流式细胞术检测幽门螺杆菌相关性胃炎胃上皮及浸润黏膜淋巴细胞中肿瘤坏死因子相关凋亡诱导配体及其受体
Flow cytometric measurement of tumor necrosis factor-related apoptosis-inducing ligand and its receptors in gastric epithelium and infiltrating mucosal lymphocytes in Helicobacter pylori-associated gastritis.
作者信息
Koyama Shohei
机构信息
Department of Internal Medicine, Institute of Clinical Medicine, University of Tsukuba, Tsukuba, Japan.
出版信息
J Gastroenterol Hepatol. 2003 Jul;18(7):763-70. doi: 10.1046/j.1440-1746.2003.03055.x.
BACKGROUND
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptors have recently been reported to be responsible for apoptotic signaling molecules. However, little is known about TRAIL-mediated apoptosis in the human glandular stomach.
METHODS
Biopsies from 66 patients (28 Helicobacter pylori-negative, 38 H. pylori-positive) were investigated for phenotypic distribution of TRAIL and its receptors DR4/DR5 and DcR2 on mucosal epithelium, and infiltrating mucosal lymphocytes using flow cytometry. Apoptosis of the cells was examined by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick and labeling (TUNEL). In addition, the phenotypic distribution of CD antigens on infiltrating major T- and B-lymphocytes were determined.
RESULTS
Membrane-bound TRAIL and its receptors were constitutively expressed in all patients with high levels in the gastric mucosal epithelium. In particular, these protein molecules were overexpressed in mucosal lymphocytes coupled with increased proportions of CD19+ B cells, and CD3+ T cells bearing CD8+CD11b- and CD4+CD62L- surface phenotypes in H. pylori-positive gastric mucosa. The frequencies of apoptotic epithelium and infiltrating lymphocytes in H. pylori-associated gastritis were significantly greater than those of H. pylori-negative normal mucosa (P < 0.01).
CONCLUSION
The present findings show that flow cytometric analysis is useful for detection of membrane-bound TRAIL and its receptors in gastric epithelium and infiltrating mucosal lymphocytes.
背景
肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其受体最近被报道为凋亡信号分子。然而,关于TRAIL介导的人胃腺凋亡知之甚少。
方法
对66例患者(28例幽门螺杆菌阴性,38例幽门螺杆菌阳性)的活检组织进行研究,采用流式细胞术检测TRAIL及其受体DR4/DR5和DcR2在黏膜上皮以及浸润性黏膜淋巴细胞上的表型分布。通过末端脱氧核苷酸转移酶(TdT)介导的dUTP缺口末端标记法(TUNEL)检测细胞凋亡情况。此外,还测定了浸润的主要T淋巴细胞和B淋巴细胞上CD抗原的表型分布。
结果
膜结合型TRAIL及其受体在所有患者中均有组成性表达,在胃黏膜上皮中表达水平较高。特别是,这些蛋白质分子在黏膜淋巴细胞中过度表达,同时在幽门螺杆菌阳性胃黏膜中,CD19+B细胞以及具有CD8+CD11b-和CD4+CD62L-表面表型的CD3+T细胞比例增加。幽门螺杆菌相关性胃炎中凋亡上皮细胞和浸润淋巴细胞的频率显著高于幽门螺杆菌阴性的正常黏膜(P<0.01)。
结论
本研究结果表明,流式细胞术分析对于检测胃上皮和浸润性黏膜淋巴细胞中的膜结合型TRAIL及其受体很有用。