Evaluation of whole antioxidant defenses of human mononuclear cells by a new in vitro biological test: lack of correlation between erythrocyte and mononuclear cell resistance to oxidative stress

Sylvie Caspar-Bauguil , Nicolas Maestre , Céline Segafredo , Anne Galinier , Jésus Garcia , Michel Prost , Brigitte Périquet , Luc Pénicaud (1) , Robert Salvayre , Louis Casteilla

1 DCCI - Développement et Communication Chimique chez les Insectes

OBJECTIVES: This work aims to evaluate the resistance of mononuclear cells to oxidative stress using a "KRL" test, formerly utilized to evaluate the resistance of erythrocyte to free radicals. METHODS: The "KRL" test evaluates the resistance to lysis of cells treated by free radicals generated under standardized conditions. RESULTS: We defined new analytical parameters (level of radical production, time course, number of cells) to obtain an accurate assay determining the resistance to oxidative stress of mononuclear cells, in comparison to that of erythrocytes. This test allows the evaluation of change in the redox state of mononuclear cells (improved by an antioxidant mix or deteriorated by antimycin A-induced mitochondrial radical overproduction). Interestingly, our data show that the sensitivity of mononuclear cells to oxidative stress is not correlated with the susceptibility of erythrocytes to oxidative stress. CONCLUSIONS: The quantification of the susceptibility of mononuclear cells to oxidative stress gives additional information (in addition to erythrocyte resistance) and could be helpful for patients with chronic inflammation.

Mots clés: Anti-Bacterial Agents/pharmacology; Antimycin A/pharmacology; Antioxidants/*metabolism/pharmacology; Cell Culture Techniques; Electron Transport Complex III/antagonists & inhibitors; Erythrocytes/*metabolism; Humans; Immunologic Tests/*methods; Leukocytes; Mononuclear/drug effects/*metabolism; Oxidation-Reduction; Oxidative Stress/*drug effects; Reactive Oxygen Species/metabolism; Reagent Kits; Diagnostic; Reproducibility of Results; Statistics as Topic

Sylvie Caspar-Bauguil, Nicolas Maestre, Céline Segafredo, Anne Galinier, Jésus Garcia, et al.. Evaluation of whole antioxidant defenses of human mononuclear cells by a new in vitro biological test: lack of correlation between erythrocyte and mononuclear cell resistance to oxidative stress. Clin Biochem, 2009, 42 (6), pp.510-4. ⟨hal-00461350⟩