1. Volume 136, Issue 2, Pages 530-541 (February 2009)
Glucocorticoid-Induced Leucine Zipper Is Protective in Th1-Mediated Models of Colitis 
Lorenza Cannarile, Salvatore Cuzzocrea, Luca Santucci, Massimiliano Agostini, Emanuela Mazzon, Emanuela Esposito, Carmelo Muià, Maddalena Coppo, Rosanna Di Paola, Carlo Riccardi 
Gastroenterology 
Volume 136, Issue 2, Pages (February 2009)
DOI: /j.gastro
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2. Figure 1
GILZ-TG mice are less susceptible than WT mice to DNBS-induced colitis. Four and 7 days after DNBS administration, mice were killed and body weight (A) and macroscopic damage scores (B) evaluated. (Data are the means ± SE of 10 mice for each group). Representative photographs of colon from sham WT mice (C), colon from DNBS-treated WT mice (D), and colon from DNBS-treated GILZ-TG mice (E). Bars indicate means ± SE. *P < .01, DNBS-treated vs vehicle; °P < .01, DNBS-treated GILZ-TG vs DNBS-treated WT.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

3. Figure 2
Reduced colon injury (A) and MPO activity (C) in DNBS-treated GILZ-TG as compared with DNBS-treated WT mice. (A) H&E staining of colons from WT and GILZ-TG mice, killed 4 days after intrarectal administration of DNBS. Colon from WT mice shows mononuclear cell infiltration with loss of goblet cells. Less cell infiltration and histologic alteration were observed in GILZ-TG mice. (B and C) Histologic damage score and colonic MPO activity were evaluated. Bars indicate means ± SE. *P < .01, DNBS-treated vs vehicle; °P < .01, DNBS-treated GILZ-TG vs DNBS-treated WT.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

4. Figure 3
P-selectin, ICAM-1, FasL expression, and TUNEL assay in the colon of DNBS-treated WT and GILZ-TG mice. Positive staining for P-selectin (A), ICAM-1 (B), TUNEL (C), and FasL (D) in colons from DNBS-treated WT mice compared with DNBS-treated GILZ-TG and sham controls. No positive staining for P-selectin, ICAM-1, and FasL in colons from DNBS-treated GILZ-TG mice. More apoptotic cells, evaluated by TUNEL assay, were detected in DNBS-treated WT.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

5. Figure 4
Colonic infiltration of CD4+ cells and cytokine production. (A and B) Infiltration of CD4+ T cells and IL-2 production in colons isolated from WT and GILZ-TG mice 4 days after administration of DNBS (arrows indicate positive cells). Markedly less CD4+ T cells and IL-2 were detected in the colon of DNBS-treated GILZ-TG mice in comparison with DNBS-treated WT mice. (C and D) Colonic concentrations of TNF-α and IL-1β as determined by enzyme-linked immunosorbent assay (day 4 after DNBS administration). Data are the means ± SE of 10 mice for each group. *P < .01 vs vehicle; °P < .01 vs DNBS-treated WT mice.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

6. Figure 5
NF-κB activation is lower in colon of DNBS-treated GILZ-TG than in WT mice. At day 4 after DNBS treatment, colons were removed and processed for protein extraction. Western blot analysis of p65NF-κB (A) and phosphorylated p65 on serine 536 (Ser 536, B) shows a significant reduction in NF-κB nuclear levels and phosphorylation of p65 on Ser536 in GILZ-TG as compared with WT mice. Data were quantified and expressed as arbitrary densitometric units. Bars indicate means ± SE. *P < .01, comparing column 3 with column 4.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

7. Figure 6
NF-κB nuclear translocation and cytokine production in LP CD4+ T cells from WT and GILZ-TG DNBS-treated mice. (A) LP CD4+ T cells were purified, treated for 2 hours as indicated, and then processed for immunofluorescence assay using anti-p65 monoclonal antibodies. 4′,6-Diamidino-2-phenylindole (DAPI) staining was used to identify nuclei; p65NF-κB (red) and DAPI (blue) images have been merged. Figure shows a representative experiment of 3 independent experiments with similar results. (B) LPMC (top) and purified CD4+ T cells (bottom) from LP were isolated 4 days after intrarectal administration of DNBS or ethanol alone, were cultured for 48 hour in wells containing immobilized anti-CD3 and soluble anti-CD28 monoclonal antibodies, and cytokine concentrations in the supernatants were determined by enzyme-linked immunosorbent assay (data are the means ± SE of 10 mice for each group). *P < .01 vs WT mice.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

8. Figure 7
TAT-GILZ fusion protein and DEX treatment reduce the clinical, histologic, and immunologic signs of DNBS-induced colitis. Mice were killed 4 days after DNBS administration. Vehicle, TAT, or TAT-GILZ was injected starting 1 day before DNBS administration. (A) Macroscopic observation of the colon tissues from sham-treated mice (1), DNBS-treated mice (2), DNBS-treated mice that have received TAT administration (3), DNBS-treated mice that have received TAT-GILZ treatment (4), and DNBS-treated mice receiving DEX treatment (10 mg/kg intraperitoneally) (5). (B) Effect of TAT-GILZ on body weight change, macroscopic and histologic damage score, and colon MPO activity. *P < .05 vs control; °P < .05 vs colitic mice treated with vehicle or TAT alone. (C) Colon injury was assessed by H&E and trichrome coloration. No histologic alterations were observed in the colon section from sham-treated mice. Mucosal injury characterized by absence of epithelium and a massive mucosal and submucosal infiltration of inflammatory cells was observed in the colon section from DNBS-treated mice that have received TAT administration. Treatment with TAT-GILZ and DEX corrected the morphologic disturbances associated with DNBS administration. (D) Immunocytochemistry analysis of GILZ levels in mucosal T lymphocytes. No positive staining for GILZ was observed in colon from sham-treated mice as well as in the tissue sections from DNBS-treated WT mice. On the contrary, colon sections from DNBS-treated WT mice that have been treated with DEX revealed a positive staining for GILZ, mainly localized in T lymphocytes (see particle).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

9. Figure 8
Administration of TAT-GILZ fusion protein exerts therapeutic activity in fully established colitis in IL-10KO mice. TAT-GILZ (0.2 mg/kg, 3 times per week) or TAT (0.1 mg/kg, 3 times per week) was administered subcutaneously for 7 consecutive weeks, starting at 9 weeks of age. (A) Effect of TAT-GILZ administration on histologic score of colitis, colonic MPO activity, and colonic MIP-1α and TNFα levels. *P < .01 vs 16-week-old WT mice; *°P < .01 vs 9-week-old IL-10KO mice; *°°P < .05 vs 16-week-old IL-10KO mice. (B) H&E staining of colon from 9-week-old IL-10KO mouse (note the presence of a fully established colitis), 16-week-old IL-10WT mouse (no colitis), and 16-week-old IL-10KO mice treated with TAT (thickening of the colon wall and massive inflammatory infiltrate in the LP) or TAT-GILZ (subepithelial edema with no inflammatory infiltrate).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions