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Gene Expression Profiles from Peripheral Blood Mononuclear Cells Are Sensitive to Short Processing Delays

机译：外周血单个核细胞的基因表达谱对短加工延迟敏感。

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In the analysis of peripheral blood gene expression, timely processing of samples is essential to ensure that measurements reflect in vivo biology, rather than ex vivo sample processing variables. The effect of processing delays on global gene expression patterns in peripheral blood mononuclear cells (PBMCs) was assessed by isolating and stabilizing PBMC-derived RNA from 3 individuals either immediately after phlebotomy or after a 4 h delay. RNA was labeled using NuGEN Ovation labeling and probed using the Affymetrix HG U133 Plus 2.0 GeneChip®. Comparison of gene expression levels (≥2-fold expression change and P < 0.05) identified 307 probe sets representing genes with increased expression and 46 indicating decreased expression after 4 h. These differentially expressed genes include many that are important to inflammatory, immunologic, and cancer pathways. Among others, CCR2, CCR5, TLR10, CD180, and IL-16 have decreased expression, whereas VEGF, IL8, SOCS2, SOCS3, CD69, and CD83 have increased expression after a 4 h processing delay. The trends in expression patterns associated with delayed processing were also apparent in an independent set of 276 arrays of RNA from human PBMC samples with varying processing times. These data indicate that the time between sample acquisition, initiation of processing, and when the RNA is stabilized should be a prime consideration when designing protocols for translational studies involving PBMC gene expression analysis.
机译：在分析外周血基因表达时，及时处理样品对于确保测量结果反映体内生物学而不是离体样品处理变量至关重要。通过在放血后或延迟4小时后分离并稳定来自3个个体的PBMC衍生的RNA，评估了处理延迟对外周血单核细胞（PBMC）全局基因表达模式的影响。使用NuGEN Ovation标记法标记RNA，并使用Affymetrix HG U133 Plus 2.0 GeneChip ®进行探测。比较基因表达水平（≥2倍表达变化，P <0.05），鉴定出307个探针组，代表4个h后表达增强的基因和46个表达水平降低的探针。这些差异表达的基因包括许多对炎症，免疫和癌症途径重要的基因。其中，CCR2，CCR5，TLR10，CD180和IL-16的表达降低，而VEGF，IL8，SOCS2，SOCS3，CD69和CD83的表达在经过4小时的处理延迟后增加。与一组延迟加工相关的表达模式的趋势在来自不同处理时间的人PBMC样品的276个RNA阵列的独立集合中也很明显。这些数据表明，在设计涉及PBMC基因表达分析的翻译研究方案时，应首先考虑样品获取，处理开始以及RNA稳定之间的时间。

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