Influence of Age on the Release of Reactive Oxygen Species by Phagocytes as Measured by a Whole Blood Chemiluminescence Assay

Kukovetz, E.; Bratschitsch, G.; Hofer, H. P.; Egger, G.; Schaur, R. J.

doi:10.1016/s0891-5849(96)00334-6

Cited by 50 publications

(47 citation statements)

References 24 publications

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“…In addition, they did not find any inhibitory effects of phenolics at lower concentrations. The differences in our study may result from the use of a more sensitive and stable chemiluminometer together with the measurement of the oxidative response of PMNs in the diluted whole blood system, which avoids an unspecific activation of PMNs during the isolation procedure (Kukovetz et al, 1997). …”

Section: Relevance Of In Vitro Results To In Vivo Conditionsmentioning

confidence: 95%

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Inhibitory effect of plant phenolics on fMLP-induced intracellular calcium rise and chemiluminescence of human polymorphonuclear leukocytes and their chemotactic activityin vitro

Nowak

Zasowska-Nowak

Białasiewicz

et al. 2015

Pharmaceutical Biology

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Context: Polymorphonuclear leukocytes (PMNs) produce oxidants, contributing to systemic oxidative stress. Diets rich in plant polyphenols seem to decrease the risk of oxidative stressinduced disorders including cardiovascular disease. Objective: The objective of this study was to examine the in vitro effect of each of the 14 polyphenols on PMNs chemotaxis, intracellular calcium response, oxidants production. Materials and methods: Blood samples and PMNs suspensions were obtained from 60 healthy non-smoking donors and incubated with a selected polyphenol (0.5-10 mM) or a control solvent. We assessed resting and fMLP-dependent changes of intracellular calcium concentration ([Ca 2+ ] i ) in PMNs with the Fura-2AM method and measured fMLP-induced luminol enhanced whole blood chemiluminescence (fMLP-LBCL). Polyphenol chemoattractant activity for PMNs was tested with Boyden chambers. Results: Polyphenols had no effect on resting [Ca 2+ ] i . Unaffected by other compounds, fMLPdependent increase of [Ca 2+ ] i was inhibited by quercetin and catechol (5 mM) by 32 ± 14 and 12 ± 10% (p50.04), respectively. Seven of the 14 tested substances (5 mM) influenced fMLP-LBCL by decreasing it. Catechol, quercetin, and gallic acid acted most potently reducing fMLP-LBCL by 49 ± 5, 42 ± 15, and 28 ± 18% (p50.05), respectively. 3,4-Dihydroxyhydrocinnamic, 3,4-dihydroxyphenylacetic, 4-hydroxybenzoic acid, and catechin (5 mM) revealed distinct (p50.02) chemoattractant activity with a chemotactic index of 1.9 ± 0.8, 1.8 ± 0.7, 1.6 ± 0.6, 1.4 ± 0.2, respectively. Conclusion and discussion: Catechol, quercetin, and gallic acid at concentrations commensurate in human plasma strongly suppressed the oxidative response of PMNs. Regarding quercetin and catechol, this could result from an inhibition of [Ca 2+ ] i response.

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Section: Relevance Of In Vitro Results To In Vivo Conditionsmentioning

confidence: 95%

“…The effect of polyphenols on fMLP-stimulated luminalenhanced whole blood chemiluminescence (fMLP-LBCL) was measured according to Kukovetz et al (1997) with some modifications. Blood samples were prediluted with a mixture solution (30 mL of blood mixed with 1 mL of a mixture solution).…”

Section: Whole Blood Chemiluminescence Assaymentioning

confidence: 99%

Inhibitory effect of plant phenolics on fMLP-induced intracellular calcium rise and chemiluminescence of human polymorphonuclear leukocytes and their chemotactic activityin vitro

Nowak

Zasowska-Nowak

Białasiewicz

et al. 2015

Pharmaceutical Biology

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“…The lucigenin-perborate ROS-generating system used in our study gives a strong and constant light emission for more than 15 min, thus avoiding false results associated with a decreasing source intensity. The AOA test is simple, fast and inexpensive when compared to other multi-step CL techniques [11,12,21]. It also enables the AOA evaluation of different drug and dietary agents and their effects on the total antioxidant status of the body [26].…”

Section: Discussionmentioning

confidence: 99%

“…However, different methods used for the purification of phagocytes from the whole blood may change the initial activation state of these cells by depleting the environmental cytokines, complement factors, bacterial products and immune complexes present in plasma and whole blood. Alternatively, they may reduce the numbers of primed phagocytes through clumping and adhesion to the glass walls or may record a different activation state after time-consuming purification procedures (1±2 h) [21]. Direct functional analysis of the circulating phagocyte activity in whole blood is therefore thought to minimize artifacts introduced by cell purification, reduce the time required for each test, and maintain the in vivo cellular environment.…”

