The impact of blood-processing time on the proteome of human peripheral blood mononuclear cells
| Autor: | Amanda Caroline Camillo-Andrade, Saloe Bispo, Marlon D.M. Santos, Juliana de Saldanha da Gama Fischer, Fábio C. S. Nogueira, Bernardo Bonilauri, Nilson Ivo Tonin Zanchin, Paulo C. Carvalho |
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| Rok vydání: | 2021 |
| Předmět: |
Adult
Male Proteomics 0301 basic medicine Granulocyte activation Proteome Biophysics Granulocyte Biochemistry Peripheral blood mononuclear cell Mass Spectrometry Analytical Chemistry Young Adult 03 medical and health sciences 0302 clinical medicine Leukocyte degranulation medicine Humans Protein Interaction Maps Molecular Biology Chemistry Blood Proteins Myeloid leukocyte activation Cell biology Gene Ontology 030104 developmental biology Mitochondrial respiratory chain medicine.anatomical_structure 030220 oncology & carcinogenesis Leukocytes Mononuclear Neutrophil degranulation Cell activation Chromatography Liquid |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 1869:140581 |
| ISSN: | 1570-9639 |
| DOI: | 10.1016/j.bbapap.2020.140581 |
| Popis: | Human peripheral blood mononuclear cells (PBMC) are key to several diagnostics assays and basic science research. Blood pre-analytical variations that occur before obtaining the PBMC fraction can significantly impact the assays results, including viability, composition, integrity, and gene expression changes of immune cells. With this as motivation, we performed a quantitative shotgun proteomics analysis using Isobaric Tag for Relative and Absolute Quantitation (iTRAQ 8plex) labeling to compare PBMC obtained from 24 h-stored blood at room temperature versus freshly isolated. We identified a total of 3195 proteins, of which 245 were differentially abundant (101 upregulated and 144 downregulated). Our results revealed enriched pathways of downregulated proteins related to exocytosis, localization, vesicle-mediated transport, cell activation, and secretion. In contrast, pathways related to exocytosis, neutrophil degranulation and activation, granulocyte activation, leukocyte degranulation, and myeloid leukocyte activation involved in immune response were enriched in upregulated proteins, which may indicate probable granulocyte contamination and activation due to blood storage time and temperature. Examples of upregulated proteins in the 24 h-PBMC samples are CAMP, S100A8, LTA4H, RASAL3, and S100A6, which are involved in an adaptive immune system and antimicrobial activity, proinflammatory mediation, aminopeptidase activities, and naive T cells survival. Moreover, examples of downregulated proteins are NDUFA5, TAGLN2, H3C1, TUBA8, and CCT2 that are related to the cytoskeleton, cell junction, mitochondrial respiratory chain. In conclusion, the delay in blood-processing time directly impacts the proteomic profile of human PBMC, possibly through granulocyte contamination and activation. |
| Databáze: | OpenAIRE |
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