Presentation is loading. Please wait.

Presentation is loading. Please wait.

Epithelial Overexpression of SOCS-3 in Transgenic Mice Exacerbates Wound Inflammation in the Presence of Elevated TGF-β1  Andreas Linke, Itamar Goren,

Published byKatarina Åberg Modified over 5 years ago

Similar presentations

Presentation on theme: "Epithelial Overexpression of SOCS-3 in Transgenic Mice Exacerbates Wound Inflammation in the Presence of Elevated TGF-β1  Andreas Linke, Itamar Goren,"— Presentation transcript:

1 Epithelial Overexpression of SOCS-3 in Transgenic Mice Exacerbates Wound Inflammation in the Presence of Elevated TGF-β1  Andreas Linke, Itamar Goren, Michael R. Bösl, Josef Pfeilschifter, Stefan Frank  Journal of Investigative Dermatology  Volume 130, Issue 3, Pages (March 2010) DOI: /jid Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 IL-1β and tumor necrosis factor (TNF)-α expression in disturbed wounds of tsgn-K5/SOCS-3 mice. RNase protection assays showing IL-1β (a) or TNF-α (b) mRNA expression in nonwounded skin (ctrl skin) or wound tissues at different time points after injury of transgenic mice (tsgn-K5/SOCS3) or their nontransgenic littermates (non-tsgn) as indicated (upper panels). Hybridization against glyceraldehyde-3-phosphate dehydrogenase is shown as a loading control. Quantification of the RNase protection assays (PhosphoImager PSL counts per 15μg of total wound RNA) for the 5- and 7-day time points is shown in the middle panels. **P<0.01; *P<0.05 as indicated by the brackets. Bars depict means±SEM obtained from wounds (n=12) isolated from animals (n=4) of three independent animal experiments (n=3). IL-1β- and TNF-α-specific ELISA of 5- and 7-day wound lysates isolated from transgenic (tsgn-K5/SOCS3) and nontransgenic (non-tsgn) mice as indicated (lower panels). *P<0.05 as indicated by the brackets. Bars indicate the mean±SEM of 10 wounds (n=10) of 10 animals (n=10). Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Chemokine expression in wound keratinocytes of tsgn-K5/SOCS-3 mice. RNase protection assays showing macrophage inflammatory protein (MIP)-2 (a) or MCP-1 (b) mRNA expression in nonwounded skin (ctrl skin) or wound tissues at different time points after injury from transgenic mice (tsgn-K5/SOCS3) or their nontransgenic littermates (non-tsgn) as indicated (upper panels). Hybridization against glyceraldehyde-3-phosphate dehydrogenase is shown as a loading control. Quantification of the RNase protection assays (PhosphoImager PSL counts per 15μg of total wound RNA) for the 5- and 7-day time points is shown in the middle panels. **P<0.01; *P<0.05 as indicated by the brackets. Bars depict means±SEM obtained from wounds (n=12) isolated from animals (n=4) of three independent animal experiments (n=3). MIP-2- and MCP-1-specific ELISA from 5- and 7-day wound lysates isolated from transgenic (tsgn-K5/SOCS3) and nontransgenic (non-tsgn) mice as indicated (lower panels). *P<0.05; NS, not significant as indicated by the brackets. Bars indicate the mean±SEM of 10 wounds (n=10) of 10 animals (n=10). Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Immune cell infiltration. RNase protection assay showing lysozyme M (a, upper panel) and lipocalin (d, upper panel) mRNA expression in nonwounded skin (ctrl skin) or wound tissues at different time points after injury of transgenic mice (tsgn-K5/SOCS3) or their nontransgenic littermates (non-tsgn) as indicated. Hybridization against glyceraldehyde-3-phosphate dehydrogenase is shown as a loading control. Immunoblots for F4/80 (a, lower panel) or GR-1 (d, lower panel) protein is shown below. Ponceau S staining was used to control equal loading. Lysozyme M (b, c) or lipocalin (e, f) mRNA expression during skin repair is shown for the indicated time points. *P<0.05 as indicated. Bars depict means±SEM obtained from wounds (n=12) isolated from animals (n=4) of three independent animal experiments (n=3). Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Cox-2 expression at the wound site. RNase protection assay (a, upper panels) showing Cox-2 mRNA expression in nonwounded skin (ctrl skin) or wound tissues at different time points after injury of transgenic mice (tsgn-K5/SOCS3) or their nontransgenic littermates (non-tsgn) as indicated. Hybridization against glyceraldehyde-3-phosphate dehydrogenase is shown as a loading control. Quantification of the RNase protection assays (PhosphoImager PSL counts per 15μg of total wound RNA) for the 5- and 7-day time points is shown in (b). *P<0.05 as indicated by the brackets. Bars depict means±SEM obtained from wounds (n=12) isolated from animals (n=4) of three independent animal experiments (n=3). An immunoblot for Cox-2 protein is shown (a, lower panel). Ponceau S staining was used to control equal loading. (c) Quantification of Cox-2 protein expression in 5-day and 7-day wounds as indicated. **P<0.01; *P<0.05 as indicated by the brackets. Bars depict means±SEM obtained from wounds (n=4) isolated from four animals (n=4). (d) Frozen sections of 5-day wound tissue of nontransgenic (non-tsgn) and transgenic (tsgn-K5/SOCS3) mice were incubated with an antibody directed against Cox-2 protein. Immunopositive signals are highlighted by yellow arrows. he, hyperproliferative epithelia; sc, scab. Bar=100μm. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Expression of transforming growth factor (TGF)-β1 during skin repair. (a) RNase protection assay showing TGF-β1 mRNA expression in nonwounded skin (ctrl skin) or wound tissues at different time points after injury of transgenic mice (tsgn-K5/SOCS3) or their nontransgenic littermates (non-tsgn) as indicated. Hybridization against glyceraldehyde-3-phosphate dehydrogenase is shown as a loading control. Quantification of TGF-β1 mRNA (PhosphoImager PSL counts per 15μg of total wound RNA) is shown in (b). *P<0.05 in comparison to nontransgenic littermates. Bars depict means±SEM obtained from wounds (n=12) isolated from animals (n=4) of three independent animal experiments (n=3). (c) TGF-β1-specific ELISA of 5-day and 7-day wound lysates and nonwounded skin (ctrl) isolated from transgenic (tsgn-K5/SOCS3) and nontransgenic (non-tsgn) mice as indicated. **P<0.01; *P<0.05 as indicated by the brackets. Bars depict means±SEM obtained from wounds (n=12) isolated from animals (n=6). Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 SOCS-3-overexpressing keratinocytes are responsive to inflammatory cytokines and transforming growth factor (TGF)-β1. Confluent primary keratinocytes of transgenic mice (tsgn-K5/SOCS3) or their nontransgenic littermates (non-tsgn) were assessed for the release of MCP-1 (a), macrophage inflammatory protein (MIP)-2 (b), and nitrite (c) in the presence or absence of cytokines (40ngml−1 IL-1β, 50ngml−1 tumor necrosis factor-α, 20ngml−1 IFN-γ), TGF-β1 (5ngml−1) or anti-TGF-β antibody (10μgml−1) after 24hours as indicated. **P<0.01; *P<0.05 as indicated by the brackets. Bars depict means±SEM obtained from three independent cell culture experiments (n=3). Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

