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Issue 6, 2015
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Trace element landscape of resting and activated human neutrophils on the sub-micrometer level

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Abstract

Every infection is a battle for trace elements. Neutrophils migrate first to the infection site and accumulate quickly to high numbers. They fight pathogens by phagocytosis and intracellular toxication. Additionally, neutrophils form neutrophil extracellular traps (NETs) to inhibit extracellular microbes. Yet, neutrophil trace element characteristics are largely unexplored. We investigated unstimulated and phorbol myristate acetate-stimulated neutrophils using synchrotron radiation X-ray fluorescence (SR-XRF) on the sub-micron spatial resolution level. PMA activates pinocytosis, cytoskeletal rearrangements and the release of NETs, all mechanisms deployed by neutrophils to combat infection. By analyzing Zn, Fe, Cu, Mn, P, S, and Ca, not only the nucleus but also vesicular granules were identifiable in the elemental maps. Inductively Coupled Plasma Mass Spectrometry (ICP-MS) revealed a neutrophil-specific composition of Zn, Fe, Cu, and Mn in comparison with J774 and HeLa cells, indicating a neutrophil-specific metallome complying with their designated functions. When investigating PMA-activated neutrophils, the SR-XRF analysis depicted typical subcellular morphological changes: the transformation of nucleus and granules and the emergence of void vacuoles. Mature NETs were evenly composed of Fe, P, S, and Ca with occasional hot spots containing Zn, Fe, and Ca. An ICP-MS-based quantification of NET supernatants revealed a NETosis-induced decrease of soluble Zn, whereas Fe, Cu, and Mn concentrations were only slightly affected. In summary, we present a combination of SR-XRF and ICP-MS as a powerful tool to analyze trace elements in human neutrophils. The approach will be applicable and valuable to numerous aspects of nutritional immunity.

Graphical abstract: Trace element landscape of resting and activated human neutrophils on the sub-micrometer level

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Publication details

The article was received on 22 Dec 2014, accepted on 20 Mar 2015 and first published on 20 Mar 2015


Article type: Paper
DOI: 10.1039/C4MT00346B
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Citation: Metallomics, 2015,7, 996-1010
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    Trace element landscape of resting and activated human neutrophils on the sub-micrometer level

    M. J. Niemiec, B. De Samber, J. Garrevoet, E. Vergucht, B. Vekemans, R. De Rycke, E. Björn, L. Sandblad, G. Wellenreuther, G. Falkenberg, P. Cloetens, L. Vincze and C. F. Urban, Metallomics, 2015, 7, 996
    DOI: 10.1039/C4MT00346B

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