Presentation is loading. Please wait.

Presentation is loading. Please wait.

UVB and Proinflammatory Cytokines Synergistically Activate TNF-α Production in Keratinocytes through Enhanced Gene Transcription  Muhammad M. Bashir,

Published byEsa Penttilä Modified over 5 years ago

Similar presentations

Presentation on theme: "UVB and Proinflammatory Cytokines Synergistically Activate TNF-α Production in Keratinocytes through Enhanced Gene Transcription  Muhammad M. Bashir,"— Presentation transcript:

1 UVB and Proinflammatory Cytokines Synergistically Activate TNF-α Production in Keratinocytes through Enhanced Gene Transcription  Muhammad M. Bashir, Meena R. Sharma, Victoria P. Werth  Journal of Investigative Dermatology  Volume 129, Issue 4, Pages (April 2009) DOI: /jid Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 IL-1α synergistically increases TNF-α mRNA induction by UVB-irradiated neonatal KCs. Cells were treated with sham, sham+IL-1α (10ngml−1), UVB (30mJcm−2), and UVB+IL-1α. RNA samples were collected at the indicated times and analyzed by real-time PCR. TNF-α mRNA cycle numbers were normalized to cyclophilin A (PPIA). Expression levels of TNF-α mRNA are indicated as “fold change” compared to control. UVB+IL-1α-treated cells showed synergistically increased TNF-α mRNA as compared to UVB or IL-1α alone (Figure 1a and b). The time-course study showed that there was no biphasic induction of TNF-α mRNA in keratinocytes up to 72hours. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Anti-TNF-α antibody has no effect on TNF-α mRNA synthesis in UVB-irradiated keratinocytes in the presence of IL-1α. Conditions included cells treated with sham, sham+IL-1α, UVB, and UVB+IL-1α. In the test group, UVB+ IL-1α-treated cells were also incubated with anti-TNF-α antibody (5, 10, 100ngml−1). RNA samples were collected after 3hours and analyzed by real-time PCR. Results were normalized to PPIA mRNA and compared to control groups. No significant difference in expression of TNF-α mRNA is observed between UVB+ IL-1α and UVB+ IL-1α+anti-TNF-α-treated keratinocytes. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 IFN-α2b has no effect on TNF-α mRNA synthesis in sham- or UVB-irradiated keratinocytes in the presence or absence of IL-1α. Sham- or UVB-irradiated keratinocytes were treated with IFN-α2b (2,000Uml−1) or IL-1α (10ngml−1) alone or in combination with IFN-α2b+IL-1α. RNA sample were collected after 6hours and analyzed by real-time PCR. Results were normalized to PPIA mRNA and compared to control groups. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Exogenous rTNF-α increases TNF-α mRNA induction in UVB-irradiated neonatal keratinocytes. Cells were treated with sham, sham+rTNF-α (5ngml−1), UVB, and UVB+rTNF-α. RNA sample were collected 3hours after irradiation and analyzed by real-time PCR. Results were normalized to PPIA (housekeeping gene control). Expression levels of TNF-α mRNA are indicated as “fold change” compared to control. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Synergistic increase of TNF-α protein in keratinocyte medium corresponds to TNF-α mRNA increase due to UVB and IL-1α response. Cells were treated with sham, sham+IL-1α, UVB, and UVB+IL-1α for various time points. Supernatant samples were collected from sham, sham+IL-1α, UVB, and UVB+IL-1α-treated cells at the indicated time points. An equal amount of protein was used for TNF-α ELISA. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 No significant changes were observed in the membrane bound TNF-α protein after UVB and IL-1α treatment. Cells were treated with sham, sham+IL-1α, UVB, and UVB+IL-1α for various time points. Membrane fractions were eluted and an equal amount of protein was used for TNF-α ELISA. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

8 Figure 7 UVB irradiation and IL-1α enhances TNF-α gene transcription. Nuclei were isolated 3hours after sham- or UVB-irradiation and stimulated with or without IL-1α. Nuclear run-off assay was performed to examine TNF-α and 18S (control) transcription. Detectable bands were assessed by densitometric analysis. UVB irradiation induced TNF-α gene transcription five-fold. One of four experiments is shown illustrating 11-fold induction of UVB+ IL-1α cells when compared with sham irradiated cells. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

9 Figure 8 TNF-α mRNA stability is not induced after UVB irradiation and IL-1α stimulation. Keratinocytes were UVB- or sham-irradiated and stimulated with IL-1α for 3hours. Actinomycin D (10μgml−1) was added after 3hours. Total RNA was extracted at different time points and TNF-α mRNA levels were determined by northern analysis and real-time PCR. The half-life was calculated in UVB, and UVB+IL-1α-treated cells. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

Download ppt "UVB and Proinflammatory Cytokines Synergistically Activate TNF-α Production in Keratinocytes through Enhanced Gene Transcription  Muhammad M. Bashir,"

Similar presentations

Glucocorticoids Augment the Chemically Induced Production and Gene Expression of Interleukin-1α through NF-κB and AP-1 Activation in Murine Epidermal.

Glucocorticoids Augment the Chemically Induced Production and Gene Expression of Interleukin-1α through NF-κB and AP-1 Activation in Murine Epidermal.
IL-18 Downregulates Collagen Production in Human Dermal Fibroblasts via the ERK Pathway  Hee Jung Kim, Seok Bean Song, Jung Min Choi, Kyung Moon Kim,

IL-18 Downregulates Collagen Production in Human Dermal Fibroblasts via the ERK Pathway  Hee Jung Kim, Seok Bean Song, Jung Min Choi, Kyung Moon Kim,
Anti-Apoptotic NF-κB and “Gain of Function” mutp53 in Concert Act Pro-Apoptotic in Response to UVB+IL-1 via Enhanced TNF Production  Ines Müller, Stefan.

