Quantitative whole-cell MALDI-TOF MS fingerprints distinguishes human monocyte sub-populations activated by distinct microbial ligandsReportar como inadecuado




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BMC Biotechnology

, 15:24

First Online: 11 April 2015Received: 30 October 2014Accepted: 30 March 2015DOI: 10.1186-s12896-015-0140-1

Cite this article as: Portevin, D., Pflüger, V., Otieno, P. et al. BMC Biotechnol 2015 15: 24. doi:10.1186-s12896-015-0140-1

Abstract

BackgroundConventionally, human monocyte sub-populations are classified according to surface marker expression into classical CD14CD16, intermediate CD14CD16 and non-classical CD14CD16 lineages. The involvement of non-classical monocytes, also referred to as proinflammatory monocytes, in the pathophysiology of diseases including diabetes mellitus, atherosclerosis or Alzheimer’s disease is well recognized. The development of novel high-throughput methods to capture functional states within the different monocyte lineages at the whole cell proteomic level will enable real time monitoring of disease states.

ResultsWe isolated and characterized pan- monocytes, mostly composed of classical CD16 monocytes, versus autologous CD16 subpopulations from the blood of healthy human donors n = 8 and compared their inflammatory properties in response to lipopolysaccharides and M.tuberculosis antigens by multiplex cytokine profiling. Following resting and in vitro antigenic stimulation, cells were recovered and subjected to whole-cell mass spectrometry analysis. This approach identified the specific presence-absence of m-z peaks and therefore potential biomarkers that can discriminate pan-monocytes from their CD16 counterparts. Furthermore, we found that semi-quantitative data analysis could capture the subtle proteome changes occurring upon microbial stimulation that differentiate resting, from lipopolysaccharides or M. tuberculosis stimulated monocytic samples.

ConclusionsWhole-cell mass spectrometry fingerprinting could efficiently distinguish monocytic sub-populations that arose from a same hematopoietic lineage. We also demonstrate for the first time that mass spectrometry signatures can monitor semi-quantitatively specific activation status in response to exogenous stimulation. As such, this approach stands as a fast and efficient method for the applied immunology field to assess the reactivity of potentially any immune cell types that may sustain health or promote related inflammatory diseases.

KeywordsWhole-cell mass spectrometry MALDIquant Fingerprints Monocyte CD14 CD16 MDC8 Electronic supplementary materialThe online version of this article doi:10.1186-s12896-015-0140-1 contains supplementary material, which is available to authorized users.

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Autor: Damien Portevin - Valentin Pflüger - Patricia Otieno - René Brunisholz - Guido Vogel - Claudia Daubenberger

Fuente: https://link.springer.com/







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