Equipe labellisée par la Ligue contre le cancer, Université de Paris, Sorbonne Université, INSERM UMR1138, Centre de Recherche des Cordeliers, Paris, France; Metabolomics and Cell Biology Platforms, Gustave Roussy, Villejuif, France.
Equipe labellisée par la Ligue contre le cancer, Université de Paris, Sorbonne Université, INSERM UMR1138, Centre de Recherche des Cordeliers, Paris, France; Metabolomics and Cell Biology Platforms, Gustave Roussy, Villejuif, France; Université Paris Sud, Paris Saclay, Faculty of Medicine, Kremlin Bicêtre, France.
Methods Cell Biol. 2022;172:135-143. doi: 10.1016/bs.mcb.2021.12.028. Epub 2022 Jan 17.
The radiochemotherapy- or chemotherapy-induced stimulation of immunogenic cell death (ICD) affecting malignant cells ignites antitumor immune responses that are clinically relevant as they allow to achieve durable responses beyond treatment discontinuation. The mechanistic exploration of ICD and the discovery of agents and interventions that are endowed with the capacity to elicit ICD is of the utmost importance. Here, we describe an assay for the assessment of type I interferon (IFN) production, which is one of the salient features of ICD. Biosensor cells that express GFP under the control of the IFN-inducible MX dynamin like GTPase 1 (MX1) gene promoter are employed, and the fluorescent signal is assessed by automated microscopy. The described workflow is automation-friendly, rendering it compatible with high-throughput screening (HTS) for drug discovery.
放化疗或化疗诱导的免疫原性细胞死亡（ICD）刺激影响恶性细胞，引发抗肿瘤免疫反应，这在临床上具有重要意义，因为它们可以在治疗停止后实现持久的反应。ICD 的机制探索以及发现具有诱导 ICD 能力的药物和干预措施至关重要。在这里，我们描述了一种用于评估 I 型干扰素（IFN）产生的测定法，这是 ICD 的显著特征之一。我们使用表达 GFP 的生物传感器细胞，该 GFP 在 IFN 诱导的肌球蛋白轻链激酶样 GTP 酶 1（MX1）基因启动子的控制下表达，并通过自动显微镜评估荧光信号。所描述的工作流程易于自动化，使其与药物发现的高通量筛选（HTS）兼容。
