1. Volume 132, Issue 7, Pages 2359-2370 (June 2007)
Monoclonal Anti–Interleukin 23 Reverses Active Colitis in a T Cell–Mediated Model in Mice 
Charles O. Elson, Yingzi Cong, Casey T. Weaver, Trenton R. Schoeb, Terrill K. McClanahan, Robert B. Fick, Robert A. Kastelein 
Gastroenterology 
Volume 132, Issue 7, Pages (June 2007)
DOI: /j.gastro
Copyright © 2007 AGA Institute Terms and Conditions

2. Figure 1
Colonic tissue cytokine expression. Pathogenic CBA-specific Bir14 T cells or control anti-CD3 activated CD4+ T cells were transferred into groups of 5 C3H.SCID mice. Eight weeks later, colonic tissue RNA was prepared and the expression of multiple genes was analyzed by real-time PCR. (A and B) Expression of cytokine mRNA compared with ubiquitin expression. Each bar represents the mean ± SD of a pool of 5 mice. (C) Cytokines present in supernatants of colon organ cultures at 8 weeks, as measured by enzyme-linked immunosorbent assay.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

3. Figure 2
Th17 cell detection during colitis. (A) Ten days after restimulation with CBA-BMDC, colitogenic Bir14 T cells were stimulated with PMA/ionomycin for 5 hours. Golgi-stop was added in the last 3 hours of culture. Cytokine production was analyzed by intracellular staining. (B) MLN cells from colitic (Bir14 T cell) or control (anti-CD3 T cells) mice were stimulated with PMA/ionomycin for 5 hours, and IL-17 versus IFN-γ production was analyzed by intracellular staining. (C) Lamina propria lymphocytes (LPL) were isolated from control or colitic mice and then stimulated and analyzed as in B. The flow cytometer was gated such that only CD4+ T cells were detected in B and C.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

4. Figure 3
IL-23 promoted but IL-12 inhibited Th17 expansion. MLN CD4+ T cells from colitic C3H IL-10 knockout mice were stimulated with CBA-pulsed APC in the presence of IL-23 (10 ng/mL) or IL-12 (10 ng/mL) for 1 cycle (7 days) and 2 cycles (14 days), and then these T cells were restimulated with PMA/ionomycin for 5 hours and IL-17 and IFN-γ production was analyzed by intracellular staining, gating on total T cells.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

5. Figure 4
Dendritic cell production of IL-12 and IL-23 is antigen and T-cell dependent. Bir14 T cells were cultured with CBA-pulsed BMDCs for 24 hours, or each was cultured alone. IL-12p70 and IL-23 production in the supernatant was measured by enzyme-linked immunosorbent assay. Bars represent mean ± SD of triplicate cultures. CBA-BMDC, BMDCs pulsed overnight with CBA 200 μg protein/mL; OVA-BMDC, BMDCs pulsed overnight with ovalbumin 100 μg protein/mL.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

6. Figure 5
CBA-reactive Th17 cells are potent mediators of colitis. An early passage aliquot of the Bir14 cell line was stimulated with CBA-pulsed APCs in the presence of IL-23 (10 ng/mL), anti–IL-12 (10 μg/mL), and anti–IFN-γ (10 μg/mL) mAb or in the presence of IL-12 (10 ng/mL), IFN-γ (10 ng/mL), and anti–IL-23 (10 μg/mL). After 4 cycles of stimulation, these T cells were restimulated with PMA/ionomycin for 5 hours and IL-17 and IFN-γ production was analyzed by intracellular staining (A). Various numbers of IL-17–producing T cells or IFN-γ–producing T cells were then transferred into groups of 5 C3H.SCID mice. All recipient mice were killed at 8 weeks after cell transfer and histopathology was scored (B). Mean score ± SD at each cell dose is shown. (C) Cytokines detected by enzyme-linked immunosorbent assay in supernatants of colon organ cultures obtained at the time mice were killed from the groups receiving 106 cells (mean ± SD of 5 samples per mouse and 5 mice per group). (D) Representative histopathology at 8 weeks of mice receiving 106 cells, along with a section of normal colon for comparison.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

7. Figure 6
Monoclonal anti–IL-23p19 prevented development of colitis. Bir14 CD4+ T cells were transferred into C3H.SCID mice intravenously, and anti–IL-23p19 mAb or control mAb was administered intraperitoneally (100 μg/mouse) on the same day of cell transfer and weekly thereafter. All recipient mice were killed at 8 weeks after cell transfer, and histopathology was scored. (A) Mean ± SD of quantitative histopathology scores. (B) Representative histopathology of mice receiving control antibody vs anti–IL-23p19 mAb. (C) Colonic tissue RNA was prepared, and the expression of multiple genes was analyzed by real-time PCR. The experiment was performed twice with groups of 4–5 mice each time. *P < .001 vs control.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

8. Figure 7
Anti–IL-23p19 mAb treated active colitis. Bir14 CD4+ T cells were transferred into C3H/HeJ SCID mice intravenously. Four weeks after cell transfer, group 1 was killed and colon histology scored. Groups 2 and 3 were administered either control mAb or anti–IL-23p19 mAb at 100 μg/mouse weekly for 4 weeks and then killed and histopathology scored. (A) Mean ± SD of quantitative histopathology scores. (B–D) Gene expression in the colons of the 3 groups measured by real-time PCR. Pools of RNA from 5 mice per group were analyzed and gene expression compared with ubiquitin expression. Group 1, open bars; group 2, stippled bars; group 3, black bars. (E and F) Representative histopathology from mice treated with control mAb or anti–IL-23p19 mAb, respectively. This experiment was performed twice with groups of 5 mice each time. *P < .001 vs control.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

9. Figure 8
Monoclonal anti–IL-23p19 induced apoptosis of colitogenic Th17 cells. Bir14 T cells were stimulated with CBA-pulsed APCs in the presence or absence of 10 μg/mL of anti–IL-12p70 or anti–IL-23p19. IL-17 and IFN-γ production was analyzed by intracellular staining at 24 hours, and apoptosis was measured by 7-amino-actinomycin D (7-AAD) and annexin V staining at 48 hours. Data shown are representative of 3 independent experiments.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions