Presentation is loading. Please wait.

Presentation is loading. Please wait.

Conditional PDK1 Ablation Promotes Epidermal and T-Cell-Mediated Dysfunctions Leading to Inflammatory Skin Disease  Minjun Yu, David M. Owens, Sankar.

Published byMeredith Short Modified over 5 years ago

Similar presentations

Presentation on theme: "Conditional PDK1 Ablation Promotes Epidermal and T-Cell-Mediated Dysfunctions Leading to Inflammatory Skin Disease  Minjun Yu, David M. Owens, Sankar."— Presentation transcript:

1 Conditional PDK1 Ablation Promotes Epidermal and T-Cell-Mediated Dysfunctions Leading to Inflammatory Skin Disease  Minjun Yu, David M. Owens, Sankar Ghosh, Donna L. Farber  Journal of Investigative Dermatology  Volume 135, Issue 11, Pages (November 2015) DOI: /jid Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Conditional ablation of phosphoinositide-dependent kinase-1 (PDK1) in OX40-expressing cells results in diffuse dermatitis, fibrosis, and Th2 polarization. (a) Macroscopic view of PDK1-CKO and PDK1-CHET mice (left), cervical lymph nodes (upper right), and spleen (lower right) from each strain. (b) Survival curve of PDK1-CHET (n=20) and PDK1-CKO (n=15) mice. (c) Skin sections from PDK1-CKO and PDK1-CHET mice showing hematoxylin and eosin staining (upper row) and Masson Trichrome staining (lower row). Bars=100 μm (*Indicates epidermal hyperplasia). (d) Flow cytometric analysis of CD4 T cells from peripheral lymph nodes of PDK-CKO and PDK1-CHET mice (7–8 weeks old). Left: forward scatter versus side scatter; middle: yellow fluorescent protein (YFP) expression by CD4 T cells (shown as mean±s.e.m., n=6); right: CD44 versus CD62L expression of CD4+YFP+ cells. (e) IL-4 production from PDK1-CKO CD4 T cells expressed as percentage of YFP+IL-4+ in each quadrant drawn based on control unstimulated T cells. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Phosphoinositide-dependent kinase-1 (PDK1)-deficient T cells promote skin pathology and mild disease in RAG2-/- hosts. (a) Macroscopic view of RAG2-/- recipients 13 weeks posttransfer of 5 × 106 CD3+ T cells from PDK1-CKO (“RAG2+CKO”, left) or PDK1-CHET (“RAG2+CHET”, right) mice. (b) Disease score (see Materials and Methods) of RAG2+CHET (n=5) or RAG2+CKO (n=8) recipients of T cells 12 weeks posttransfer compared with the disease score of 8-week-old PDK1-CKO parental mice. (c) Skin sections of RAG2+CKO and RAG2+CHET recipient mice stained with hematoxylin and eosin (upper) or Masson Trichrome (lower). Bars=100 μm. (d) Immunofluorescence of CD4 (first column) and yellow fluorescent protein (YFP; second column) expression together with DAPI (4,6-diamidino-2-phenylindole) nuclear staining (third column) in skin sections from RAG2+CKO (upper row) and RAG2+CHET (lower row) mice. Bars=25 μm. D, dermis; ED, epidermis. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 OX40-Cre-mediated phosphoinositide-dependent kinase-1 (PDK1) ablation occurs in epidermal cells of PDK1-CKO mice. (a) Left, Immunofluorescence of CD4 and yellow fluorescent protein (YFP) expression with DAPI (4,6-diamidino-2-phenylindole) nuclear stain in the skin of PDK1-CKO mice and PDK1-CHET mice from six experiments. Bars=25 μm. Right, PDK1 transcript expression (mean±s.e.m., n=3) in YFP+ epidermal cells measured by quantitative PCR (qPCR), calculated relative to levels in PDK1-CHET/YFP+ normalized to 1. (b) OX40 expression by qPCR in YFP+ cells sorted from PDK1-CKO, -CHET, and OX40-/-/ROSAYFP mice at 3 and/or 7 weeks of age, expressed relative to levels in PDK1-CHET cells (normalized to 1.0). (c) Ki67 expression in the skin of PDK1-CKO and PDK1-CHET mice (7–8 weeks old). Bars=50 μm. (d) Immunofluorescence of YFP (green), cytokeratin 14 (red), and DAPI (third column) in the skin of PDK1-CKO mice at the indicated ages. D, dermis; ED, epidermis. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Lymphocyte-deficient PDK1-CKO mice spontaneously develop skin pathology and disease symptoms. (a) Macroscopic view of PDK1-CKO × RAG2+/- (CKOXRAG2+/-) and PDK1-CKO × RAG2-/- (CKO × RAG2-/-) mice, with corresponding hematoxylin and eosin–stained skin sections obtained at 8 weeks of age. Bars=100 μm. (b) Disease score of CKO × RAG2+/- (n=6) and CKO × RAG2-/- mice (n=8). (c) Survival curve of CKO × RAG2-/- (n=12) mice superimposed on the survival curve of PDK1-CHET (n=20) and PDK1-CKO (n=15) mice. (d) Skin sections stained with Ki67 (red), yellow fluorescent protein (YFP; yellow), and DAPI (4,6-diamidino-2-phenylindole; blue) as indicated. Bars=50 μm. D, dermis; ED, epidermis. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Phosphoinositide-dependent kinase-1 (PDK1) ablation in skin impairs keratinocyte differentiation and increases inflammation. (a) Left, Immunofluorescence staining of keratin-10 (upper) and loricrin (lower) in skin sections from PDK1-CKO and PDK1-CHET mice. Bars=25 μm. Right, quantitative PCR (qPCR) of loricrin expression (average of triplicates) in sorted YFP+ epidermal cells. (b) Immunofluorescence of keratin-10 (upper) and loricrin (lower) staining in skin sections from CKO × RAG2-/- and CHET × RAG2-/- mice. Bars=50 μm. (c) Left, Immunofluorescence of yellow fluorescent protein (YFP) and thymic stromal lymphopoietin (TSLP) expression in the skin of PDK1-CKO mice and PDK1-CHET mice (7 weeks old). Bar=25 μm. (d) TSLP transcript expression level (qPCR, average of triplicates) in sorted YFP+ epidermal cells of PDK1-CKO and -CHET mice (upper) and YFP+α6-integrin+ cells of CKO × RAG2-/- and CHET × RAG2-/- skin (lower). (e) Serum TSLP level from 6–8-week-old mice as indicated. *** represents P< NS, not significant. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 Phosphoinositide-dependent kinase-1 (PDK1) ablation in keratinocytes is sufficient for inducing skin infiltration and T helper type 2 activation. T cells from PDK1-CHET mice were transferred to CHET × RAG2-/- or CKO × RAG2-/- recipients (see Materials and Methods), resulting in CHET × RAG2-/-+CHET or CKO × RAG2-/-+CHET mice, respectively. (a) Disease score of recipient strains 2 weeks posttransfer for CHET × RAG2-/- (n=4) and CKO × RAG2-/- (n=5) mice, compared with disease score of age-matched CKOXRAG2-/- mice (n=5) without transferred T cells. (b) Skin sections (hematoxylin and eosin stained) from CHET × RAG2-/-+CHET and CKO × RAG2-/-+CHET mice 2 weeks posttransfer. Bars: I–II, 100 μm; III–IV, 50 μm. (c) Immunofluorescence of CD4 and yellow fluorescent protein (YFP) expression in the skin of the indicated strains. Bars=50 μm. (d) IL-4 production by T cells isolated from CHET × RAG2-/-+CHET or CKO × RAG2-/-+CHET mice expressed as mean frequency±s.e.m. of CD4+YFP+ IL-4+ cells after 5-h stimulation. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

