Figure 5 Treatment with fingolimod raises the activation threshold of monocytes in MS Peripheral blood mononuclear cells from 8 healthy donors, 7 patients.

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Figure 5 Treatment with fingolimod raises the activation threshold of monocytes in MS Peripheral blood mononuclear cells from 8 healthy donors, 7 patients with untreated multiple sclerosis (MS), and 11 patients with fingolimod-treated MS were briefly stimulated ex vivo with lipopolysaccharide (LPS). Treatment with fingolimod raises the activation threshold of monocytes in MS Peripheral blood mononuclear cells from 8 healthy donors, 7 patients with untreated multiple sclerosis (MS), and 11 patients with fingolimod-treated MS were briefly stimulated ex vivo with lipopolysaccharide (LPS). Monocytes were gated by forward vs side scatterplot and CD14-positive staining. Gate on viable (7-AAD negative) cells was then applied. Monocyte activation was monitored by staining for the 2 surface markers CD25 and CD150. Thresholds were set on relative isotype controls. (A) Frequency of CD25-positive (upper graph) and CD150-positive (lower graph) monocytes cultured ex vivo with 100 ng/mL LPS; each dot represents a single participant; black bars represent mean values for each group. (B) CD25 (upper) and CD150 (lower) stainings in cultures stimulated with 600 pg/mL LPS from a representative healthy control (left), a patient with untreated MS (middle), and a patient with fingolimod-treated MS (right). (C) Frequency of CD25 and CD150 positive monocytes at increasing LPS concentrations. Dose response curves are shown for each participant. Right panels show the means for each group. *p < 0.05; ***p < 0.001. FSC = forward scatter. Marco Di Dario et al. Neurol Neuroimmunol Neuroinflamm 2015;2:e157 © 2015 American Academy of Neurology