Table S2. Comparison of fold change of protein phosphorylation between MS patients and controls after in vitro stimulation. Fold change in the levels phosphorylated proteins induced by in vitro stimulation in each cell type in healthy controls and RRMS patients. Values represent the mean fold change of phosphorylation levels and standard deviation for each group. (DOCX 15Â kb

Alessandro Didonna

Ester Canto

Jorge R. Oksenberg

MS-EPIC Study Group

Noriko Isobe

Stephen L. Hauser

Journal of Neuroinflammation

Table S4. Comparison of levels of p38MAPK, Erk1/2, STAT1, and STAT6 between MS patients and controls. Levels of selected proteins in each cell type in healthy controls and RRMS patients. Values represent the mean fluorescence intensity and standard deviation for each group. (DOCX 13Â kb

Ester Canto (4920835)

Noriko Isobe (119848)

Alessandro Didonna (4920832)

Stephen Hauser (591884)

Jorge Oksenberg (488201)

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Additional file 5: of Aberrant STAT phosphorylation signaling in peripheral blood mononuclear cells from multiple sclerosis patients

Table S5. Comparison of levels of phosphorylated proteins between RRMS and SPMS patients in baseline conditions. Levels of phosphorylated proteins in each cell type in RRMS and SPMS patients. Values represent the mean fluorescence intensity and standard deviation for each group. (DOCX 14Â kb

Additional file 6: of Aberrant STAT phosphorylation signaling in peripheral blood mononuclear cells from multiple sclerosis patients

Additional file 3: of Aberrant STAT phosphorylation signaling in peripheral blood mononuclear cells from multiple sclerosis patients

Table S6. Comparison of levels of phosphorylated proteins between RRMS and SPMS patients after in vitro stimulation. Levels of phosphorylated proteins in each cell type in RRMS and SPMS patients. Values represent the mean fluorescence intensity and standard deviation for each group. (DOCX 15Â kb

Additional file 7: of Aberrant STAT phosphorylation signaling in peripheral blood mononuclear cells from multiple sclerosis patients

Abstract Background Multiple sclerosis (MS) is characterized by increased activation of peripheral blood mononuclear cells (PBMCs), linked to perturbations in the phosphorylation of signaling proteins. Methods We developed a phosphoflow cytometry protocol to assess the levels of 11 phosphorylated nuclear proteins at baseline conditions and after cell activation in distinct PBMC populations from 41 treatment-naïve relapsing-remitting (RR) MS subjects and 37 healthy controls, and in a second cohort of 9 untreated RRMS patients and 10 secondary progressive (SP) MS patients. Levels of HLA-ABC, HLA-E, and HLA-DR were also assessed. Phosphorylation levels of selected proteins were also assessed in mouse splenocytes isolated from myelin oligodendrocyte glycoprotein (MOG)35–55-induced experimental autoimmune encephalomyelitis (EAE). Results Modest differences were observed at baseline between patients and controls, with general lower phosphorylation levels in cells from affected individuals. Conversely, a dramatic increase in phosphorylated p38MAPK and STAT proteins was observed across all cell types in MS patients compared to controls after in vitro activation. A similar phosphorylation profile was observed in mouse lymphocytes primed in vivo with MOG. Furthermore, levels of all p-STAT proteins were found directly correlated with HLA expression in monocytes. Levels of phosphorylated proteins did not differ between relapsing-remitting and secondary progressive MS patients either in baseline conditions or after stimulation. Lastly, phosphorylation levels appear to be independent of the genotype. Conclusion The response to IFN-α through STAT proteins signaling is strongly dysregulated in MS patients irrespective of disease stage. These findings suggest that the aberrant activation of this pathway could lead to changes in the expression of HLA molecules in antigen presenting cells, which are known to play important roles in the regulation of the immune response in health and disease

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Aberrant STAT phosphorylation signaling in peripheral blood mononuclear cells from multiple sclerosis patients

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Additional file 3: of Aberrant STAT phosphorylation signaling in peripheral blood mononuclear cells from multiple sclerosis patients

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