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Apoptotic markers in the peripheral blood and cerebrospinal fluid in patients with multiple sclerosis

Recent data indicate that the apoptotic process, mediated by the CD95/Fas cell surface receptor, is impaired in activated lymphocytes of patients with relapsing-remitting multiple sclerosis (RRMS). However, the role of lymphocyte apoptosis in patients with primary progressive multiple sclerosis (PPMS) is not completely explained in the literature. We presented results of the study performed in the Department of Neurology and Division of Immunology (Department of Clinical Laboratory Diagnostics) of the Zagreb University Hospital Centre. The study included 50 patients with multiple sclerosis diagnosed by McDonald’ s criteria (27 patients with RRMS, 10 patients with PPMS, 13 patients with secondary progressive multiple sclerosis - SPMS) and 31 controls. Apoptosis of peripheral blood (PB) lymphocytes was analysed by the use of annexin-V and propidium iodide labelling as well as flow cytometry. The proportion of PB CD4+, CD8+, CD4+CD95+ and CD8+CD95+ T lymphocytes was analysed by flow cytometric-immunophenotyping. The stated parameters were determined before and after pulse corticosteroid therapy, as well as six months after the beginning of investigation (patients with RRMS were then in the remission). Apoptosis of cerebrospinal fluid (CSF) lymphocytes was analysed by the use of acridin-orange-ethidium-bromide labelling and was done in all patients once, before therapy. In all patients significant decrease in the proportion of CD4+CD95+ and CD8+CD95+ T lymphocytes and increase in the lymphocyte apoptosis in blood was found after therapy, as well as significant difference towards controls. However, six months after beginning of investigation RRMS patients had significantly lower proportion of CD4+CD95+ and CD8+CD95+ T lymphocytes and higher proportion of apoptotic lymphocytes in comparison to the values at the beginning of investigation, while in patients with progressive form of the disease the values of investigated parameters were similar to those at the beginning of investigation, before therapy. The proportion of apoptotic lymphocytes in CSF was significantly lower in all three groups of patients compared to controls, and also significantly lower in patients with RRMS compared to patients with PPMS and SPMS. The differences in dynamics of stated laboratory parameters, indicators of blood and CSF lymphocyte apoptosis, point to the different patophysiologic events in patients with RRMS compared to the patients with PPMS and SPMS.

apoptosis; CD95/Fas; lymphocytes; relapsing-remitting multiple sclerosis; primary progressive multiple sclerosis

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