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Volume 21, Issue 3, Pages 367-377 (September 2004) A Direct Role for NKG2D/MICA Interaction in Villous Atrophy during Celiac Disease  Sophie Hüe, Jean-Jacques Mention, Renato C. Monteiro, ShaoLing Zhang, Christophe Cellier, Jacques Schmitz, Virginie Verkarre, Nassima Fodil, Seiamak Bahram, Nadine Cerf-Bensussan, Sophie Caillat-Zucman  Immunity  Volume 21, Issue 3, Pages 367-377 (September 2004) DOI: 10.1016/j.immuni.2004.06.018

Figure 1 MICA Protein Expression Is Increased in Small Intestinal Mucosa of CD Patients Paraffin-embedded biopsy sections from a non-CD control (A), active CD patient (B), CD patient on a gluten-free diet (C), and patient with RCS (D). Magnification 100×. Immunity 2004 21, 367-377DOI: (10.1016/j.immuni.2004.06.018)

Figure 2 Induction of MIC at the Surface of Gut Epithelial Cells and Expression of NKG2D on Intraepithelial Lymphocytes in Active CD and RCS Flow cytometry analysis of epithelial cells and IELs freshly isolated from duodenal biopsies in control subjects (n = 4), treated CD (n = 5), active CD (n = 4), and RCS (n = 4) patients. The numbers indicate the percentage of gated cells. A representative experiment from each group is shown. Left: ESA-positive epithelial cells stained with PE-labeled anti-MICA SR116 mAb. In the upper right quadrant, intracellular staining of permeabilized cells is shown in the same control subject. Right: CD2+ CD7+ IELs stained with FITC-labeled MICA tetramers. In parallel to IELs, staining of PBMC is shown in a treated patient. Immunity 2004 21, 367-377DOI: (10.1016/j.immuni.2004.06.018)

Figure 3 MICA Expression Is Induced by Gliadin or Gliadin-Derived Peptides in the Small Intestine Mucosa of CD Patients Biopsy specimens from healthy controls (A, C, E, G, and I) and CD patients on a gluten-free diets (B, D, F, H, J, and K–N) were incubated for 12 hr in medium alone (A and B) or in presence of peptic-tryptic digest of gliadin (C and D) or p31-49 gliadin peptide (E and F), p57-89 gliadin peptide (G and H), IL-15 (I and J), p31-49 plus anti-IL-15 neutralizing mAb (K), p57-89 plus anti-IL-15 mAb (L), peptic-tryptic digest of BSA (M), and hSA control peptide (N) prior to staining with SR99 anti-MIC mAb. In (A) and (N), weak nonspecific staining of red blood cells is observed in some lamina propria blood vessels. Magnification 160×. Immunity 2004 21, 367-377DOI: (10.1016/j.immuni.2004.06.018)

Figure 4 NKG2D-Mediated Cytotoxicity of IELs in Active CD and RCS (A–C) Redirected cytotoxicy of FcγR+ P815 cells by IEL cell lines. Results represent triplicate of lysis for each cell line in a given experiment. (A) IEL cell lines from five different active CD patients are tested at a 30:1 effector:target cell ratio, in the presence of anti-CD3 mAb (0.01 ng/ml) and/or anti-NKG2D mAb (1 μg/ml), mAb, or isotype control IgG1. (B) IEL cell line from one representative patient with active CD, tested at different kinetic time points of the cell culture (day 14, 15, 19, and 26), shows transient capacity in the NKG2D-mediated killing of P815 cells. (C) NKG2D-mediated killing of P815 cells by IEL cell lines from four patients with RCS in the presence of control IgG1 or anti-NKG2D mAb (effector:target cell ratio 10:1). Lysis is fully abrogated by the PI3K inhibitor LY 294002 (patient BER, n = 4). (D and E) Direct cytotoxicity of MICA-positive epithelial cells by IEL cell lines. (D) Cytotoxicity of HT29 epithelial cells by IEL cell line from RCS patient BER (10:1 E:T ratio) in presence or absence of isotype control IgG1, anti-MICA, or anti-NKG2D mAb or LY294002. (E) IEL cell lines from five active CD patients show variable direct cytotoxicity of HeLa cells at high effector:cell ratio (100:1). Lysis is only weakly abrogated by anti-NKG2D mAb. Immunity 2004 21, 367-377DOI: (10.1016/j.immuni.2004.06.018)

Figure 5 Soluble MICA in the Serum of CD Patients Does Not Downmodulate NKG2D Levels on IELs (A) Soluble MICA is present in the serum of CD patients. Serum samples from 24 healthy individuals, 22 CD patients on gluten-free diets, and 32 untreated CD patients were investigated by ELISA with recombinant sMICA*008 as a standard. The data shown are means of triplicates. Horizontal lines indicate mean value of respective groups. (B) Failure of soluble recombinant MICA to downmodulate NKG2D on IEL lines from active CD or RCS patients. Two-color staining of IEL cells incubated in the absence or presence of soluble recombinant MICA (10 μg/ml) with or without IL-15 (20 ng/ml). Data are representative of staining on three different IEL lines. Immunity 2004 21, 367-377DOI: (10.1016/j.immuni.2004.06.018)