Intracellular oxidative activity and respiratory burst of leukocytes isolated from multiple sclerosis patients

Oxidative damage induced by free radicals and reactive oxygen species (ROS) have been suggested to play an important role in the development of autoimmune diseases such as multiple sclerosis (MS) disease and it has been hypothesised that oxidative injury could mediate demyelination and axonal injury...

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Published in:	Neurochemistry international Vol. 48; no. 2; pp. 87 - 92
Main Authors:	Ferretti, G., Bacchetti, T., DiLudovico, F., Viti, B., Angeleri, V.A., Danni, M., Provinciali, L.
Format:	Journal Article
Language:	English
Published:	Oxford Elsevier Ltd 2006 Elsevier
Subjects:	2′-7′-dichlorodihydrofluorescin diacetate (H 2DCFDA) Adult Biological and medical sciences Case-Control Studies Evoked Potentials, Visual Female Fluoresceins - chemistry Fluorescent Dyes - chemistry Fundamental and applied biological sciences. Psychology Humans Leukocytes Leukocytes - drug effects Leukocytes - metabolism Magnetic Resonance Imaging Male Medical sciences Middle Aged Multiple sclerosis Multiple Sclerosis - blood Multiple Sclerosis - metabolism Multiple Sclerosis - physiopathology Multiple sclerosis and variants. Guillain barré syndrome and other inflammatory polyneuropathies. Leukoencephalitis Neurology Oxidative damage Oxidative Stress Respiratory Burst Tetradecanoylphorbol Acetate - pharmacology Vertebrates: nervous system and sense organs 2′-7′-dichlorodihydrofluorescin diacetate (H 2DCFDA) Multiple sclerosis Leukocytes Respiratory burst Oxidative damage Human Nervous system diseases Leukocyte Central nervous system disease Intracellular 2'-7'-dichlorodihydrofluorescin diacetate (H2DCFDA) Inflammatory disease
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Summary:	Oxidative damage induced by free radicals and reactive oxygen species (ROS) have been suggested to play an important role in the development of autoimmune diseases such as multiple sclerosis (MS) disease and it has been hypothesised that oxidative injury could mediate demyelination and axonal injury in MS subjects. In our study, we compared intracellular oxidative activity and the respiratory burst activity in MS patients ( n = 20) and healthy controls ( n = 15) using leukocytes as cellular model. At this purpose, intracellular ROS levels were evaluated by fluorometric assay using the 2′-7′-dichlorodihydrofluorescin diacetate probe (H 2DCFDA) in untreated or in leukocytes stimulated with phorbol-12-myristate-13-acetate (PMA). Our results demonstrate that the intracellular spontaneous ROS production in leukocytes from MS patients was higher with respect to cells from control subjects ( p < 0.001). PMA addition induced a higher formation of ROS both in leukocytes from MS patients and controls ( p < 0.001). The PMA-induced production of ROS was significantly higher in leukocytes from MS with respect to controls ( p < 0.001). Significant positive correlations were established between intracellular spontaneous or PMA-induced production of ROS in leukocytes isolated from MS patients and the clinical parameters used to evaluate disease disability such as expanded disability status scale (EDSS), brain lesions evaluated by MRI and visual evoked potential (VEP) ( p < 0.001). In conclusion, our results demonstrate higher levels of intracellular ROS in untreated or in PMA-treated leukocytes isolated from MS patients with respect to healthy subjects confirming the role of oxidative stress in multiple sclerosis.
ISSN:	0197-0186 1872-9754
DOI:	10.1016/j.neuint.2005.09.005