1. Volume 136, Issue 7, Pages 2270-2279 (June 2009)
Interleukin-25 Inhibits Interleukin-12 Production and Th1 Cell-Driven Inflammation in the Gut 
Roberta Caruso, Massimiliano Sarra, Carmine Stolfi, Angelamaria Rizzo, Daniele Fina, Massimo Claudio Fantini, Francesco Pallone, Thomas T. MacDonald, Giovanni Monteleone 
Gastroenterology 
Volume 136, Issue 7, Pages (June 2009)
DOI: /j.gastro
Copyright © 2009 AGA Institute Terms and Conditions

2. Figure 1
IL-25R is expressed by mucosal CD14+ cells in Crohn's disease. (A and B) Percentages of CD14+ (A) and CD3+ (B) cells positive for IL-25R in LPMC isolated from patients with Crohn's disease (CD), ulcerative colitis (UC), and normal controls (CTR). Data indicate mean ± SD of 5 separate experiments in which LPMC isolated from 5 CD patients, 5 UC patients, and 5 CTR were analyzed. Right insets in panel A show representative dot plots of IL-25R and CD14 in LPMC isolated from 1 patient with CD. Numbers in quadrants indicate the percentages of CD14+ cells positive for IL-25R. Staining with a nonrelevant isotype IgG is also shown. (C–E) CD14+ cells were isolated from the inflamed gut of 5 patients with CD and preincubated with medium or IL-25 for 30 minutes then either left unstimulated (M, medium) or stimulated with LPS or PGN for 48 hours. Cell culture supernatants were then collected and analyzed for IL-12p70 (C), IL-23 (D), and IL-10 (E) by ELISA. Data indicate mean ± SD of all experiments. *P < .01, **P = .03.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

3. Figure 2
(A) Percentages of CD11b+, CD11c+, and F4/80+ cells positive for IL-25R in LPMC isolated from the colons of mice treated with ethanol (ETOH) or PGN. Five mice were treated with ETOH, and 5 were treated with PGN. At day 4, mice were killed, LPMC isolated and pooled, and then analyzed by flow-cytometry. (B) IL-25 largely prevents PGN-mediated colitis. IL-25 (10 μg/mouse) was injected intraperitoneally 1 hour before the administration of PGN, and body weight was recorded daily. Data indicate mean ± SEM of separate experiments. In each experiment, each group consisted of at least 5 mice. PGN vs PGN+IL-25, *P = .02. (C) Histologic score of the colon sections taken from control (ETOH) and colitic mice either treated with or without IL-25. Data indicate mean ± SD of 3 separate experiments. For each experiment, at least 5 mice per group were considered. *P = .01. (D–F) Administration of IL-25 to mice with PGN-colitis results in decreased tissue expression of IL-12p70 (D), IL-12p40 (E), and IFN-γ (F). Data indicate mean ± SD of all experiments (*P = .03, **P = .01).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

4. Figure 3
The IL-25-mediated prevention of PGN-colitis is not mediated by IL-13. (A) Administration of IL-25 to mice with PGN-colitis results in enhanced tissue expression of IL-13. One hour before PGN-colitis induction, mice were either left untreated or treated with IL-25 (10 μg/mouse). Mice were killed at day 5, and colonic extracts were analyzed by ELISA. Data indicate mean ± SD of all experiments (*P = .02). (B) Balb/c mice were injected intraperitoneally with a neutralizing IL-13 or control IgG and 1 hour later with murine IL-25 followed by PGN as indicated in Materials and Methods section. Upon autopsy on day 5, the colons were removed, and the length was measured; representative colons are shown. (C) Histologic score of the colon sections taken from colitic mice treated as indicated in A. Data indicate the mean ± SD of all experiments in which at least 3 mice per group were considered.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