Anti-Apoptotic NF-κB and “Gain of Function” mutp53 in Concert Act Pro-Apoptotic in Response to UVB+IL-1 via Enhanced TNF Production  Ines Müller, Stefan.
Oliver Schnorr, Christoph V. Suschek, Victoria Kolb-Bachofen 

Oliver Schnorr, Christoph V. Suschek, Victoria Kolb-Bachofen 
CD271 on Melanoma Cell Is an IFN-γ-Inducible Immunosuppressive Factor that Mediates Downregulation of Melanoma Antigens  Junpei Furuta, Takashi Inozume,

CD271 on Melanoma Cell Is an IFN-γ-Inducible Immunosuppressive Factor that Mediates Downregulation of Melanoma Antigens  Junpei Furuta, Takashi Inozume,
Differential Regulation by IL-1β and EGF of Expression of Three Different Hyaluronan Synthases in Oral Mucosal Epithelial Cells and Fibroblasts and Dermal.

Differential Regulation by IL-1β and EGF of Expression of Three Different Hyaluronan Synthases in Oral Mucosal Epithelial Cells and Fibroblasts and Dermal.
Yeonsue Jang, Sang H. Jeong, Yoon-Hee Park, Hyun C

Yeonsue Jang, Sang H. Jeong, Yoon-Hee Park, Hyun C
Interferon-Gamma Enhances TLR3 Expression and Anti-Viral Activity in Keratinocytes  A.i. Kajita, Shin Morizane, Tetsuya Takiguchi, Takenobu Yamamoto, Masao.

Interferon-Gamma Enhances TLR3 Expression and Anti-Viral Activity in Keratinocytes  A.i. Kajita, Shin Morizane, Tetsuya Takiguchi, Takenobu Yamamoto, Masao.
Volume 71, Issue 6, Pages (March 2007)

Volume 71, Issue 6, Pages (March 2007)
Staphylococcus aureus Stimulates Neutrophil Targeting Chemokine Expression in Keratinocytes through an Autocrine IL-1α Signaling Loop  Florina Olaru,

Staphylococcus aureus Stimulates Neutrophil Targeting Chemokine Expression in Keratinocytes through an Autocrine IL-1α Signaling Loop  Florina Olaru,
Interferon-γ Protects from Staphylococcal Alpha Toxin-Induced Keratinocyte Death through Apolipoprotein L1  Anne M. Brauweiler, Elena Goleva, Donald Y.M.

Interferon-γ Protects from Staphylococcal Alpha Toxin-Induced Keratinocyte Death through Apolipoprotein L1  Anne M. Brauweiler, Elena Goleva, Donald Y.M.
IL-13-Stimulated Human Keratinocytes Preferentially Attract CD4+CCR4+ T cells: Possible Role in Atopic Dermatitis  Rahul Purwar, Thomas Werfel, Miriam.

IL-13-Stimulated Human Keratinocytes Preferentially Attract CD4+CCR4+ T cells: Possible Role in Atopic Dermatitis  Rahul Purwar, Thomas Werfel, Miriam.
Cytokine Effects Induced by the Human Autoallergen α-NAC

Cytokine Effects Induced by the Human Autoallergen α-NAC
Interleukin-17 and Interferon-γ Synergize in the Enhancement of Proinflammatory Cytokine Production by Human Keratinocytes  Marcel B.M. Teunissen, Jan.

Interleukin-17 and Interferon-γ Synergize in the Enhancement of Proinflammatory Cytokine Production by Human Keratinocytes  Marcel B.M. Teunissen, Jan.
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.
Volume 54, Issue 1, Pages (July 1998)

Volume 54, Issue 1, Pages (July 1998)
Streptococcus Induces Circulating CLA+ Memory T-Cell-Dependent Epidermal Cell Activation in Psoriasis  Marta Ferran, Ana B. Galván, Catalina Rincón, Ester.

Streptococcus Induces Circulating CLA+ Memory T-Cell-Dependent Epidermal Cell Activation in Psoriasis  Marta Ferran, Ana B. Galván, Catalina Rincón, Ester.
EGFR Regulates the Expression of Keratinocyte-Derived Granulocyte/Macrophage Colony-Stimulating Factor In Vitro and In Vivo  Francesca Mascia, Christophe.

EGFR Regulates the Expression of Keratinocyte-Derived Granulocyte/Macrophage Colony-Stimulating Factor In Vitro and In Vivo  Francesca Mascia, Christophe.
Th17 Cytokines Stimulate CCL20 Expression in Keratinocytes In Vitro and In Vivo: Implications for Psoriasis Pathogenesis  Erin G. Harper, Changsheng Guo,

Th17 Cytokines Stimulate CCL20 Expression in Keratinocytes In Vitro and In Vivo: Implications for Psoriasis Pathogenesis  Erin G. Harper, Changsheng Guo,
Coordinated Induction of Inducible Nitric Oxide Synthase and GTP-Cyclohydrolase I Is Dependent on Inflammatory Cytokines and Interferon-γ in HaCaT Keratinocytes:

Coordinated Induction of Inducible Nitric Oxide Synthase and GTP-Cyclohydrolase I Is Dependent on Inflammatory Cytokines and Interferon-γ in HaCaT Keratinocytes:

Similar presentations

Ads by Google