elevated level of IL-1β and IL-23 secreted by MoDCs of CAPS patients upon Zymosan stimulation.

<p>Secretion of IL-1β (A), IL-23 (B) and IL-6 (C) by MoDs upon 48 hours of challenge with or without Zymosan in 4 CAPS patients, 10 healthy controls and 4 active SoJIA patients. Bold horizontal lines represent median values. Boxes contain the 50% of values falling between the 25th and 75th percentiles, whiskers lines that extend from the boxes represent the highest and lowest values for each subgroups. Statistical analysis was performed using non-parametric U Mann-Whitney test.</p

Effect of anti-IL-1 treatment on IL-17 and IL-6 serum levels and Th17 frequency.

<p>Variation of IL-17 (panel A) and IL-6 (panel B) serum levels after anti-IL-1 treatment in 8 CAPS patients and 11 SoJIA patients (7 responders and 5 non responders). The decrease frequency of IL-17 producing T cells after anti-IL-1 treatment is reported either as absolute number (Panel C) and percentage of alive lymphocytes (Panel D). Statistical analysis was performed using non-parametric Wilcoxon-Test.</p

Clinical characteristics of CAPS and SoJIA patients at the moment of the study.

<p>Clinical characteristics of CAPS and SoJIA patients at the moment of the study.</p

Analysis of IL-17 and IL-6 serum concentrations and peripheral CD4+ T cell phenotype.

<p>IL-17 (panel A) and IL-6 (panel B) serum levels were measured in 10 active CAPS patients, 20 active SoJIA patients and 20 healthy controls by ELISA. C–D) <i>Ex-vivo</i> analysis of circulating CCR6⁺ (panel B) and CD161⁺ (panel C) memory T cells (CD4⁺CD45RA⁻) in the same three subgroups. Heterogeneity test among groups was evaluated using the non parametric Kruskal-Wallis test (upper right of each graph). Post-hoc analysis with non-parametric U Mann-Whitney test revealed the difference among the three subgroups.</p

CAPS patients show an higher absolute number of IL-17 producing cells after <i>in vitro</i> expansion.

<p>A) Dot plot electronically gated on alive CD4⁺ T cells of one representative CINCA patient, one systemic onset juvenile idiopathic arthritis (SoJIA) patient and one healthy control B–C) Absolute number of IL-17 (panel B) and IFN-γ (panel C) producing cells in 7 NLPR3-mutated CAPS and 12 SoJIA active patients and in 9 age-matched healthy controls. Heterogeneity test among groups was evaluated using the non parametric Kruskal-Wallis test (upper right of each graph). Post-hoc analysis with non-parametric U Mann-Whitney test revealed the difference among the three subgroups.</p

Expression of CXCR3 and interferon (IFN)-γ by (SF) CCR7and CCR7memory CD4cells from synovial fluid

<p><b>Copyright information:</b></p><p>Taken from "Phenotypic and functional characterisation of CCR7and CCR7CD4memory T cells homing to the joints in juvenile idiopathic arthritis"</p><p>Arthritis Research & Therapy 2005;7(2):R256-R267.</p><p>Published online 12 Jan 2005</p><p>PMCID:PMC1065323.</p><p>Copyright © 2005 Gattorno et al.; licensee BioMed Central Ltd.</p> IFN-γ expression was investigated by three-colour staining of freshly isolated SF CD45ROcells with CD4–fluorescein isothiocyanate (FITC), anti-CCR7–phycoerythrin (PE) and anti-CCR5–CyChrome monoclonal antibodies (mAbs) or CD4–TC (where TC stands for Tri-color), anti-CCR7–PE and anti-IFN-γ mAbs, respectively; CXCR3 expression was investigated by triple staining with CD4–TC, anti-CCR7–PE and anti-CXCR3–FITC, as described in the Methods section. Subsequently, cytofluorimetric analysis was performed by gating on the CD4CCR7and CD4CCR7lymphocyte subsets. Data are expressed as percentages of positive cells or/and mean fluorescence intensity. Expression of IFN-γ (a) and CXCR3 (b) by SF CCR7and CCR7memory CD4cells from 10 patients with juvenile idiopathic arthritis (JIA). Boxes contain values falling between the 25th and 75th centiles; whiskers show lines that extend from the boxes represent the highest and lowest values for each subgroup. Differences between paired SF mononuclear cells were evaluated by the Wilcoxon rank test. Dot plots show the cytofluorimetric analysis IFN-γ (c) and CXCR3 (d) expression by the gated CD4CCR7(gate 1) and CD4CCR7(gate 2) cell populations in three representative patients with JIA

Additional file 1: Table S1. of Patient’s experiences with the care for juvenile idiopathic arthritis across Europe

Number of questionnaires per country and division into regions. Table S2. SHARE PARENT SURVEY: Patient Personal Information and Diagnosis. (DOCX 24 kb

Expression of LAIR-1 and NKp44 on peripheral tissue pDCs.

<p>pDCs were identified in different tissues as BDCA-2⁺ ILT3⁺ positive cells (A). LAIR-1 and NKp44 expression was evaluated in tonsils (B), lymph nodes (C and E) and in different malignant tumors (D and E). pDCs activation in lymph nodes and in NSCLC was assessed as CD83 expression (E and F). The horizontal bars within dot-plots of panels C, D, F represent negative controls with isotype-matched irrelevant mAbs. Data shown are representative of at least three independent experiments analyzing pDCs from different patient tissues.</p

LAIR-1 triggering on pDCs induces inhibition of IFNα production.

<p>LAIR-1 cross-linking inhibits IFNα production by pDCs activated with either CpG (A) or DNA immunocomplexes (B). pDCs were cultured for 18 h in the presence of IL-3, incubated with the indicated mAbs or isotype-matched mAbs (Ctrl mAb) and then cultured in F(ab)2 goat anti-mouse IgG antibody-coated plates in the presence of CpG or DNA immunocomplexes. IFNα production was assessed by ELISA in culture supernatants. Graphs represent mean values +/− SEM of three independent experiments. ** p<0.01; * p<0.05.</p

Expression of LAIR-1 and NKp44 on peripheral blood pDCs from healthy donors.

<p>Expression of LAIR-1 was assessed on pDCs from healthy donors PBMCs (A) and on purified pDCs (B) following culture in the presence of IL-3 (20 ng/ml), CpG (5 µg/ml) or IFNα (1000 U/ml) for 48 hours. (B): Dotted lines depict unstimulated cells; bold lines depict cells cultured as indicated. Values represent expression Geo-MFI. All experiments related to the analysis of LAIR-1 expression on pDCs cultured with the indicated stimuli for 24 and 48 hours are summarized in (C). Values depict Geo-MFI of positive cells/Geo-MFI of Ig isotype-matched control stained cells. (D): Analysis of LAIR-1, NKp44 and DAP12 mRNA expression on human pDCs. Polyclonal NK cell lines, pDCs and pDCs cultured 48 h with IL-3 were analyzed by RT-PCR for LAIR-1, NKp44 and DAP12 expression (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0015080#s4" target="_blank">Materials and Methods</a>).</p