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Evaluation of electron spin resonance for studies of superoxide anion production by human neutrophils interacting with Staphylococcus aureus and Staphylococcus epidermidis.
Department of Molecular and Clinical Medicine Linköping University.
Department of Neuroscience and Locomotion Linköping University.
Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för klinisk och experimentell medicin, Experimentell patologi.
Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för medicin och hälsa, Medicinsk radiofysik.
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2008 (engelsk)Inngår i: Journal of Biochemical and Biophysical Methods, ISSN 0165-022X, E-ISSN 1872-857X, Vol. 70, nr 6, s. 1059-1065Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

The present study evaluates electron spin resonance (ESR) and the spin trapper 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide (DEPMPO) for analysis of superoxide radical production by human neutrophils interacting with viable Staphylococcus aureus and Staphylococcus epidermidis bacteria. To avoid auto-activation due to interaction with glass surfaces, neutrophils were preincubated in plastic tubes until the peak response was reached, and then transferred to a quartz flat cell to record the ESR spectra. The time point for peak response was identified by parallel analysis of the bacteria-neutrophil interaction using luminol amplified chemiluminescence. We found detectable ESR spectra from neutrophils interacting with as few as five bacteria of the weak activating S. epidermidis per neutrophil. Addition of the NADPH oxidase inhibitor diphenylene iodonium totally abolished spectra. Catalase, DMSO or an iron chelator had no impact on the produced spectra and ionomycin, a selective activator of intracellular NADPH oxidase, gave significant ESR spectra. Taken together, our results indicate that DEPMPO is cell permeable and detects NADPH oxidase derived superoxide anions formed in phagosomes or released by human neutrophils phagocytosing viable S. aureus and S. epidermidis. The technique may be used as a sensitive tool to evaluate superoxide anion production in human neutrophils.

sted, utgiver, år, opplag, sider
2008. Vol. 70, nr 6, s. 1059-1065
Emneord [en]
Cells, Cultured Electron Spin Resonance Spectroscopy/*methods Humans Neutrophils/chemistry/*metabolism Staphylococcus aureus/*physiology Staphylococcus epidermidis/*physiology Subcellular Fractions Superoxides/*analysis/chemistry/*metabolism
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Identifikatorer
URN: urn:nbn:se:liu:diva-43449DOI: 10.1016/j.jbbm.2007.05.014Lokal ID: 73882OAI: oai:DiVA.org:liu-43449DiVA, id: diva2:264308
Tilgjengelig fra: 2009-10-10 Laget: 2009-10-10 Sist oppdatert: 2017-12-13

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Johansson, UnoGustafsson, HåkanÖllinger, Karin

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