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Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry

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Al-Sandaqchi, Alaa T., Brignell, Chris, Collingwood, Joanna F., Geraki, Kalotina, Mirkes, Evgeny M., Kong, Kenny, Castellanos, Marcos, May, Sean T., Stevenson, Carl W. and Elsheikha, Hany M. (2018) Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry. Metallomics, 10 (10). pp. 1401-1414. doi:10.1039/c8mt00136g ISSN 1756-5901.

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Official URL: https://doi.org/10.1039/C8MT00136G

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Abstract

In this study, we measured the levels of elements in human brain microvascular endothelial cells (ECs) infected with T. gondii. ECs were infected with tachyzoites of the RH strain, and at 6, 24, and 48 hours post infection (hpi), the intracellular concentrations of elements were determined using a synchrotron-microfocus X-ray fluorescence microscopy (μ-XRF) system. This method enabled the quantification of the concentrations of Zn and Ca in infected and uninfected (control) ECs at sub-micron spatial resolution. T. gondii-hosting ECs contained less Zn than uninfected cells only at 48 hpi (p < 0.01). The level of Ca was not significantly different between infected and control cells (p > 0.05). Inductively Coupled Plasma Mass Spectrometry (ICP-MS) analysis revealed infection-specific metallome profiles characterized by significant increases in the intracellular levels of Zn, Fe, Mn and Cu at 48 hpi (p < 0.01), and significant reductions in the extracellular concentrations of Co, Cu, Mo, V, and Ag at 24 hpi (p < 0.05) compared with control cells. Zn constituted the largest part (74%) of the total metal composition (metallome) of the parasite. Gene expression analysis showed infection-specific upregulation in the expression of five genes, MT1JP, MT1M, MT1E, MT1F, and MT1X, belonging to the metallothionein gene family. These results point to a possible correlation between T. gondii infection and increased expression of MT1 isoforms and altered intracellular levels of elements, especially Zn and Fe. Taken together, a combined μ-XRF and ICP-MS approach is promising for studies of the role of elements in mediating host-parasite interaction.

Item Type: Journal Article
Subjects: Q Science > QL Zoology
Q Science > QP Physiology
Divisions: Faculty of Science, Engineering and Medicine > Engineering > Engineering
SWORD Depositor: Library Publications Router
Library of Congress Subject Headings (LCSH): Endothelial cells, Toxoplasma gondii -- Infections
Journal or Publication Title: Metallomics
Publisher: Royal Society of Chemistry
ISSN: 1756-5901
Official Date: 17 October 2018
Dates:
DateEvent
17 October 2018Published
5 September 2018Available
14 August 2018Accepted
Volume: 10
Number: 10
Page Range: pp. 1401-1414
DOI: 10.1039/c8mt00136g
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Open Access (Creative Commons)
Date of first compliant deposit: 10 October 2018
Date of first compliant Open Access: 5 September 2019
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
UNSPECIFIED[STFC] Science and Technology Facilities Councilhttp://dx.doi.org/10.13039/501100000271
SP8504Diamond Light Sourcehttp://dx.doi.org/10.13039/100011889
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