使用 μ-XRF 成像和电感耦合等离子体质谱法检测感染刚地弓形虫的脑血管内皮细胞的金属组†,Metallomics

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使用 μ-XRF 成像和电感耦合等离子体质谱法检测感染刚地弓形虫的脑血管内皮细胞的金属组†
Metallomics ( IF 2.9 ) Pub Date : 2018-09-05 00:00:00 , DOI: 10.1039/c8mt00136g
Alaa T Al-Sandaqchi ₁ , Chris Brignell ₂ , Joanna F Collingwood ₃ , Kalotina Geraki ₄ , Evgeny M Mirkes ₅ , Kenny Kong ₆ , Marcos Castellanos ₇ , Sean T May ₇ , Carl W Stevenson ₁ , Hany M Elsheikha ₈

Affiliation

在这项研究中，我们测量了感染弓形虫的人脑微血管内皮细胞 (EC) 中元素的水平。用 RH 菌株的速殖子感染 EC，在感染后 6、24 和 48 小时 (hpi)，使用同步加速器-微焦点 X 射线荧光显微镜 (μ-XRF) 系统测定细胞内元素的浓度。该方法能够以亚微米空间分辨率量化感染和未感染（对照）EC 中 Zn 和 Ca 的浓度。仅在 48 hpi ( p < 0.01)时， T. gondii宿主 EC 的 Zn 含量低于未感染细胞。Ca 水平在感染细胞和对照细胞之间没有显着差异（p> 0.05)。电感耦合等离子体质谱 (ICP-MS) 分析揭示了感染特异性金属组谱，其特征是 48 hpi 时细胞内 Zn、Fe、Mn 和 Cu 水平显着增加 ( p < 0.01)，而细胞外浓度显着降低与对照细胞相比， 24 hpi ( p < 0.05)时的 Co、Cu、Mo、V 和 Ag 。Zn 占寄生虫总金属成分（金属组）的最大部分（74%）。基因表达分析显示，属于金属硫蛋白基因家族的五个基因 MT1JP、MT1M、MT1E、MT1F 和 MT1X 的表达存在感染特异性上调。这些结果表明T. gondii之间可能存在相关性感染和 MT1 亚型的表达增加以及细胞内元素水平的改变，尤其是 Zn 和 Fe。总之，μ-XRF 和 ICP-MS 相结合的方法有望用于研究元素在介导宿主-寄生虫相互作用中的作用。

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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 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.

更新日期：2018-09-05

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