Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Nat Med. 2010 Sep;16(9):1042-7. doi: 10.1038/nm.2205. Epub 2010 Aug 29.
Neutrophils have key roles in modulating the immune response. We present a robust methodology for rapidly isolating neutrophils directly from whole blood with 'on-chip' processing for mRNA and protein isolation for genomics and proteomics. We validate this device with an ex vivo stimulation experiment and by comparison with standard bulk isolation methodologies. Last, we implement this tool as part of a near-patient blood processing system within a multi-center clinical study of the immune response to severe trauma and burn injury. The preliminary results from a small cohort of subjects in our study and healthy controls show a unique time-dependent gene expression pattern clearly demonstrating the ability of this tool to discriminate temporal transcriptional events of neutrophils within a clinical setting.
中性粒细胞在调节免疫反应中起着关键作用。我们提出了一种从全血中快速分离中性粒细胞的稳健方法,该方法具有“片上”处理功能,可用于基因组学和蛋白质组学中的 mRNA 和蛋白质分离。我们通过体外刺激实验和与标准批量分离方法的比较来验证该设备。最后,我们将该工具作为多中心严重创伤和烧伤患者免疫反应临床研究中患者床边血液处理系统的一部分。我们研究中小队列患者和健康对照者的初步结果显示出独特的时间依赖性基因表达模式,清楚地表明该工具能够在临床环境中区分中性粒细胞的时间转录事件。