Institute of Analytical Chemistry, University of Vienna, Austria.
J Proteomics. 2012 Dec 5;76 Spec No.(5):150-62. doi: 10.1016/j.jprot.2012.07.012. Epub 2012 Jul 16.
Proteome profiling is the method of choice to identify marker proteins whose expression may be characteristic for certain diseases. The formation of such marker proteins results from disease-related pathophysiologic processes. In healthy individuals, peripheral blood mononuclear cells (PBMCs) circulate in a quiescent cell state monitoring potential immune-relevant events, but have the competence to respond quickly and efficiently in an inflammatory manner to any invasion of potential pathogens. Activation of these cells is most plausibly accompanied by characteristic proteome alterations. Therefore we investigated untreated and inflammatory activated primary human PBMCs by proteome profiling using a 'top down' 2D-PAGE approach in addition to a 'bottom up' LC-MS/MS-based shotgun approach. Furthermore, we purified primary human T-cells and monocytes and activated them separately. Comparative analysis allowed us to characterize a robust proteome signature including NAMPT and PAI2 which indicates the activation of PBMCs. The T-cell specific inflammation signature included IRF-4, GBP1 and the previously uncharacterized translation product of GBP5; the corresponding monocyte signature included PDCD5, IL1RN and IL1B. The involvement of inflammatory activated PBMCs in certain diseases as well as the responsiveness of these cells to anti-inflammatory drugs may be evaluated by quantification of these marker proteins. This article is part of a Special Issue entitled: Integrated omics.
蛋白质组谱分析是鉴定可能与某些疾病相关的标记蛋白的首选方法。这些标记蛋白的形成源于与疾病相关的病理生理过程。在健康个体中，外周血单核细胞（PBMC）处于静止状态循环，监测潜在的免疫相关事件，但具有快速、高效地以炎症方式对任何潜在病原体入侵做出反应的能力。这些细胞的激活最有可能伴随着特征性的蛋白质组改变。因此，我们使用“自上而下”的 2D-PAGE 方法和“自下而上”的基于 LC-MS/MS 的鸟枪法对未经处理和炎症激活的原代人 PBMC 进行了蛋白质组谱分析。此外，我们还纯化了原代人 T 细胞和单核细胞并分别对其进行了激活。比较分析使我们能够表征一个包括 NAMPT 和 PAI2 的稳健蛋白质组特征，这表明 PBMC 被激活。T 细胞特异性炎症特征包括 IRF-4、GBP1 和 GBP5 的以前未表征的翻译产物；相应的单核细胞特征包括 PDCD5、IL1RN 和 IL1B。通过定量这些标记蛋白，可以评估炎症激活的 PBMC 在某些疾病中的参与以及这些细胞对抗炎药物的反应性。本文是一个题为“综合组学”的特刊的一部分。
