Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry
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 s...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Royal Society of Chemistry
2018
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| Online Access: | https://eprints.nottingham.ac.uk/55172/ |
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| author | Al-sandaqchi, Alaa T. Brignell, Chris Collingwood, Joanna F. Geraki, Kalotina Mirkes, Evgeny M. Kong, Kenny Castellanos, Marcos May, Sean T. Stevenson, Carl W. Elsheikha, Hany M. |
| author_facet | Al-sandaqchi, Alaa T. Brignell, Chris Collingwood, Joanna F. Geraki, Kalotina Mirkes, Evgeny M. Kong, Kenny Castellanos, Marcos May, Sean T. Stevenson, Carl W. Elsheikha, Hany M. |
| author_sort | Al-sandaqchi, Alaa T. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T20:30:46Z |
| format | Article |
| id | nottingham-55172 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T20:30:46Z |
| publishDate | 2018 |
| publisher | Royal Society of Chemistry |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-551722019-09-05T04:30:53Z https://eprints.nottingham.ac.uk/55172/ Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry Al-sandaqchi, Alaa T. Brignell, Chris Collingwood, Joanna F. Geraki, Kalotina Mirkes, Evgeny M. Kong, Kenny Castellanos, Marcos May, Sean T. Stevenson, Carl W. Elsheikha, Hany M. 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. Royal Society of Chemistry 2018-09-05 Article PeerReviewed application/pdf en https://eprints.nottingham.ac.uk/55172/1/Author%20accepted%20manuscript_Alsandaqchi%20et%20al-Metallomics.pdf 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 . ISSN 1756-591X Toxoplasma gondii blood-brain barrier host-pathogen interaction label-free imaging X-ray fluorescence spectromicroscopy; Mass Spectrometry http://pubs.rsc.org/en/Content/ArticleLanding/2018/MT/C8MT00136G#!divAbstract doi:10.1039/C8MT00136G doi:10.1039/C8MT00136G |
| spellingShingle | Toxoplasma gondii blood-brain barrier host-pathogen interaction label-free imaging X-ray fluorescence spectromicroscopy; Mass Spectrometry Al-sandaqchi, Alaa T. Brignell, Chris Collingwood, Joanna F. Geraki, Kalotina Mirkes, Evgeny M. Kong, Kenny Castellanos, Marcos May, Sean T. Stevenson, Carl W. Elsheikha, Hany M. Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry |
| title | Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry |
| title_full | Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry |
| title_fullStr | Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry |
| title_full_unstemmed | Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry |
| title_short | Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry |
| title_sort | metallome of cerebrovascular endothelial cells infected with toxoplasma gondii using μ-xrf imaging and inductively coupled plasma mass spectrometry |
| topic | Toxoplasma gondii blood-brain barrier host-pathogen interaction label-free imaging X-ray fluorescence spectromicroscopy; Mass Spectrometry |
| url | https://eprints.nottingham.ac.uk/55172/ https://eprints.nottingham.ac.uk/55172/ https://eprints.nottingham.ac.uk/55172/ |