Table 1.
Patient and healthy control (HC) characteristics at baseline.
Table 2.
Clinical parameters at baseline and at follow-up after 1-year fingolimod treatment.
Table 3.
Characteristics of patients (n = 10) showing disease activity during follow-up (EDA).
Fig 1.
Distribution of lymphocytes in fingolimod treated patients and healthy controls.
(a) Scatter plots showing the number (x106/L) of lymphocytes, T helper (Th) cells, cytotoxic T (Tc) cells, B cells, NK cells and the ratio of CD4+/CD8+ (mean and SD) at baseline and after one year of fingolimod treatment in MS patients (n = 19) and in healthy controls (n = 18). (b) Pie charts illustrating the change in relative mean distribution of Th, Tc, B and NK cells at baseline, at the 1-year-follow up and in healthy controls. (c) Scatter plots highlighting the previous disease modifying treatment and the number of lymphocytes and subpopulations at baseline and after treatment. Red = natalizumab, blue = interferon-β, black = untreated. (d) Scatter plots excluding previously natalizumab-treated patients at baseline and after one year of fingolimod therapy.
Fig 2.
Gating strategy of lymphocytes and T-cell subpopulations.
(a) Representative flow cytometric dot plots illustrating the gating strategy of lymphocytes and T cells (CD3+4+ and CD3+8+). (b) Representative T-cell differentiation pattern with respect to CD45RA and CD62L expression and the effect of fingolimod treatment in CD3+4+ (left column) and CD3+8+ cells (right column). The top panels indicate a typical distribution at baseline while the lower panels show the radical redistribution after one year of fingolimod treatment. Naïve cells were gated as positive for both CD45RA and the ligand for lymph node homing, CD62L; central memory (CM) as CD45RA-CD62L+; effector memory (EM) as CD45RA-CD62L- and terminally differentiated effector memory RA-positive cells (TEMRA) as CD45RA+CD62L-.
Fig 3.
Effect of fingolimod on differentiation pattern of CD4+ and CD8+ cells.
(a) Scatter plots showing the number (x106/L) of CD4+ and (b) CD8+ cells (mean and SD) from patients (n = 18) at baseline and after 1-year of fingolimod treatment. (c) Pie charts illustrating the change in relative mean distribution of CD4+ and (d) CD8+ cells at baseline and after 1 year. Subpopulations are defined as naïve, central memory (CM), effector memory (EM) and terminally differentiated effector memory cells (TEMRA) depending on their expression of CD45RA and CD62L.
Fig 4.
Effect of fingolimod on regulatory T cells (Tregs).
(a) The number (x106/L) of Tregs (sum of naive (FOXP3dimCD45RA+) and memory (FOXP3+CD45RA-)) at baseline and after 1-year fingolimod treatment (n = 19). (b) Both the naive and (c) memory Tregs were reduced in numbers. (d) The lower panel shows the percentage of Tregs at baseline and after one year; sum of naive and memory, (e) exclusively naive and (f) exclusively memory.
Fig 5.
Effect of fingolimod on activation markers.
(a) Scatter plots showing the percentage (mean and SD) of CD4+ cells expressing the activation markers CD69 and (b) HLA-DR in patients at baseline and after 1-year fingolimod treatment (n = 19). (c) CD69 and (d) HLA-DR expression in CD8+ cells and in (e-f) NK cells.
Fig 6.
Differences in baseline characteristics of NEDA compared to EDA patients.
(a) Scatter plots showing the baseline percentage of T helper and (b) NK cells expressing the activation marker HLA-DR (mean and SD) in patients with no evidence of disease activity (NEDA) and in patients with evidence of disease activity (EDA) (n = 19). (c) The number (x106/L) of terminally differentiated effector memory (TEMRA) T helper cells at baseline and after one year in NEDA and EDA patients.