Section: Basal Photon Emission In Venous Bloodmentioning

confidence: 99%

“…Simple Chemiluminescence Assays for Free Radicals in Venous Blood and Serum Samples Instead of using polystyrene beads [21] or phorbol 12-myristate 13-acetate [22] for maximal ROS production, our test induced a similar effect after 3 min of light exposure without any additional stimulatory compound. Treatment of light-exposed whole-blood samples with SOD (1,250 IU) resulted in an 85% inhibition of the light output, strongly suggesting an involvement of the superoxide anion in this process.…”

Section: Light Exposure Test In Venous Bloodmentioning

confidence: 99%

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Simple Chemiluminescence Assays for Free Radicals in Venous Blood and Serum Samples: Results in Atopic, Psoriasis, MCS and Cancer Patients

Ionescu¹,

Merk²,

Bradford³

1999

Complement Med Res

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Objective: To investigate the generation of reactive oxygen species (ROS) in serum and venous blood as well as the serum antioxidative activity (AOA) in patients and healthy controls by means of a simplified chemiluminescence (CL) methodology. Study Participants:48 Atopic eczema, 23 psoriasis, 15 multiple chemical sensitivity (MCS) and 35 cancer patients together with 22 healthy volunteers. Methods:ROS generation/photon emission in blood and serum samples under basal conditions and after light exposure as well as the AOA of the serum samples was investigated at room temperature (22 °C) in all 143 fasted subjects. The 3-step methodology resumes in adding a constant amount of blood or serum to a constant amount of CL substrate (or to an ROS-generating mixture for the AOA test), followed by a short preincubation and registration of the photon counts over a 600-second time interval. Results:In the basal and light exposure tests significantly higher photon counts (> 14,000 counts/600 s) were registered in venous blood in all patient groups when compared to healthy controls (p < 0.001), suggesting increased amounts of activated leukocytes and light-sensitizing compounds, respectively. By contrast, most patient sera showed in all three CL tests a strongly inhibited light emission (p < 0.005), suggesting an adaptive antioxidative response to oxidant stress factors. Conclusions:Atopic, psoriasis, MCS and cancer patients are exhibiting significantly changed blood and serum CL patterns when compared to healthy controls. The described assays are simple, well reproducible and enable a fast assessment of ROS generation and AOA in biological samples at low operational costs.

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The phagocyte chemiluminescence paradox: luminol can act as an inhibitor of neutrophil NADPH-oxidase activity

et al. 1999

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The chemiluminescence system amplified by luminol or isoluminol is a sensitive and widely used method for determination of respiratory burst products generated by the NADPH‐oxidase in phagocytes. The present study shows that luminol, but not isoluminol, can inhibit the release of oxygen metabolites generated by human neutrophil NADPH‐oxidase. The difference in structure between luminol and isoluminol (rendering luminol more lipophilic than isoluminol, and thereby membrane‐permeable), is suggested to determine indirectly whether or not the molecule is inhibitory. Luminol was shown to have an increased inhibitory effect after preincubation of neutrophils on a surface of aggregated IgG, suggesting that the cells can be transferred from a ‘luminol‐insensitive’ to a ‘luminol‐sensitive’ state. Since luminol had no inhibitory effect in a cell‐free NADPH‐oxidase system, it is likely that it interferes with the signal transduction pathway, leading to assembly and/or activation of the oxidase. As a consequence of the present results, showing that luminol but not isoluminol can inhibit NADPH‐oxidase activity, we suggest that isoluminol is used in future studies of superoxide anion release from phagocytes. Copyright © 1999 John Wiley & Sons, Ltd.

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Influence of Age on the Release of Reactive Oxygen Species by Phagocytes as Measured by a Whole Blood Chemiluminescence Assay

Cited by 50 publications

References 24 publications

Inhibitory effect of plant phenolics on fMLP-induced intracellular calcium rise and chemiluminescence of human polymorphonuclear leukocytes and their chemotactic activityin vitro

Inhibitory effect of plant phenolics on fMLP-induced intracellular calcium rise and chemiluminescence of human polymorphonuclear leukocytes and their chemotactic activityin vitro

Simple Chemiluminescence Assays for Free Radicals in Venous Blood and Serum Samples: Results in Atopic, Psoriasis, MCS and Cancer Patients

The phagocyte chemiluminescence paradox: luminol can act as an inhibitor of neutrophil NADPH-oxidase activity

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