Download ppt "Conditional PDK1 Ablation Promotes Epidermal and T-Cell-Mediated Dysfunctions Leading to Inflammatory Skin Disease  Minjun Yu, David M. Owens, Sankar."

Similar presentations

Volume 39, Issue 5, Pages (November 2013)

Volume 39, Issue 5, Pages (November 2013)
IL-17A as an Inducer for Th2 Immune Responses in Murine Atopic Dermatitis Models  Saeko Nakajima, Akihiko Kitoh, Gyohei Egawa, Yohei Natsuaki, Satoshi.

IL-17A as an Inducer for Th2 Immune Responses in Murine Atopic Dermatitis Models  Saeko Nakajima, Akihiko Kitoh, Gyohei Egawa, Yohei Natsuaki, Satoshi.
Psoriatic Inflammation Facilitates the Onset of Arthritis in a Mouse Model  Mayuko Yamamoto, Kimiko Nakajima, Mikiro Takaishi, Shun Kitaba, Yasuhiro Magata,

Psoriatic Inflammation Facilitates the Onset of Arthritis in a Mouse Model  Mayuko Yamamoto, Kimiko Nakajima, Mikiro Takaishi, Shun Kitaba, Yasuhiro Magata,
The Tumor Necrosis Factor Superfamily Molecule LIGHT Promotes Keratinocyte Activity and Skin Fibrosis  Rana Herro, Ricardo Da S. Antunes, Amelia R. Aguilera,

The Tumor Necrosis Factor Superfamily Molecule LIGHT Promotes Keratinocyte Activity and Skin Fibrosis  Rana Herro, Ricardo Da S. Antunes, Amelia R. Aguilera,
Autoantibodies in Scurfy Mice and IPEX Patients Recognize Keratin 14

Autoantibodies in Scurfy Mice and IPEX Patients Recognize Keratin 14
Transcriptome Analysis of Psoriasis in a Large Case–Control Sample: RNA-Seq Provides Insights into Disease Mechanisms  Bingshan Li, Lam C. Tsoi, William.

