Quantitative Magnetic Resonance Imaging of Enzyme Activity on the Cell Surface: In Vitro and In Vivo Monitoring of ADP-Ribosyltransferase 2 on T Cells
The objective of this study was to quantify enzymatic activity on the surface of T cells by magnetic resonance imaging (MRI) using R2 and R2* relaxometry. Lymphoma cells expressing adenosine diphosphate (ADP)-ribosyltransferase 2 (ART2) were incubated with increasing doses of its substrate etheno–ni...
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2010-07-01
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| Series: | Molecular Imaging |
| Online Access: | https://doi.org/10.2310/7290.2010.00017 |
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| author | Peter Bannas Oliver Graumann Philipp Balcerak Kersten Peldschus Michael Gerhard Kaul Heinrich Hohenberg Friedrich Haag Gerhard Adam Harald Ittrich Friedrich Koch-Nolte |
| author_facet | Peter Bannas Oliver Graumann Philipp Balcerak Kersten Peldschus Michael Gerhard Kaul Heinrich Hohenberg Friedrich Haag Gerhard Adam Harald Ittrich Friedrich Koch-Nolte |
| author_sort | Peter Bannas |
| collection | DOAJ |
| description | The objective of this study was to quantify enzymatic activity on the surface of T cells by magnetic resonance imaging (MRI) using R2 and R2* relaxometry. Lymphoma cells expressing adenosine diphosphate (ADP)-ribosyltransferase 2 (ART2) were incubated with increasing doses of its substrate etheno–nicotinamide adenine dinucleotide (NAD), resulting in increasing amounts of surface protein ADP-ribosylation. Etheno-ADP-ribosylated proteins were detected with monoclonal antibody 1G4 and superparamagnetic iron oxide conjugated secondary antibodies (Ab-SPIO). Labeling efficiency was determined with R2 and R2* relaxometry on a clinical 3.0 T scanner. Parallel aliquots of cells were analyzed by flow cytometry. Cell-bound SPIO conjugates were detected by immunofluorescence and electron microscopy and quantified by atomic absorption spectroscopy. To mimic an inflammatory site in vivo, Ab-SPIO-labeled cells were injected subcutaneously in mice and analyzed by MRI. Immunofluorescence and electron microscopy confirmed cell-surface localization of Ab-SPIO. MRI of Ab-SPIO-labeled cells showed a corresponding signal reduction. Increases in R2 and R2* determined by magnetic resonance relaxometry correlated linearly with the expression level of ART2 and the concentration of the ART2 substrate etheno-NAD. R2 and R2* increases correlated linearly with the results from flow cytometry and atomic absorption spectroscopy analyses. Quantitative R2 and R2* mapping enable noninvasive determination of enzymatic activity on T cells and holds promise for characterization of inflammatory sites in vivo by MRI. |
| format | Article |
| id | doaj-art-7bd6d31f9a43469c9d6ce7a96544eaf3 |
| institution | Kabale University |
| issn | 1536-0121 |
| language | English |
| publishDate | 2010-07-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Molecular Imaging |
| spelling | doaj-art-7bd6d31f9a43469c9d6ce7a96544eaf32025-01-03T00:10:30ZengSAGE PublishingMolecular Imaging1536-01212010-07-01910.2310/7290.2010.0001710.2310_7290.2010.00017Quantitative Magnetic Resonance Imaging of Enzyme Activity on the Cell Surface: In Vitro and In Vivo Monitoring of ADP-Ribosyltransferase 2 on T CellsPeter BannasOliver GraumannPhilipp BalcerakKersten PeldschusMichael Gerhard KaulHeinrich HohenbergFriedrich HaagGerhard AdamHarald IttrichFriedrich Koch-NolteThe objective of this study was to quantify enzymatic activity on the surface of T cells by magnetic resonance imaging (MRI) using R2 and R2* relaxometry. Lymphoma cells expressing adenosine diphosphate (ADP)-ribosyltransferase 2 (ART2) were incubated with increasing doses of its substrate etheno–nicotinamide adenine dinucleotide (NAD), resulting in increasing amounts of surface protein ADP-ribosylation. Etheno-ADP-ribosylated proteins were detected with monoclonal antibody 1G4 and superparamagnetic iron oxide conjugated secondary antibodies (Ab-SPIO). Labeling efficiency was determined with R2 and R2* relaxometry on a clinical 3.0 T scanner. Parallel aliquots of cells were analyzed by flow cytometry. Cell-bound SPIO conjugates were detected by immunofluorescence and electron microscopy and quantified by atomic absorption spectroscopy. To mimic an inflammatory site in vivo, Ab-SPIO-labeled cells were injected subcutaneously in mice and analyzed by MRI. Immunofluorescence and electron microscopy confirmed cell-surface localization of Ab-SPIO. MRI of Ab-SPIO-labeled cells showed a corresponding signal reduction. Increases in R2 and R2* determined by magnetic resonance relaxometry correlated linearly with the expression level of ART2 and the concentration of the ART2 substrate etheno-NAD. R2 and R2* increases correlated linearly with the results from flow cytometry and atomic absorption spectroscopy analyses. Quantitative R2 and R2* mapping enable noninvasive determination of enzymatic activity on T cells and holds promise for characterization of inflammatory sites in vivo by MRI.https://doi.org/10.2310/7290.2010.00017 |
| spellingShingle | Peter Bannas Oliver Graumann Philipp Balcerak Kersten Peldschus Michael Gerhard Kaul Heinrich Hohenberg Friedrich Haag Gerhard Adam Harald Ittrich Friedrich Koch-Nolte Quantitative Magnetic Resonance Imaging of Enzyme Activity on the Cell Surface: In Vitro and In Vivo Monitoring of ADP-Ribosyltransferase 2 on T Cells Molecular Imaging |
| title | Quantitative Magnetic Resonance Imaging of Enzyme Activity on the Cell Surface: In Vitro and In Vivo Monitoring of ADP-Ribosyltransferase 2 on T Cells |
| title_full | Quantitative Magnetic Resonance Imaging of Enzyme Activity on the Cell Surface: In Vitro and In Vivo Monitoring of ADP-Ribosyltransferase 2 on T Cells |
| title_fullStr | Quantitative Magnetic Resonance Imaging of Enzyme Activity on the Cell Surface: In Vitro and In Vivo Monitoring of ADP-Ribosyltransferase 2 on T Cells |
| title_full_unstemmed | Quantitative Magnetic Resonance Imaging of Enzyme Activity on the Cell Surface: In Vitro and In Vivo Monitoring of ADP-Ribosyltransferase 2 on T Cells |
| title_short | Quantitative Magnetic Resonance Imaging of Enzyme Activity on the Cell Surface: In Vitro and In Vivo Monitoring of ADP-Ribosyltransferase 2 on T Cells |
| title_sort | quantitative magnetic resonance imaging of enzyme activity on the cell surface in vitro and in vivo monitoring of adp ribosyltransferase 2 on t cells |
| url | https://doi.org/10.2310/7290.2010.00017 |
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