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IL-22/STAT3-Induced Increases in SLURP1 Expression within Psoriatic Lesions Exerts Antimicrobial Effects against Staphylococcus aureus.

Moriwaki Y, Takada K, Nagasaki T, Kubo N, Ishii T, Kose K, Kageyama T, Tsuji S, Kawashima K, Misawa H - PLoS ONE (2015)

Bottom Line: In NHEKs stimulated with the inflammatory cytokines IL-17, IL-22 and TNF-α, which are reportedly expressed in psoriatic lesions, SLURP1 mRNA expression was significantly up-regulated by IL-22 but not the other two cytokines.The stimulatory effect of IL-22 was completely suppressed in NHEKs treated with a STAT3 inhibitor or transfected with siRNA targeting STAT3.SLURP1 significantly suppressed the growth of S. aureus.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Faculty of Pharmacy, Keio University, Minato-ku, Tokyo 105-8512, Japan.

ABSTRACT

Background: SLURP1 is the causal gene for Mal de Meleda (MDM), an autosomal recessive skin disorder characterized by diffuse palmoplantar keratoderma and transgressive keratosis. Moreover, although SLURP1 likely serves as an important proliferation/differentiation factor in keratinocytes, the possible relation between SLURP1 and other skin diseases, such as psoriasis and atopic dermatitis, has not been studied, and the pathophysiological control of SLURP1 expression in keratinocytes is largely unknown.

Objectives: Our aim was to examine the involvement of SLURP1 in the pathophysiology of psoriasis using an imiquimod (IMQ)-induced psoriasis model mice and normal human epidermal keratinocytes (NHEKs).

Results: SLURP1 expression was up-regulated in the skin of IMQ-induced psoriasis model mice. In NHEKs stimulated with the inflammatory cytokines IL-17, IL-22 and TNF-α, which are reportedly expressed in psoriatic lesions, SLURP1 mRNA expression was significantly up-regulated by IL-22 but not the other two cytokines. The stimulatory effect of IL-22 was completely suppressed in NHEKs treated with a STAT3 inhibitor or transfected with siRNA targeting STAT3. Because IL-22 induces production of antimicrobial proteins in epithelial cells, the antibacterial activity of SLURP1 was assessed against Staphylococcus aureus (S. aureus), which is known to be associated with disease severity in psoriasis. SLURP1 significantly suppressed the growth of S. aureus.

Conclusions: These results indicate SLURP1 participates in pathophysiology of psoriasis by regulating keratinocyte proliferation and differentiation, and by suppressing the growth of S. aureus.

No MeSH data available.


Related in: MedlinePlus

STAT3 knockdown abolishes IL-22-induced SLURP1 induction in NHEKs.(A) NHEKs were incubated for 24 h with IL-22 (50 ng/ml) in the presence or absence of the MAP kinase kinase inhibitor PD98059 (30 μM), the STAT3 inhibitor S3I-201 (50 μM) or the STAT5 inhibitor 573108 (100) μM), after which SLURP1 expression was analyzed using real-time PCR. As a control, 0.1% (v/v) DMSO was used. Bars depict the mean ± S.D. (n = 6). (B, C) NHEKs were transfected with 100 nM scrambled control, STAT3 or STAT5 siRNA. After incubating the transfectants for 48 h, IL-22 was added to a final concentration of 50 ng/ml, and the cells were harvested 24 h later. (B) Cell lysates (30 μg of total protein/lane) were subjected to SDS-PAGE, and immunoblots were probed with anti-STAT3, anti-STAT5 or anti-β-actin antibody. The experiments were repeated six times with similar results. (C) Quantitative real-time PCR analysis of SLURP1 mRNA. Bars depict the mean ± S.D. (n = 6). ***p < 0.001 (one-way ANOVA).
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pone.0140750.g004: STAT3 knockdown abolishes IL-22-induced SLURP1 induction in NHEKs.(A) NHEKs were incubated for 24 h with IL-22 (50 ng/ml) in the presence or absence of the MAP kinase kinase inhibitor PD98059 (30 μM), the STAT3 inhibitor S3I-201 (50 μM) or the STAT5 inhibitor 573108 (100) μM), after which SLURP1 expression was analyzed using real-time PCR. As a control, 0.1% (v/v) DMSO was used. Bars depict the mean ± S.D. (n = 6). (B, C) NHEKs were transfected with 100 nM scrambled control, STAT3 or STAT5 siRNA. After incubating the transfectants for 48 h, IL-22 was added to a final concentration of 50 ng/ml, and the cells were harvested 24 h later. (B) Cell lysates (30 μg of total protein/lane) were subjected to SDS-PAGE, and immunoblots were probed with anti-STAT3, anti-STAT5 or anti-β-actin antibody. The experiments were repeated six times with similar results. (C) Quantitative real-time PCR analysis of SLURP1 mRNA. Bars depict the mean ± S.D. (n = 6). ***p < 0.001 (one-way ANOVA).

Mentions: To investigate the signal transduction pathway leading from IL-22 stimulation to enhanced SLURP1 transcription, IL-22-stimulated NHEKs were treated with a panel of signal transduction inhibitors. The enhancement of SLURP1 mRNA expression was completely blocked by the STAT3 inhibitor S3I-201, but was unaffected by the STAT5 inhibitor 573108 or the MAP kinase kinase inhibitor PD980589 (Fig 4A). To further confirm this result, NHEKs were transfected with siRNA targeting STAT3 or STAT5. Expression of the respective proteins was greatly suppressed in cells expressing STAT3 or STAT5 siRNA (Fig 4B). Moreover, IL-22-induced expression of SLURP1 mRNA was also suppressed in the STAT3 knockdown cells (Fig 4C). On the other hand, SLURP1 induction was unaffected by STAT5 knockdown. These results suggest that SLURP1 expression is under the control of the IL-22-STAT3 axis.


