Mitsuhashi Shuji, Feldbrügge Linda, Csizmadia Eva, Mitsuhashi Masato, Robson Simon C, Moss Alan C
*Division of Gastroenterology, Beth Israel Deaconess Medical Center, Boston, Massachusetts; and †Hitachi Chemical Co. America, Ltd., Irvine, California.
Inflamm Bowel Dis. 2016 Jul;22(7):1587-95. doi: 10.1097/MIB.0000000000000840.
Extracellular vesicles (EVs) are membrane-enclosed particles released by cells as a means of intercellular communication. They are potential novel biomarkers, as they are readily isolated from body fluids, and their composition reflects disease pathways. Whether these particles are released from sites of intestinal inflammation in inflammatory bowel disease (IBD) has not previously been determined.
EVs were isolated by ultracentrifugation of colonic luminal fluid aspirates and characterized according to surface proteins, and constituent mRNA and proteins. The effects of EVs on colonic epithelial cells and macrophages in culture were assessed at the transcriptional, translational, and functional levels.
Intestinal luminal aspirates contained abundant EVs, at a mean concentration of 4.3 × 10 particles/mL and with a mean diameter of 146 nm. EVs from patients with IBD with a high endoscopic score (≥1) contained significantly higher mRNA and protein levels of interleukin 6 (IL-6), IL-8, IL-10, and tumor necrosis factor α than EVs from healthy controls. EVs were absorbed by cultured colonic epithelial cells, leading to an increased translation of IL-8 protein by recipient cells when treated with EVs from patients with IBD. EVs and EV-treated epithelial cells induced migration of a significantly greater number of macrophages than epithelial cells alone.
EVs shed from sites of intestinal inflammation in patients with IBD have a distinct mRNA and protein profile from those of healthy individuals. These EVs have proinflammatory effects on the colonic epithelium, in vitro. Their stability in luminal samples and their mRNA and protein content identify them as a potential fecal biomarker that reflects mucosal inflammatory pathways.
细胞外囊泡(EVs)是细胞释放的膜包裹颗粒,作为细胞间通讯的一种方式。它们是潜在的新型生物标志物,因为它们易于从体液中分离出来,并且其组成反映疾病途径。此前尚未确定这些颗粒是否从炎症性肠病(IBD)的肠道炎症部位释放。
通过对结肠腔液吸出物进行超速离心分离EVs,并根据表面蛋白、组成性mRNA和蛋白质进行表征。在转录、翻译和功能水平评估EVs对培养的结肠上皮细胞和巨噬细胞的影响。
肠道腔吸出物中含有丰富的EVs,平均浓度为4.3×10颗粒/毫升,平均直径为146纳米。内镜评分高(≥1)的IBD患者的EVs中白细胞介素6(IL-6)、IL-8、IL-10和肿瘤坏死因子α的mRNA和蛋白质水平显著高于健康对照的EVs。EVs被培养的结肠上皮细胞吸收,当用IBD患者的EVs处理时,受体细胞中IL-8蛋白的翻译增加。与单独的上皮细胞相比,EVs和经EV处理的上皮细胞诱导迁移的巨噬细胞数量显著更多。
IBD患者肠道炎症部位脱落的EVs与健康个体的EVs具有不同的mRNA和蛋白质谱。这些EVs在体外对结肠上皮具有促炎作用。它们在腔样本中的稳定性及其mRNA和蛋白质含量使其成为反映黏膜炎症途径的潜在粪便生物标志物。