性蛋白，需要在药物开发的各个阶段测试其生物活性。在大多数情况下，针对不同的产品建立了专门的生物学检测方法，这一过程可能既耗时又麻烦。在这里，我们描述了一种用于各种免疫调节蛋白的通用基于细胞的报告系统的开发和实施。该新型系统利用了这样一个事实，即各种类型的促炎剂（例如细胞因子、趋化因子、Toll 样受体激动剂）的信号转导可能涉及 NF-κB 的转录激活。我们通过病毒转导生成了 B 或 T 淋巴细胞来源的稳定转化细胞系，并将新的报告细胞系与传统的生物测定进行了比较。使用各种白细胞介素和 TNF 超家族成员进行的实验发现，新开发的“通用”生物测定方法产生的生物活性数据与使用专门的传统方法获得的数据相当。用于 NF-κB 的报告基因的工程细胞系与几种抗体-细胞因子融合体进行了测试，可能对新型免疫调节产品的特性具有普遍意义。新开发的方法还揭示了一种基于 TNF 超家族成员在肿瘤相关细胞外基质成分上的抗体介导聚集的细胞因子增强机制。
