Thresholds of serum S100A9 concentration able to identify non-responders patients.

<p>Absolute quantification by ELISA of serum S100A9 protein in responders versus non-responders prior to MTX/ETA initiation. The calculated thresholds resulting from ROC analyses are also given with the corresponding sensitivities (Sen) and specificities (Spe). Circles and squares represent individual points for R (n = 12) and NR (n = 10) patients respectively.</p

Baseline expression of S100A8 and S100A9 in PBMCs from RA patients according to the response/non-response status to the ETA/MTX combination in the first population.

<p>Relative quantification by mass spectrometry of S100A9 protein (A) and S100A8 protein (B) at baseline in responders (n = 3) versus non-responders (n = 3) to MTX/etanercept (significant difference is noted by asterisk, p<0.05; Mann-Whitney non-parametric test). Both S100A9 and S100A8 proteins accumulated in R patient (p-value  = 0.0022). The upper and lower bounds of each box indicate the 25^th and 75^th percentile respectively and heavy lines within the box represent the median. Whiskers are drawn to the min and max values.</p

Over-expression of the S100A9 protein in baseline serum from responder patients to the MTX/ETA combination according to 2 different approaches in the second population.

<p>Relative quantification of serum S100A9 protein at baseline in R (n = 12) versus NR (n = 10) by mass spectrometry (A). This result showed an over-expression of S100A9 protein in R patient (p-value  = 0.0022). Serum ELISA absolute quantification of S100A9 (B), S100A8 (C) and calprotectin (D) at baseline in responders versus non-responders. No significant difference in the expression of S100A8 protein and calprotectin was observed between R and NR (p-value  = 0.75 and 0.32, respectively). Only S100A9 showed an over-expression in R patients (p-value  = 0.023).</p

Demographic, clinical and biological data of RA patients at baseline.

<p>Demographic, clinical and biological data of RA patients at baseline.</p

Relationship between baseline levels of several potential markers measured prior to MTX/ETA initiation and the degree of response over the 6 months follow-up in the second population.

<p>Relationship between baseline levels of several potential markers measured prior to MTX/ETA initiation and the degree of response over the 6 months follow-up in the second population.</p

Experimental workflow.

<p>In the first stage, a label free approach was used to determine candidate proteins that are differentially expressed in PBMCs between responder (R) and non-responder (NR) RA patients before initiation of etanercept in the population 1. This approach was used to quantify the relative abundance of S100 proteins. To independently confirm these results, label free approach by mass spectrometry and absolute quantification by ELISA were used to quantify S100 proteins in sera samples from R and NR RA patient before etanercept initiation in the population 2.</p

Additional file 6: of Dysregulation of RasGRP1 in rheumatoid arthritis and modulation of RasGRP3 as a biomarker of TNFα inhibitors

TNF receptor (TNFR)1 and TNFR2 have no effect on RasGRP1 and RasGRP3 gene expression level in T and B cells respectively. Quantitative PCR analysis was performed to measure RasGRP1 and RasGRP3 gene expression levels in T and B cells respectively from three healthy controls. T or B cells were exposed to anti-TNFR1 or anti-TNFR2 neutralizing antibodies without TNFα for 48 hours. The relative expression levels (in arbitrary units (AU)) of RasGRP1 and RasGRP3 were normalized with 18S RNA abundance. Mean ± standard error of the mean were compared using one-way analysis of variance followed by Bonferroni post-hoc test. (TIF 47 kb

Additional file 2: Table S2. of Dysregulation of RasGRP1 in rheumatoid arthritis and modulation of RasGRP3 as a biomarker of TNFα inhibitors

Clinical features of patients and healthy controls. (DOC 40 kb

Additional file 7: of Dysregulation of RasGRP1 in rheumatoid arthritis and modulation of RasGRP3 as a biomarker of TNFα inhibitors

TNFα has no effect on T cell apoptosis. After T cell negative selection, cells were cultured with or without TNFα for 48 and 72 hours. To measure apoptosis, a fluorescein isothiocyanate (FITC) annexin-V Apoptosis Detection Kit (BD Biosciences, USA) was used. After labeling with FITC annexin-V and propidium iodide (PI), cells were analyzed by flow cytometry within 1 hour. a One representative result of three independent experiments is presented. b Histogram represents the mean ± standard error of the mean (SEM) of percentage of annexin-V positive cells in three independent experiments. Mean ± SEM were compared using Student’s t test. (TIF 1318 kb

No difference in serum levels of IL-4 and IL-6 in ENO1 or pEP1 treated mice compared to control mice.

<p>Serum levels of IL-4 (A and C) and IL-6 (B and D) in ENO1 treated mice (A-B) and pEP1 treated mice (C-D), compared to control mice, were measured by Luminex. Bars represent the mean ± SEM. Statistical analysis used are 2way ANOVA (A and B) or Mann-Withney (C and D).</p