8 Figure 7 Application of an anti–transforming growth factor(TGF)-β antibody improves wound closure in tsgn-K5/SOCS-3 mice. (a) In situ wound phenotypes of acute 5-day wounds of tsgn-K5/SOCS-3 mice (right panels) and their nontransgenic littermates (left panels) upon control-IgG or anti-TGF-β treatment. (b) Hematoxylin-stained frozen sections of 5-day wounds isolated from tsgn-K5/SOCS-3 mice upon control-IgG or anti-TGF-β treatment. The distance between the hyperproliferative epithelia at the wound margins is indicated by black arrows. The epithelial boundaries are highlighted by a red line. gt, granulation tissue; he, hyperproliferative epithelium; sc, scab. Bars=(a) 5mm, (b) 600μm. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

9 Figure 8 Neutralization of transforming growth factor (TGF)-β in wounds of tsgn-K5/SOCS-3 mice. (a) RNase protection assays were performed to determine IL-1β (a), MIP-2 (b), and Cox-2 (c) mRNA expression in 5-day wound tissue of transgenic mice (tsgn-K5/SOCS3) or their nontransgenic littermates (non-tsgn) upon control-IgG or anti-TGF-β treatment. The quantification of assays (PhosphoImager PSL counts per 15μg of total wound RNA) is shown. **P<0.01; *P<0.05, NS, not significant as indicated by the brackets. Data represent means±SEM obtained from eight wounds isolated from four animals (n=8). (d) TGF-β-sensitive mink lung epithelial reporter cells were stimulated with 5-day wound lysates of transgenic mice (tsgn-K5/SOCS3) or nontransgenic littermates (non-tsgn) and recombinant TGF-β1 (800pgml−1) in the presence or absence of a TGF-β-neutralizing antibody for 20hours as indicated. TGF-β bioactivity is visualized by relative luminescence units (RLUs) derived from luciferase enzymatic activity. Cells cultured on serum-free media served as a control (ctrl). **P<0.01 as indicated by the brackets. Bars depict means±SEM obtained from independent cell culture experiments (n=8). Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

Download ppt "Epithelial Overexpression of SOCS-3 in Transgenic Mice Exacerbates Wound Inflammation in the Presence of Elevated TGF-β1  Andreas Linke, Itamar Goren,"

Similar presentations

Volume 39, Issue 5, Pages (November 2013)

Volume 39, Issue 5, Pages (November 2013)
The Suppressor of Cytokine Signaling-3 Is Upregulated in Impaired Skin Repair: Implications for Keratinocyte Proliferation  Itamar Goren, Andreas Linke,

The Suppressor of Cytokine Signaling-3 Is Upregulated in Impaired Skin Repair: Implications for Keratinocyte Proliferation  Itamar Goren, Andreas Linke,
Interleukin-22 Promotes Wound Repair in Diabetes by Improving Keratinocyte Pro- Healing Functions  Simona Avitabile, Teresa Odorisio, Stefania Madonna,

Interleukin-22 Promotes Wound Repair in Diabetes by Improving Keratinocyte Pro- Healing Functions  Simona Avitabile, Teresa Odorisio, Stefania Madonna,
Loss of Extracellular Superoxide Dismutase Induces Severe IL-23-Mediated Skin Inflammation in Mice  Yun Sang Lee, In-Su Cheon, Byung-Hak Kim, Myung-Ja.

Loss of Extracellular Superoxide Dismutase Induces Severe IL-23-Mediated Skin Inflammation in Mice  Yun Sang Lee, In-Su Cheon, Byung-Hak Kim, Myung-Ja.
Cdc42 Inhibits ERK-Mediated Collagenase-1 (MMP-1) Expression in Collagen-Activated Human Keratinocytes  Maryam G. Rohani, Brian K. Pilcher, Peter Chen,

Cdc42 Inhibits ERK-Mediated Collagenase-1 (MMP-1) Expression in Collagen-Activated Human Keratinocytes  Maryam G. Rohani, Brian K. Pilcher, Peter Chen,
Characterization of Spink6 in Mouse Skin: The Conserved Inhibitor of Kallikrein-Related Peptidases Is Reduced by Barrier Injury  Jan Fischer, Zhihong.

Characterization of Spink6 in Mouse Skin: The Conserved Inhibitor of Kallikrein-Related Peptidases Is Reduced by Barrier Injury  Jan Fischer, Zhihong.
Birgit Stallmeyer, Heiko Kämpfer, Josef Pfeilschifter, Stefan Frank 

Birgit Stallmeyer, Heiko Kämpfer, Josef Pfeilschifter, Stefan Frank 
Counterregulation of Interleukin-18 mRNA and Protein Expression During Cutaneous Wound Repair in Mice  Heiko Kämpfer, Heiko Mühl, Josef Pfeilschifter,

Counterregulation of Interleukin-18 mRNA and Protein Expression During Cutaneous Wound Repair in Mice  Heiko Kämpfer, Heiko Mühl, Josef Pfeilschifter,
Type I IL-1 Receptor Mediates IL-1 and Intracellular IL-1 Receptor Antagonist Effects in Skin Inflammation  Gaby Palmer, Dominique Talabot-Ayer, Gürkan.