5. Figure 4
IL-25 largely reverses PGN-colitis. (A) IL-25 was injected intraperitoneally the day after the induction of PGN-colitis. Body weight changes of control and colitic mice either untreated or treated with IL-25 were daily recorded. Data indicate mean ± SEM of 2 separate experiments. In each experiment, each group consisted of at least 6 mice. PGN-treated vs PGN+IL-25-treated mice, *P = (B) Representative H&E-stained sections of colon from mice with PGN-colitis given PBS or treated with IL-25. Right inset shows the histologic score of the colon sections taken from colitic mice either left untreated or treated with IL-25. Data indicate the mean ± SD of all experiments in which at least 6 mice per group were considered. Giving IL-25 to mice with PGN-colitis results in decreased tissue expression of IL-12p70 (C), IL-12p40 (D), and IFN-γ (E). Data indicate mean ± SD of all experiments (*P < .02).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

6. Figure 5
IL-25 ameliorates TNBS-mediated colitis. IL-25 was injected intraperitoneal the day after the induction of TNBS-colitis. (A) Photomicrograph (40×) of an H&E-stained paraffin section of representative colonic sections from mice belonging to each group. Severe mucosal mononuclear cell infiltrate and disruption of the normal crypt architecture with epithelial ulceration and loss of goblet cells are evident in the colon of mice with TNBS colitis. In contrast, only a minimal cellular infiltration of the mucosa is seen in the colon of mice treated with IL-25. The photomicrographs are representative of 3 separate experiments in which at least 5 mice per group were studied. Right inset: Histologic score of the colon sections taken from control (ETOH) and colitic mice either treated with or without IL-25. Data indicate mean ± SD of all experiments. *P = .01. (B–D). Giving IL-25 to mice with TNBS-colitis results in decreased tissue expression of IL-12p70 (B), IL-12p40 (C), and IFN-γ (D). Data indicate mean ± SD of all experiments. *P < .03.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

7. Figure 6
IL-25 ameliorates oxazolone-induced colitis. IL-25 was injected intraperitoneally the day after the induction of oxazolone-colitis. (A) Body weight changes of control (ETOH) and colitic mice either untreated or treated with IL-25 were daily recorded. Data indicate mean ± SEM of 2 separate experiments. In each experiment, each group consisted of at least 4 mice. Oxazolone-treated vs oxazolone+IL-25-treated mice, *P = .03. (B) Upon autopsy on day 5, the colons were removed, and the length was measured; representative colons are shown. (C) Photomicrograph (40×) of an H&E-stained paraffin section of representative colonic sections from mice belonging to each group. Severe mononuclear cell infiltrate involving the whole colonic wall, with disruption of the normal crypt architecture and loss of goblet cells is evident in the colon of mice with oxazolone colitis. In contrast, only a minimal cellular infiltration of the mucosa is seen in the colon of mice treated with IL-25. The photomicrographs are representative of 2 separate experiments in which at least 4 mice per group were studied.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions

8. Figure 7
IL-25 expression is down-regulated in the inflamed colon of patients with IBD. (A) Real-time PCR data for IL-25 RNA transcripts in colonic biopsy specimens of 10 normal controls (CTR), 10 patients with active UC, and 10 patients with active CD. Levels are normalized to β-actin and indicate the mean ± SD of all experiments. *P = .01. Right inset shows real-time PCR data for IL-25 RNA transcripts in colonic biopsy specimens of 5 patients with IBD before (B) and after (A) the medically induced remission. *P = .03. (B) Representative Western blots showing IL-25 and β-actin protein in mucosal samples taken from 2 CTR, 2 patients with UC, and 2 patients with CD. One of 4 separate experiments analyzing in total samples of 9 CTR, 10 UC patients, and 9 CD patients is shown. (C) Quantitative analysis of IL-25/β-actin protein ratio in mucosal samples taken from 9 CTR, 10 UC patients, and 9 CD patients as measured by densitometry scanning of Western blots. Values are expressed in arbitrary units (a. u.). Each point represents the value of IL-25/β-actin protein ratio in mucosal samples taken from a single subject. Horizontal bars indicate the median. *P = (D) IL-25 immunostaining in intestinal specimens of 1 patient with CD, 1 patient with UC, and 1 CTR. The Figure is representative of 5 separate experiments in which sections of 5 patients with CD, 5 patients with UC, and 5 normal controls were analyzed. Original magnification, 40× (100× in the right lower inset of CTR picture).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2009 AGA Institute Terms and Conditions