Immunohistochemistry of an accessory salivary gland in Sjögren syndrome.

<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

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

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

Surface phenotype of CD4 T cells with respect to NKG2D and CD28.

<p>This, more global figure, analyses the ratio of CD4⁺ T cells harbouring or not CD28 and/or NKG2D, or not. None of the differences is statistically significant.</p

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

<p>The panel represents RNA originating from major human organs. Each panel contains lung RNA as an internal standard. Top panel was hybridized with <i>MICA</i> cDNA. The differential transcript length of <i>MICA</i> and <i>MICB</i> is due to a previously documented larger 3′UT in <i>MICB</i> mRNA <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000518#pone.0000518-Bahram4" target="_blank">[50]</a>. The middle panel depicts the expression of <i>β₂m</i>, indicative of the β₂m bound conventional MHC-I expression. Finally the <i>β-actin</i> transcription acts as a loading control. For more detailed explanation see main text.</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

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

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