Tvedt Tor Henrik Anderson, Rye Kristin Paulsen, Reikvam Håkon, Brenner Annette K, Bruserud Øystein
Section of Hematology, Department of Medicine, Haukeland University Hospital, Norway.
Institute of Clinical Science, Section of Hematology, University of Bergen, Norway.
J Immunol Methods. 2015 Mar;418:19-28. doi: 10.1016/j.jim.2015.01.006. Epub 2015 Jan 28.
Cytokines, soluble adhesion molecules and metalloproteinases can be detected in human serum or plasma samples. Such systemic levels are widely used as biomarkers in epidemiological and clinical studies.
We prepared serum samples and three types of plasma samples (EDTA, heparin, citric acid) from 20 healthy individuals. The levels of 31 cytokines, four soluble adhesion molecules and eight matrix metalloproteinases were analyzed by Luminex technology.
Most mediators showed detectable levels in both plasma and serum. Several mediators that can be released by platelets showed increased serum levels, especially CCL5 and CD40L, but for the other mediators the serum levels did not correlate with peripheral blood platelet counts and for these last mediators serum and plasma levels often showed strong correlations. The use of bivalirudin for anticoagulation significantly increased and citric acid combined with platelet inhibitors (ticagrelor, acetylsalicylic acid plus prostaglandin E2) did not alter plasma levels of platelet-store mediators compared with citric acid alone. The impact of sample preparation differed between mediators; for many mediators strong correlations were seen between serum and plasma levels even when absolute levels differed. Soluble adhesion molecule levels showed only minor differences between samples. Unsupervised hierarchical clustering suggested that the effect of sampling/preparation was strongest for serum and heparin plasma samples.
Careful standardization of sample preparation is usually necessary when analyzing systemic mediator levels, and differences caused by sample preparation should be considered as a possible explanation if studies show conflicting results.
细胞因子、可溶性黏附分子和金属蛋白酶可在人血清或血浆样本中检测到。这些全身性水平在流行病学和临床研究中被广泛用作生物标志物。
我们从20名健康个体中制备了血清样本和三种血浆样本（乙二胺四乙酸、肝素、柠檬酸）。通过Luminex技术分析了31种细胞因子、四种可溶性黏附分子和八种基质金属蛋白酶的水平。
大多数介质在血浆和血清中均显示出可检测到的水平。几种可由血小板释放的介质血清水平升高，尤其是CCL5和CD40L，但对于其他介质，血清水平与外周血血小板计数无关，并且对于这些最后介质，血清和血浆水平通常显示出强相关性。与单独使用柠檬酸相比，使用比伐卢定进行抗凝显著增加，而柠檬酸与血小板抑制剂（替格瑞洛、乙酰水杨酸加前列腺素E2）联合使用并未改变血小板储存介质的血浆水平。介质之间样本制备的影响有所不同；对于许多介质，即使绝对水平不同，血清和血浆水平之间也存在强相关性。可溶性黏附分子水平在样本之间仅显示出微小差异。无监督层次聚类表明，采样/制备的影响在血清和肝素血浆样本中最为明显。
在分析全身性介质水平时，通常需要仔细标准化样本制备，如果研究结果相互矛盾，样本制备引起的差异应被视为一种可能的解释。
