Expression analysis of <i>MICA</i> and <i>MICB</i> by real-time quantitative PCR in HeLa cells.

<p><i>MICA</i> (A) and <i>MICB</i> (B) expression profile as analyzed by RT-qPCR. ▪ Samples 15 minutes after heat shock, ♦ Samples 60 minutes after heat shock, ▴ untreated controls.</p

Characteristics of patients and healthy controls.

*<p>DAS (Disease Activity Score) 28 for RA, SLEDAI (Systemic Lupus Erythematosus Disease Activity Index) for SLE and mean AVS (AVS: Analogic Visual Scale) of subjective sicca syndrome, asthenia and pain for SS.</p>**<p>Abbreviations: NSAID: Non-Steroidal Anti-Inflammatory Drugs; SMARD: Symptom Modifying Anti-Rheumatic Drugs; DCART: Disease Controlling Anti-Rheumatic Therapy; ANA: antinuclear antibody (indirect immunofluorescence on Hep2 cells), ENA: extractable nuclear antigens; dsDNA: double-stranded DNA, RF: Rheumatoid Factor, CCP: cyclic citrullinated peptide.</p

CD4⁺NKG2D⁺ T cells autoimmune diseases.

<p>This figure depicts the ratio of CD4⁺NKG2D⁺ T cells within the total peripheral CD4 pool in control, RA, SLE and SS individuals. The variability in SS patients is due to 2 individuals whom represent an unusually high ratio (see main text for explanation). None of the differences is significant as assessed by the Wilcoxon text: p = 0.1658 for controls vs. RA; p = 0.9547 for controls vs. SLE and p = 0.5012 for controls vs. SS.</p

Expression analysis of <i>MICA</i> and <i>MICB</i> by real-time quantitative PCR in Sjögren's syndrome accessory salivary glands.

<p>Relative expression level of <i>MICA</i> (A) and <i>MICB</i> (B) in Sjögren's syndrome patients as compared to healthy controls. The differences between patients and controls are not significant as assessed by Mann-Whitney <i>U</i> test (α = 0.27 for <i>MICA</i> and α = 0.53 for <i>MICB</i>).</p

Immunohistochemistry of RA joint infiltrate.

<p>(A) Color stain by Hematoxylin-eosin staining. Immunohistochemical analysis using (B) CD4 (C) CD8 and (D) NKG2D antibodies. The infiltrate is clearly of lymphocytic origin with a majority of CD4 T cells.</p

MICA expression in Rheumatoid Arthritis synovitis and Sjögren syndrome accessory salivary glands.

<p>Immunohistochemical analysis of (A) (B) RA synovitis and an (C) (D) SS accessory salivary gland stained with an anti-MICA monoclonal antibody. MIC expression is clearly of epithelial/fibroblastic nature in both tissues/organs.</p

Flow cytometric analysis of the CD4⁺NKG2D⁺ population.

<p>This example, taken from one of the two SS patients (Mrs XET) harbouring a particularly high ratio of CD4⁺NKG2D⁺ T cells (36.2%) illustrates the methodology used in order to enumerate these cells in all individuals (controls and patients) included in this work during a routine 50 000 events analysis.</p

Northern blot analysis of <i>MICA</i> and <i>MICB</i> expression in various tumors.

<p>The same disposition as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000518#pone-0000518-g001" target="_blank">Figure 1</a> applies, but this time the blots contain tumor and control (adjacent disease-free tissue) lanes.</p

Deregulation of Type I IFN-Dependent Genes Correlates with Increased Susceptibility to Cytomegalovirus Acute Infection of Dicer Mutant Mice

<div><p>Regulation of gene expression by microRNAs (miRNAs) is now considered as an essential mechanism for cell development and homeostasis. Indeed, numerous studies have reported that modulating their expression, maturation, or activity can affect cell survival, identity or activation. In particular, miRNAs are key players in the tight regulation of signaling cascades, and as such, they appear as perfectly suited immunomodulators. Several immune-related processes, including inflammation, have recently been demonstrated to require specific miRNAs. In addition, the discovery of herpesvirus-encoded miRNAs has reinforced this assumption. To decipher the potential roles of miRNAs in innate antiviral immune response, we developed an <em>in vivo</em> model based on the inoculation of mouse cytomegalovirus (MCMV) in mice. Furthermore, we exploited a mouse line carrying a hypomorphic mutation in the <em>Dicer</em> gene to visualize the impact of impaired miRNA biogenesis upon the anti-MCMV response. Our data indicate that miRNAs are important actors in mounting an efficient response against herpesviruses. We suggest that a rapid and transient interferon response following viral infection requires miRNA-dependent repressor release. In addition, our <em>in vivo</em> efforts identified several miRNA targets, thus providing a conceptual framework for future analyzes on the regulation of specific actors involved in the Type I interferon pathway.</p> </div

IFN-stimulated genes induction is impaired in MCMV-infected <i>Dicer^d/d</i> mice.

<p>A. Scatterplot shows multiple IFN-dependent genes up-regulation (red dots) in MCMV-infected control mice (+/+). B. Scatterplot analysis comparing MCMV-infected controls (+/+) and <i>Dicer^d/d</i> mice. C–E. qRT-PCR data on selected MCMV-inducible genes (<i>Irf7, Ifit1</i> and <i>Oas1a</i>). The graphs show relative gene expression in controls (black spots; +/+) and <i>Dicer^d/d</i> (white spots) spleen extracts. F. CXCL10 was measured in the serum of controls (white dots, +/+) and <i>Dicer^d/d</i> mice (black dots) 36 hours p.i. G. IRF7 expression was analyzed by Western blot performed on protein extracts from MCMV-infected wild-type (+/+) and mutant mice. GAPDH expression serves a loading control.</p