Bethel Kelly, Luttgen Madelyn S, Damani Samir, Kolatkar Anand, Lamy Rachelle, Sabouri-Ghomi Mohsen, Topol Sarah, Topol Eric J, Kuhn Peter
Scripps Clinic, Department of Pathology, 10660 North Torrey Pines Road, MC211C, La Jolla, CA 92037, USA.
Phys Biol. 2014 Feb;11(1):016002. doi: 10.1088/1478-3975/11/1/016002. Epub 2014 Jan 9.
Elevated levels of circulating endothelial cells (CECs) occur in response to various pathological conditions including myocardial infarction (MI). Here, we adapted a fluid phase biopsy technology platform that successfully detects circulating tumor cells in the blood of cancer patients (HD-CTC assay), to create a high-definition circulating endothelial cell (HD-CEC) assay for the detection and characterization of CECs. Peripheral blood samples were collected from 79 MI patients, 25 healthy controls and six patients undergoing vascular surgery (VS). CECs were defined by positive staining for DAPI, CD146 and von Willebrand Factor and negative staining for CD45. In addition, CECs exhibited distinct morphological features that enable differentiation from surrounding white blood cells. CECs were found both as individual cells and as aggregates. CEC numbers were higher in MI patients compared with healthy controls. VS patients had lower CEC counts when compared with MI patients but were not different from healthy controls. Both HD-CEC and CellSearch® assays could discriminate MI patients from healthy controls with comparable accuracy but the HD-CEC assay exhibited higher specificity while maintaining high sensitivity. Our HD-CEC assay may be used as a robust diagnostic biomarker in MI patients.
循环内皮细胞(CEC)水平升高会在包括心肌梗死(MI)在内的各种病理状况下出现。在此,我们采用了一种液相活检技术平台(该平台已成功检测出癌症患者血液中的循环肿瘤细胞,即高清循环肿瘤细胞(HD-CTC)检测法),来创建一种用于检测和鉴定CEC的高清循环内皮细胞(HD-CEC)检测法。从79名心肌梗死患者、25名健康对照者以及6名接受血管手术(VS)的患者中采集外周血样本。CEC通过DAPI、CD146和血管性血友病因子的阳性染色以及CD45的阴性染色来定义。此外,CEC呈现出独特的形态特征,使其能够与周围的白细胞区分开来。CEC既以单个细胞形式存在,也以聚集体形式存在。与健康对照者相比,心肌梗死患者的CEC数量更多。与心肌梗死患者相比,血管手术患者的CEC计数较低,但与健康对照者无差异。HD-CEC检测法和CellSearch®检测法都能以相当的准确性区分心肌梗死患者和健康对照者,但HD-CEC检测法在保持高灵敏度的同时表现出更高的特异性。我们的HD-CEC检测法可用作心肌梗死患者一种可靠的诊断生物标志物。