Single molecule measurements of tumor necrosis factor α and interleukin-6 in the plasma of patients with Crohn's disease
Single molecule measurements of tumor necrosis factor α and interleukin-6 in the plasma of patients with Crohn's disease
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DOI：
10.1016/j.jim.2011.07.015
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发表时间：
2011-09-30
影响因子：
2.2
通讯作者：
Duffy, David C.
中科院分区：
文献类型：
--
作者：
Song, Linan;Hanlon, David W.;Duffy, David C.
血液中炎性细胞因子的定量测量受到常规免疫测定灵敏度不足的限制。这种局限性阻碍了对慢性炎性疾病中细胞因子浓度的广泛临床监测。我们应用一种灵敏的单分子检测技术来测量克罗恩病（CD）患者在抗TNF-α治疗前后血浆中的TNF-α和IL-6。在开始抗TNF-α治疗前，收集17例CD患者的血浆，并测定每例患者的克罗恩病活动指数（CDAI）。这些患者的一个子集在治疗开始后12周返回进行随访。还收集了来自年龄和性别匹配的对照的血浆。针对TNF-α和IL-6开发数字ELISA，并使用数字ELISA测定这些细胞因子的血浆浓度。TNF-α和IL-6数字ELISA的检测限分别为0.008 pg/mL和0.006 pg/mL。使用数字ELISA和TNF-α的浓度在所有样品中检测两种细胞因子。CD患者血浆IL-6水平分别为（3.6 ± 0.9）pg/mL和（10.9 ± 112）pg/mL。患者和健康对照组的TNF-α水平无显著差异，但患者血浆中的IL-6水平与对照组相比显著升高。治疗后，游离TNF-α和IL-6浓度的平均降低分别为46%和58%。数字ELISA提供了CD人群中所有患者血浆中TNF-α和IL-6浓度的首次定量测量。治疗后细胞因子浓度的变化-由于数字ELISA的高灵敏度，可以量化-可用于监测治疗效果。(C)2011 Elsevier B. V.保留所有权利。
The quantitative measurement of inflammatory cytokines in blood has been limited by insufficient sensitivity of conventional immunoassays. This limitation has prevented the widespread clinical monitoring of cytokine concentrations in chronic inflammatory diseases. We applied a sensitive, single molecule detection technology to measure TNF-alpha and IL-6 in the plasma of patients with Crohn's disease (CD), before and after treatment with anti-TNF-alpha therapy. Plasma from 17 patients with CD was collected prior to initiation of anti-TNF-alpha therapy, and the Crohn's disease activity index (CDAI) was determined for each patient. A sub-set of these patients returned for follow up 12 weeks after treatment started. Plasma from age- and gender-matched controls was also collected. Digital ELISAs were developed for TNF-alpha and IL-6, and the plasma concentrations of these cytokines were determined using digital ELISA. The limits of detection of the TNF-alpha and IL-6 digital ELISAs were 0.008 pg/mL and 0.006 pg/mL, respectively. Both cytokines were detected in all samples using digital ELISA and the concentrations of TNF-alpha. and IL-6 in the plasma of patients with CD were (3.6 +/- 0.9) pg/mL and (10.9 +/- 112) pg/mL, respectively. TNF-alpha levels in patients and healthy controls were not significantly different, but the IL-6 levels in plasma were significantly elevated in patients compared to controls. After therapy, the mean reduction of the concentrations of free TNF-alpha and IL-6 were 46% and 58%, respectively. Digital ELISA provided the first quantitative measurements of TNF-alpha and IL-6 concentrations in the plasma of all patients in a population with CD. The changes in cytokine concentrations after therapy-which could be quantified because of the high sensitivity of digital ELISA-could be used for monitoring therapeutic efficacy. (C) 2011 Elsevier B.V. All rights reserved.