Table 1.
Main demographic features of MS patients and healthy control groups.
Fig 1.
GM-CSF mRNA and GM-CSF-producing cells in PBMCs from relapsing and stable MS patients and from healthy controls (HC).
Quantitative-PCR to measure GM-CSF mRNA expression (A) ex vivo (Relapsing MS: n = 30, Stable MS: n = 16, HC: n = 18) and (B) after 4h of stimulation by PMA/ionomycin (Relapsing MS: n = 14, Stable MS: n = 10, HC: n = 12). (C) PBMCs were stained for GM-CSF, CD3 and CD4 after 4h stimulation by PMA/ionomycin in the presence of a protein transport inhibitor and analysed by flow cytometry (Relapsing MS: n = 15, Stable MS: n = 12, HC: n = 15). Scatter dot plot illustrates the percentage of GM-CSF+CD4+ cells. The horizontal lines of scatter plots represent the median value in all subgroups.
Table 2.
GM-CSF expression analysis in PBMCs of MS patients and HC.
Fig 2.
IL-22 mRNA and IL-22-producing cells in PBMCs from relapsing and stable MS patients and from healthy controls (HC).
Quantitative-PCR to measure IL-22 mRNA expression (A) ex vivo (Relapsing MS: n = 56, Stable MS: n = 16, HC: n = 37) and (B) after 4h of stimulation by PMA/ionomycin (Relapsing MS: n = 14, Stable MS: n = 10, HC: n = 12). (C) PBMCs were stained for IL-22, CD3 and CD4 after 4h stimulation by PMA/ionomycin in the presence of a protein transport inhibitor and analysed by flow cytometry (Relapsing MS: n = 15, Stable MS: n = 12, HC: n = 14). Scatter dot plot shows the percentage of IL-22+CD4+ T cells. The horizontal lines of scatter plots represent the median value in all subgroups. * and *** indicate respectively p-values ≤ 0.05 and ≤ 0.001.
Table 3.
IL-22 expression analysis in PBMCs of MS patients and HC.
Fig 3.
FACS analysis of GM-CSF, IL-22 and CD39 coexpression with IL-17 in CD4+ T cells.
PBMCs were fixed and stained after 4h stimulation by PMA/ionomycin in the presence of a protein transport inhibitor. CD4+ cells were gated among the CD3+ population. Representative dot plots of (A) GM-CSF, (B) IL-22 and (C) CD39 costaining with IL-17. Numbers in each quadrant represent the average percentage of positive CD4+ T cells in relapsing MS patients.
Table 4.
Characterisation of GM-CSF, IL-22 and CD39 expression by Th17 lymphocytes during MS relapses.
Fig 4.
GM-CSF and IL-22 before and after intravenous methylprednisolone (ivMP) treatment in paired samples of relapsing MS patients.
Quantitative-PCR to measure GM-CSF and IL-22 mRNA expression ex vivo (A, n = 13 and D, n = 17) and (B, n = 8 and E, n = 8) after 4h stimulation by PMA/ionomycin. Results are expressed relative to the mean of the relapsing patients set at 1. PBMCs were stained for GM-CSF (C, n = 8) or IL-22 (F, n = 15), CD3 and CD4 after 4h stimulation by PMA/ionomycin in the presence of a protein transport inhibitor and analysed by flow cytometry. The horizontal lines of scatter plots represent the median value in all subgroups. (G) PBMCs of HC (n = 5) were treated with methylprednisolone (MP) for 24h and stimulated 2h with PHA. Error bars represent the standard error of the mean. * and *** indicate respectively p-values ≤ 0.05 and ≤ 0.001.