Fig. 4

Immunosuppressive and neuroprotective effects of cladribine are transient on cellular level. A Oral treatment with cladribine induced a less severe disease score on day 27 in EAE mice compared to EAE mice receiving vehicle treatment (unpaired Mann–Whitney U test, p = 0.0386, cladribine-treated (n = 12) vs. vehicle-treated (n = 13)). B Immunophenotypings of the peripheral blood (a), LNs (b), spleen (c), thymus (d) and bone marrow (e) at day 27 post-EAE induction were performed by flow cytometry. Immune cell profiles of mice treated with cladribine were compared to those receiving vehicle. No statistically significant differences could be observed (two-way ANOVAs, n = 3 for each group. p-values > 0.05). C: Flow cytometric analysis of the immune cell distribution in the brain (a) and spinal cord (b) of EAE mice on day 27 post-immunization, either treated with cladribine or vehicle, shows no significant differences between both experimental groups (two-way ANOVAs, n = 3 for each group. p-values > 0.05). D mRNA expression of deoxycytidine kinase (DCK) was quantified by quantitative polymerase chain reaction (qPCR) in murine PBMCs and normalized to human PBMCs. Results are depicted as 2^−∆∆Ct values (n = 6 for both species, unpaired Mann–Whitney U test, p = 0.0022). E Electrophysiological recordings of APs were obtained by recording in current-clamp mode. Mean bar graph indicating the number of APs recorded on day 27 post-EAE induction in response to depolarizing steps of increased intensity ranging from + 20 to + 160 pA. No significant alterations could be observed between groups (two-way ANOVA, n = 23—naïve ctrl, 10—vehicle, 9—cladribine. p-values > 0.05). p > 0.05 = ns, p < 0.05 = *, p < 0.01 = **