In-Depth Profiling of the Peripheral Blood Mononuclear Cells Proteome for Clinical Blood Proteomics

Abstract

Peripheral blood mononuclear cells (PBMCs) are an easy accessible cellular part of the blood organ and, along with platelets, represent the only site of active gene expression in blood. These cells undergo immunophenotypic changes in various diseases and represent a peripheral source of monitoring gene expression and posttranslational modifications relevant to many diseases. Little is known about the source of many blood proteins and we hypothesise that release from PBMCs through active and passive mechanisms may account for a substantial part of the plasma proteome. The use of state-of-the-art proteomic profiling methods in PBMCs will enable minimally invasive monitoring of disease progression or response to treatment and discovery of biomarkers. To achieve this goal, detailed mapping of the PBMC proteome using a sensitive, robust, and quantitative methodological setup is required. We have applied an indepth gel-free proteomics approach using tandem mass tags (TMT), unfractionated and SCX fractionated PBMC samples, and LC-MS/MS with various modulations. This study represents a benchmark in deciphering the PBMC proteome as we provide a deep insight by identifying 4129 proteins and 25503 peptides. The identified proteome defines the scope that enables PBMCs to be characterised as cellular major biomarker pool within the blood organ.

T cells from hemophilia A subjects recognize the same HLA-restricted FVIII epitope with a narrow TCR repertoire

“…Intracellular synthesis of a partial FVIII and of the FVIII B proteins has been reported in PBMCs and liver tissue from intron-22-inversion HA subjects. 62,63 However, recent proteomics studies have not confirmed FVIII or FVIII B expression in PBMCs or in thymus peptides associated with HLA-DR. 64,65 The isolation reported herein of FVIII-C2-specific T-effector clones and lines from a severe HA subject whose F8 mutation did not affect the F8B region demonstrates directly that the FVIII-C2 domain is a target for T cells in HA subjects and thereby argues against widespread central tolerance to this domain.…”

Up-regulation of MSH2, XRCC1 and ATM genes in patients with type 2 diabetes and coronary artery disease

“…These cells suffer from DNA lesions in CAD patients but little is known about its underlying molecular events. Furthermore, their availability makes them a suitable source of noninvasive predictive biomarkers in clinical studies [20].…”

Downregulated microRNA-155 expression in peripheral blood mononuclear cells of type 2 diabetic patients is not correlated with increased inflammatory cytokine production

“…PBMCs contain different cell types including, T, B, natural killer (NK) and dendritic cells. These cells play important roles in the immune system monitoring, and immune-relevant events [9]. These cells undergo immunophenotypic changes in various diseases and represent a peripheral source of monitoring gene expression and posttranslational modifications relevant to many diseases.…”