1. Volume 141, Issue 5, Pages 1821-1831 (November 2011)
Retinoic Acid Attenuates Ileitis by Restoring the Balance Between T-Helper 17 and T Regulatory Cells 
Colm B. Collins, Carol M. Aherne, Douglas Kominsky, Eóin N. McNamee, Matthew D.P. Lebsack, Holger Eltzschig, Paul Jedlicka, Jesús Rivera–Nieves 
Gastroenterology 
Volume 141, Issue 5, Pages (November 2011)
DOI: /j.gastro
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2. Figure 1
Proportion of CD103+ DCs decreased during late disease in TNFΔARE LP. (A) Flow cytometric analysis of CD103+ DCs in the spleen, MLNs, and LP of 4- and 20-week-old TNFΔARE mice compared with WT littermates. Mean ± SEM of n ≥3 individual mice for each genotype and time point; *P < .05. (B) Representative plots showing the gating strategy for identifying CD103+ DCs from the MLNs and LP of 20-week-old WT mice, with overlaid histograms for CD103 on DCs from a CD103-deficient mouse of matched age. (C) Representative histograms of CD103 expression on DCs from MLNs and LP of 20-week-old WT and TNFΔARE mice.
Gastroenterology  , DOI: ( /j.gastro )
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3. Figure 2
Decreased RALDH2 expression in CD103+ and CD103Neg DCs from 4- and 20-week-old TNFΔARE mice compared with WT littermates. RALDH2 mRNA expression in isolated CD103+ and CD103Neg CD11chi/MHCII+ DCs from MLNs and LP of 4- and 20-week-old WT and TNFΔARE mice. Mean ± SEM from 3 separate experiments requiring pooled organs from 4 to 11 mice/strain/experiment. *P < .05, ***P < .001.
Gastroenterology  , DOI: ( /j.gastro )
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4. Figure 3
Increased percentage of Tregs in the spleen, MLNs, and LP of 20-week-old TNFΔARE mice compared with WT littermates. (A) The percentage of CD4+/CD25+/FoxP3+ cells in the spleen, MLNs, and ileal LP of TNFΔARE mice compared with WT mice at 20 weeks of age. Mean ± SEM for n ≥ 5 individual mice/strain for each time point; *P < .05, ***P < (B) Representative dot plots showing expression of CD4+/CD25+/FoxP3+ T lymphocytes from the LP of 20-week-old TNFΔARE mice and WT littermates.
Gastroenterology  , DOI: ( /j.gastro )
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5. Figure 4
Dynamic changes in RALDH expression in IECs likely mediated by TNF overproduction and RA deficiency. (A) Quantification of RALDH3 mRNA from IEC fractions from 4- and 20-week-old TNFΔARE mice compared with WT by real-time reverse-transcription PCR. Mean ± SEM, n = 7/strain, *P < .05. (B) Expression of RALDH1, 2, and 3 mRNA transcripts in IEC fractions from 20-week-old mice, mean ± SEM, n = 7/strain. *P < .05. (C) Analysis of RALDH3 mRNA expression from MODE-K cells cultured for 48 hours treated in triplicate with 1 μmol/L RA, 10 ng/mL TNF, 1 μmol/L LE540, or 1 μmol/L AM580 stimulation or untreated for 24 hours followed by treatment with 1 μmol/L LE540 for 24 hours (NT/LE). Mean ± SEM of 4 individual experiments, *P ≤ .05, **P < .01, ***P ≤ .001.).
Gastroenterology  , DOI: ( /j.gastro )
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6. Figure 5
RA supplementation significantly attenuated chronic ileitis in vivo. (A) Inflammatory indices from ilea of 20-week-old TNFΔARE mice treated with RA intraperitoneally twice weekly for 2 weeks or vehicle were assessed as described.28 Mean ± SEM, n = 8/treatment group, *P < .05, **P < .01. (B) Representative H&E micrographs of vehicle- and RA-treated ilea of 20-week-old TNFΔARE mice (original magnification 10×, bars = 100 μm).
Gastroenterology  , DOI: ( /j.gastro )
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7. Figure 6
RA shifts regulatory and effector balance of T cells in the inflamed ileum. (A) Frequency of CD4+/CD25+/FoxP3+ Tregs and CD4+/IL-17+ Th17 cells in indicated organs isolated from vehicle- or RA-treated 20-week-old TNFΔARE mice. (B) Dot plots showing percentages of CD4+/CD25+/FoxP3+ and CD4+/IL-17+ cells from vehicle- and RA-treated mice. Representative data from 3 independent experiments (C) MCP-1, IL-1β, IL-10, and IL-17A from 24-hour cultures of unfractionated CD3/28 stimulated lymphocytes, measured using the multiplex cytokine analysis system from Quansys Biosciences. Mean ± SEM for n ≥3 mice/group. *P ≤ .05.
Gastroenterology  , DOI: ( /j.gastro )
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8. Figure 7
RA deficiency in whole intestinal tissue of inflamed mice. (A) Expression of RALDH1, 2, and 3 mRNA in ileal tissue from 20-week-old TNFΔARE mice compared with WT littermates. Mean ± SEM, n = 7/genotype, *P ≤ .05. (B) Coulometric analysis of whole tissue extracts from WT and TNFΔARE mice showed a significant reduction in RA levels in inflamed mice ilea compared with WT controls. Mean ± SEM, n = 5/genotype, *P < .05. (C) Representative scanning electron micrographs of ilea from 20-week-old WT and TNFΔARE mice (original magnification 80×).
Gastroenterology  , DOI: ( /j.gastro )
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9. Supplementary Figure 1
Gastroenterology  , DOI: ( /j.gastro )
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10. Supplementary Figure 2
Gastroenterology  , DOI: ( /j.gastro )
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11. Supplementary Figure 3
Gastroenterology  , DOI: ( /j.gastro )
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12. Supplementary Figure 4
Gastroenterology  , DOI: ( /j.gastro )
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13. Supplementary Figure 5
Gastroenterology  , DOI: ( /j.gastro )
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14. Supplementary Figure 6
Gastroenterology  , DOI: ( /j.gastro )
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15. Supplementary Figure 7
Gastroenterology  , DOI: ( /j.gastro )
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