Type I IL-1 Receptor Mediates IL-1 and Intracellular IL-1 Receptor Antagonist Effects in Skin Inflammation  Gaby Palmer, Dominique Talabot-Ayer, Gürkan.
Inhibition of UVB-Induced Skin Tumor Development by Drinking Green Tea Polyphenols Is Mediated Through DNA Repair and Subsequent Inhibition of Inflammation 

Inhibition of UVB-Induced Skin Tumor Development by Drinking Green Tea Polyphenols Is Mediated Through DNA Repair and Subsequent Inhibition of Inflammation 
IL-17A RNA Aptamer: Possible Therapeutic Potential in Some Cells, More than We Bargained for in Others?  Rosella Doble, Michael F. McDermott, Özlem Cesur,

IL-17A RNA Aptamer: Possible Therapeutic Potential in Some Cells, More than We Bargained for in Others?  Rosella Doble, Michael F. McDermott, Özlem Cesur,
Impaired Wound Repair in Adult Endoglin Heterozygous Mice Associated with Lower NO Bioavailability  Eduardo Pérez-Gómez, Mirjana Jerkic, Marta Prieto,

Impaired Wound Repair in Adult Endoglin Heterozygous Mice Associated with Lower NO Bioavailability  Eduardo Pérez-Gómez, Mirjana Jerkic, Marta Prieto,
Decreased Expression of Caveolin-1 Contributes to the Pathogenesis of Psoriasiform Dermatitis in Mice  Yukie Yamaguchi, Yuko Watanabe, Tomoya Watanabe,

Decreased Expression of Caveolin-1 Contributes to the Pathogenesis of Psoriasiform Dermatitis in Mice  Yukie Yamaguchi, Yuko Watanabe, Tomoya Watanabe,
Stromal Fibroblast–Specific Expression of ADAM-9 Modulates Proliferation and Apoptosis in Melanoma Cells In Vitro and In Vivo  Anna N. Abety, Jay W. Fox,

Stromal Fibroblast–Specific Expression of ADAM-9 Modulates Proliferation and Apoptosis in Melanoma Cells In Vitro and In Vivo  Anna N. Abety, Jay W. Fox,
Accelerated Wound Repair in ADAM-9 Knockout Animals

Accelerated Wound Repair in ADAM-9 Knockout Animals
Large and Sustained Induction of Chemokines during Impaired Wound Healing in the Genetically Diabetic Mouse: Prolonged Persistence of Neutrophils and.

Large and Sustained Induction of Chemokines during Impaired Wound Healing in the Genetically Diabetic Mouse: Prolonged Persistence of Neutrophils and.
Oncostatin M Expression Is Functionally Connected to Neutrophils in the Early Inflammatory Phase of Skin Repair: Implications for Normal and Diabetes-Impaired.

Oncostatin M Expression Is Functionally Connected to Neutrophils in the Early Inflammatory Phase of Skin Repair: Implications for Normal and Diabetes-Impaired.
IL-27 Activates Th1-Mediated Responses in Imiquimod-Induced Psoriasis-Like Skin Lesions  Sayaka Shibata, Yayoi Tada, Yoshihide Asano, Koichi Yanaba, Makoto.

IL-27 Activates Th1-Mediated Responses in Imiquimod-Induced Psoriasis-Like Skin Lesions  Sayaka Shibata, Yayoi Tada, Yoshihide Asano, Koichi Yanaba, Makoto.
Volume 39, Issue 5, Pages (November 2013)

Volume 39, Issue 5, Pages (November 2013)
Manuela Schmidt, Danny Gutknecht, Jan C

Manuela Schmidt, Danny Gutknecht, Jan C

Similar presentations

Ads by Google