Laboratory of Experimental Cardiology, University Medical Centre Utrecht, Utrecht, The Netherlands.
PLoS One. 2013;8(4):e60467. doi: 10.1371/journal.pone.0060467. Epub 2013 Apr 3.
Toll-Like Receptor (TLR) -2 and -4 expression and TLR-induced cytokine response of inflammatory cells are related to atherogenesis and atherosclerotic plaque progression. We examined whether immediate TLR induced changes in CD11b and L-selectin (CD62L) expression are able to discriminate the presence and severity of atherosclerotic disease by exploring single dose whole blood TLR stimulation and detailed dose-response curves. Blood samples were obtained from 125 coronary artery disease (CAD) patients and 28 controls. CD11b and L-selectin expression on CD14+ monocytes was measured after whole blood stimulation with multiple concentrations of the TLR4 ligand LPS (0.01-10 ng/ml) and the TLR2 ligand P3C (0.5-500 ng/ml). Subsequently, dose-response curves were created and the following parameters were calculated: hillslope, EC50, area under the curve (AUC) and delta. These parameters provide information about the maximum response following activation, as well as the minimum trigger required to induce activation and the intensity of the response. CAD patients showed a significantly higher L-selectin, but not CD11b response to TLR ligation than controls after single dose stimulations as well as significant differences in the hillslope and EC50 of the dose-response curves. Within the CAD patient group, dose-response curves of L-selectin showed significant differences in the presence of hypertension, dyslipidemia, coronary occlusion and degree of stenosis, whereas CD11b expression had the strongest discriminating power after single dose stimulation. In conclusion, single dose stimulations and dose-response curves of CD11b and L-selectin expression after TLR stimulation provide diverse but limited information about atherosclerotic disease severity in stable angina patients. However, both single dose stimulation and dose-response curves of LPS-induced L-selectin expression can discriminate between controls and CAD patients.
Toll 样受体 (TLR)-2 和 -4 的表达以及 TLR 诱导的炎症细胞细胞因子反应与动脉粥样硬化形成和动脉粥样硬化斑块进展有关。我们通过研究单次全血 TLR 刺激和详细的剂量反应曲线,来检测 TLR 诱导的 CD11b 和 L-选择素(CD62L)表达的即时变化是否能够通过探索单一剂量全血 TLR 刺激和详细的剂量反应曲线来区分动脉粥样硬化疾病的存在和严重程度。从 125 名冠心病 (CAD) 患者和 28 名对照者中采集血样。用不同浓度的 TLR4 配体 LPS(0.01-10ng/ml)和 TLR2 配体 P3C(0.5-500ng/ml)刺激全血后,测量 CD14+单核细胞上的 CD11b 和 L-选择素表达。随后,创建剂量反应曲线,并计算以下参数:坡度、EC50、曲线下面积 (AUC) 和 delta。这些参数提供了关于激活后最大反应以及激活所需的最小触发和反应强度的信息。与单次刺激后对照组相比,CAD 患者在 TLR 连接后显示出更高的 L-选择素但不是 CD11b 反应,并且在剂量反应曲线的坡度和 EC50 方面也存在显著差异。在 CAD 患者组中,L-选择素的剂量反应曲线在高血压、血脂异常、冠状动脉阻塞和狭窄程度存在差异,而 CD11b 表达在单次刺激后具有最强的区分能力。总之,TLR 刺激后 CD11b 和 L-选择素表达的单次剂量刺激和剂量反应曲线为稳定型心绞痛患者提供了关于动脉粥样硬化严重程度的多样化但有限的信息。然而,LPS 诱导的 L-选择素表达的单次剂量刺激和剂量反应曲线都可以区分对照组和 CAD 患者。