Transcriptome Analysis of Psoriasis in a Large Case–Control Sample: RNA-Seq Provides Insights into Disease Mechanisms  Bingshan Li, Lam C. Tsoi, William.
Gregory D. Rak, Lisa C. Osborne, Mark C. Siracusa, Brian S

Gregory D. Rak, Lisa C. Osborne, Mark C. Siracusa, Brian S
UVB-Induced Skin Inflammation and Cutaneous Tissue Injury Is Dependent on the MHC Class I–Like Protein, CD1d  Stephan Ryser, Marlène Schuppli, Beatrice.

UVB-Induced Skin Inflammation and Cutaneous Tissue Injury Is Dependent on the MHC Class I–Like Protein, CD1d  Stephan Ryser, Marlène Schuppli, Beatrice.
Mesenchymal Stem Cells (MSCs) Attenuate Cutaneous Sclerodermatous Graft-Versus- Host Disease (Scl-GVHD) through Inhibition of Immune Cell Infiltration.

Mesenchymal Stem Cells (MSCs) Attenuate Cutaneous Sclerodermatous Graft-Versus- Host Disease (Scl-GVHD) through Inhibition of Immune Cell Infiltration.
Pharmacologically Antagonizing the CXCR4-CXCL12 Chemokine Pathway with AMD3100 Inhibits Sunlight-Induced Skin Cancer  Seri N.E. Sarchio, Richard A. Scolyer,

Pharmacologically Antagonizing the CXCR4-CXCL12 Chemokine Pathway with AMD3100 Inhibits Sunlight-Induced Skin Cancer  Seri N.E. Sarchio, Richard A. Scolyer,
Topical ROR Inverse Agonists Suppress Inflammation in Mouse Models of Atopic Dermatitis and Acute Irritant Dermatitis  Jun Dai, Min-Kyung Choo, Jin Mo.

Topical ROR Inverse Agonists Suppress Inflammation in Mouse Models of Atopic Dermatitis and Acute Irritant Dermatitis  Jun Dai, Min-Kyung Choo, Jin Mo.
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.
Alexandra Charruyer, Chantal O. Barland, Lili Yue, Heike B

Alexandra Charruyer, Chantal O. Barland, Lili Yue, Heike B
Regulation and Function of the Caspase-1 in an Inflammatory Microenvironment  Dai-Jen Lee, Fei Du, Shih-Wei Chen, Manando Nakasaki, Isha Rana, Vincent.

Regulation and Function of the Caspase-1 in an Inflammatory Microenvironment  Dai-Jen Lee, Fei Du, Shih-Wei Chen, Manando Nakasaki, Isha Rana, Vincent.
Andrew L. Croxford, Susanne Karbach, Florian C

Andrew L. Croxford, Susanne Karbach, Florian C
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,
Shilpak Chatterjee, Jonathan M. Eby, Amir A

Shilpak Chatterjee, Jonathan M. Eby, Amir A
Tumor Necrosis Factor-α-Activated Human Adipose Tissue–Derived Mesenchymal Stem Cells Accelerate Cutaneous Wound Healing through Paracrine Mechanisms 

Tumor Necrosis Factor-α-Activated Human Adipose Tissue–Derived Mesenchymal Stem Cells Accelerate Cutaneous Wound Healing through Paracrine Mechanisms 
Preclinical Studies of a Specific PPARγ Modulator in the Control of Skin Inflammation  Arianna Mastrofrancesco, Daniela Kovacs, Massimiliano Sarra, Emanuela.

Preclinical Studies of a Specific PPARγ Modulator in the Control of Skin Inflammation  Arianna Mastrofrancesco, Daniela Kovacs, Massimiliano Sarra, Emanuela.
Induction of Thymic Stromal Lymphopoietin Expression in Keratinocytes Is Necessary for Generating an Atopic Dermatitis upon Application of the Active.

Induction of Thymic Stromal Lymphopoietin Expression in Keratinocytes Is Necessary for Generating an Atopic Dermatitis upon Application of the Active.

Similar presentations

Ads by Google