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Science 329 (5993): 849-853

Copyright © 2010 by the American Association for the Advancement of Science

Activation of β-Catenin in Dendritic Cells Regulates Immunity Versus Tolerance in the Intestine

Santhakumar Manicassamy1, Boris Reizis2, Rajesh Ravindran1, Helder Nakaya1, Rosa Maria Salazar-Gonzalez1,3, Yi-chong Wang1, and Bali Pulendran1,4,*

1 Emory Vaccine Center, and Yerkes National Primate Research Center, 954 Gatewood Road, Atlanta, GA 30329, USA.
2 Department of Microbiology, Columbia University Medical Center, New York, NY 10032, USA.
3 Molecular Immunology Division, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229–3039, USA.
4 Department of Pathology, Emory University, 1364 Clifton Road NE, Atlanta, GA 30322, USA.


Figure 1
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Fig. 1. β-catenin signaling is constitutively active in intestinal APCs and regulates the induction of Treg cells and effector T cells. (A) Expression of β-galactosidase (representing β-catenin activity) by intestinal DCs and macrophages from SI-LP and LI-LP of TCF-reporter mice. (B and C) Fluorescence-activated cell sorting (FACS) plots representing percentages of CD4+ T cells positive for FoxP3 (Treg cells), IL-17 (TH17), and IFN-{gamma} (TH1) isolated from SI-LP, LI-LP, ceacum, and spleen of β-catfl/fl and β-catDC–/– mice. (D and E) Intracellular expression of FoxP3, IL-17, and IFN-{gamma} in naïve CD4+OT-II T cells stimulated to differentiate in vitro by intestinal APCs (CD11c+CD11b and CD11c+CD11b+ DCs and CD11cCD11b+ macrophages) isolated from β-catfl/fl or β-catDC–/– mice, in the presence of TGF-β (1 ng/ml). Numbers in FACS plots represent percentage of cells positive for the indicated protein. Data are from one experiment representative of three.

 

Figure 2
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Fig. 2. β-catenin signaling in intestinal DCs promotes the expression of Raldh and suppresses the expression of proinflammatory cytokines. (A) Expression of Aldh1a1 and Aldh1a2 mRNA in CD11c+CD11b and CD11c+CD11b+ DCs and CD11cCD11b+ macrophages isolated from SI-LP of β-catfl/fl or β-catDC–/– mice. (B) Expression of Raldh protein by CD11c+CD11b and CD11c+CD11b+ DCs and CD11c CD11b+ macrophages isolated from SI-LP of β-catfl/fl or β-catDC–/– mice, as assessed by intracellular staining and flow cytometry. (C) Expression of Il-10, Tgf-β1, Il-23a, and Il-6 mRNAs in CD11c+CD11b and CD11c+CD11b+ DCs and CD11cCD11b+ macrophages isolated from SI-LP of β-catfl/fl or β-catDC–/– mice. (D) Cytokine concentrations in the supernatants of CD11c+CD11b and CD11c+CD11b+ DCs and CD11cCD11b+ macrophages isolated from SI-LP of β-catfl/fl or β-catDC–/– mice after 24 hours. *P < 0.05; **P < 0.005. Error bars indicate mean ± SEM. Data are representative of three experiments.

 

Figure 3
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Fig. 3. β-catenin activation in intestinal DCs is independent of commensals and induced by Wnt-signaling. (A and B) FACS plots representing the percentage of CD4+ T cells that express (A) FoxP3, IL-17, and (B) IFN-{gamma} isolated from SI-LP of β-catfl/fl or β-catDC–/– mice treated with (Atx) or without (None) antibiotics. (C) FACS plots showing the intracellular expression levels of β-galactosidase or β-catenin protein in CD11c+CD11b and CD11c+CD11b+ DCs and CD11cCD11b+ macrophages isolated from SI-LP of TCF-reporter mice treated with or without antibiotics. (D and E) mRNA expression of (D) Fzd receptors and (E) Wnt ligands in CD11c+CD11b and CD11c+CD11b+ DCs and CD11cCD11b+ macrophages isolated from SI-LP of β-catfl/fl mice. (F) mRNA expression levels of Wnt–β-catenin target genes Wisp1, Wisp2, and Axin1 in CD11c+CD11b and CD11c+CD11b+ DCs and CD11cCD11b+ macrophages isolated from SI-LP of β-catfl/fl mice. Error bars indicate mean ± SEM. Data are from one experiment representative of three.

 

Figure 4
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Fig. 4. β-catenin signaling in LP-DCs regulates intestinal homeostasis. (A) Percent body weight of β-catfl/fl or β-catDC–/– mice treated with 2% DSS for 7 days. Error bars indicate mean ± SD. (B) FACS plot representing percentage of CD4+ cells positive for IL-17 and IFN-{gamma} isolated on day 8, from Ceacum and LI-LP of β-catfl/fl or β-catDC–/– mice treated with DSS for 6 days. Data are from one experiment representative of three. (C) Histopathological changes in colon tissue from β-catfl/fl or β-catDC–/– mice treated with or without 2% DSS treatment for 7 days. Areas of interest are infiltrations, edema (yellow arrows), globlet cells (black arrows), and basement membrane (green arrows). Images are 10x original magnification.

 


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