Department of Biological Sciences, Columbia University, Northwest Corner Building, 12th Floor, MC 4846, 550 West 120(th) Street, New York, NY 10027, USA.
HelmholtzZentrum München, German Research Center for Environmental Health (GmbH), Assay Development and Screening Platform, Institute for Molecular Toxicology and Pharmacology, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
Cell Rep. 2020 Mar 10;30(10):3411-3423.e7. doi: 10.1016/j.celrep.2020.02.049.
Ferroptosis is a type of regulated cell death driven by the iron-dependent accumulation of oxidized polyunsaturated fatty acid-containing phospholipids. There is no reliable way to selectively stain ferroptotic cells in tissue sections to characterize the extent of ferroptosis in animal models or patient samples. We address this gap by immunizing mice with membranes from lymphoma cells treated with the ferroptosis inducer piperazine erastin and screening ∼4,750 of the resulting monoclonal antibodies generated for their ability to selectively detect cells undergoing ferroptosis. We find that one antibody, 3F3 ferroptotic membrane antibody (3F3-FMA), is effective as a selective ferroptosis-staining reagent. The antigen of 3F3-FMA is identified as the human transferrin receptor 1 protein (TfR1). We validate this finding with several additional anti-TfR1 antibodies and compare them to other potential ferroptosis-detecting reagents. We find that anti-TfR1 and anti-malondialdehyde adduct antibodies are effective at staining ferroptotic tumor cells in multiple cell culture and tissue contexts.
铁死亡是一种受铁依赖性氧化多不饱和脂肪酸含磷脂积累驱动的调节性细胞死亡。目前还没有可靠的方法可以在组织切片中选择性地上染铁死亡细胞，以表征动物模型或患者样本中的铁死亡程度。我们通过用铁死亡诱导剂哌嗪 erastin 处理的淋巴瘤细胞的膜免疫小鼠，并筛选大约 4750 个由此产生的单克隆抗体，以评估它们选择性检测发生铁死亡的细胞的能力，从而解决了这一空白。我们发现一种抗体，3F3 铁死亡膜抗体（3F3-FMA），可作为一种有效的选择性铁死亡染色试剂。3F3-FMA 的抗原被鉴定为人类转铁蛋白受体 1 蛋白（TfR1）。我们用其他几种抗 TfR1 抗体对此进行了验证，并将其与其他潜在的铁死亡检测试剂进行了比较。我们发现抗 TfR1 和抗丙二醛加合物抗体可有效地在多种细胞培养和组织环境中对铁死亡的肿瘤细胞进行染色。
