Intracellular oxidative activity and respiratory burst of leukocytes
isolated from multiple sclerosis patients.
Ferretti G, Bacchetti T, Diludovico F, Viti B, Angeleri VA, Danni M,
Provinciali L
Neurochem Int. 2005 Oct 28;

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(2)DCFDA) 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.

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