IL-22/STAT3-Induced Increases in SLURP1 Expression within Psoriatic Lesions Exerts Antimicrobial Effects against Staphylococcus aureus.

Moriwaki Y, Takada K, Nagasaki T, Kubo N, Ishii T, Kose K, Kageyama T, Tsuji S, Kawashima K, Misawa H - PLoS ONE (2015)

STAT3 knockdown abolishes IL-22-induced SLURP1 induction in NHEKs.(A) NHEKs were incubated for 24 h with IL-22 (50 ng/ml) in the presence or absence of the MAP kinase kinase inhibitor PD98059 (30 μM), the STAT3 inhibitor S3I-201 (50 μM) or the STAT5 inhibitor 573108 (100) μM), after which SLURP1 expression was analyzed using real-time PCR. As a control, 0.1% (v/v) DMSO was used. Bars depict the mean ± S.D. (n = 6). (B, C) NHEKs were transfected with 100 nM scrambled control, STAT3 or STAT5 siRNA. After incubating the transfectants for 48 h, IL-22 was added to a final concentration of 50 ng/ml, and the cells were harvested 24 h later. (B) Cell lysates (30 μg of total protein/lane) were subjected to SDS-PAGE, and immunoblots were probed with anti-STAT3, anti-STAT5 or anti-β-actin antibody. The experiments were repeated six times with similar results. (C) Quantitative real-time PCR analysis of SLURP1 mRNA. Bars depict the mean ± S.D. (n = 6). ***p < 0.001 (one-way ANOVA).
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pone.0140750.g004: STAT3 knockdown abolishes IL-22-induced SLURP1 induction in NHEKs.(A) NHEKs were incubated for 24 h with IL-22 (50 ng/ml) in the presence or absence of the MAP kinase kinase inhibitor PD98059 (30 μM), the STAT3 inhibitor S3I-201 (50 μM) or the STAT5 inhibitor 573108 (100) μM), after which SLURP1 expression was analyzed using real-time PCR. As a control, 0.1% (v/v) DMSO was used. Bars depict the mean ± S.D. (n = 6). (B, C) NHEKs were transfected with 100 nM scrambled control, STAT3 or STAT5 siRNA. After incubating the transfectants for 48 h, IL-22 was added to a final concentration of 50 ng/ml, and the cells were harvested 24 h later. (B) Cell lysates (30 μg of total protein/lane) were subjected to SDS-PAGE, and immunoblots were probed with anti-STAT3, anti-STAT5 or anti-β-actin antibody. The experiments were repeated six times with similar results. (C) Quantitative real-time PCR analysis of SLURP1 mRNA. Bars depict the mean ± S.D. (n = 6). ***p < 0.001 (one-way ANOVA).
Mentions: To investigate the signal transduction pathway leading from IL-22 stimulation to enhanced SLURP1 transcription, IL-22-stimulated NHEKs were treated with a panel of signal transduction inhibitors. The enhancement of SLURP1 mRNA expression was completely blocked by the STAT3 inhibitor S3I-201, but was unaffected by the STAT5 inhibitor 573108 or the MAP kinase kinase inhibitor PD980589 (Fig 4A). To further confirm this result, NHEKs were transfected with siRNA targeting STAT3 or STAT5. Expression of the respective proteins was greatly suppressed in cells expressing STAT3 or STAT5 siRNA (Fig 4B). Moreover, IL-22-induced expression of SLURP1 mRNA was also suppressed in the STAT3 knockdown cells (Fig 4C). On the other hand, SLURP1 induction was unaffected by STAT5 knockdown. These results suggest that SLURP1 expression is under the control of the IL-22-STAT3 axis.

Bottom Line: In NHEKs stimulated with the inflammatory cytokines IL-17, IL-22 and TNF-α, which are reportedly expressed in psoriatic lesions, SLURP1 mRNA expression was significantly up-regulated by IL-22 but not the other two cytokines.The stimulatory effect of IL-22 was completely suppressed in NHEKs treated with a STAT3 inhibitor or transfected with siRNA targeting STAT3.SLURP1 significantly suppressed the growth of S. aureus.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Faculty of Pharmacy, Keio University, Minato-ku, Tokyo 105-8512, Japan.

ABSTRACT

Background: SLURP1 is the causal gene for Mal de Meleda (MDM), an autosomal recessive skin disorder characterized by diffuse palmoplantar keratoderma and transgressive keratosis. Moreover, although SLURP1 likely serves as an important proliferation/differentiation factor in keratinocytes, the possible relation between SLURP1 and other skin diseases, such as psoriasis and atopic dermatitis, has not been studied, and the pathophysiological control of SLURP1 expression in keratinocytes is largely unknown.

Objectives: Our aim was to examine the involvement of SLURP1 in the pathophysiology of psoriasis using an imiquimod (IMQ)-induced psoriasis model mice and normal human epidermal keratinocytes (NHEKs).

Results: SLURP1 expression was up-regulated in the skin of IMQ-induced psoriasis model mice. In NHEKs stimulated with the inflammatory cytokines IL-17, IL-22 and TNF-α, which are reportedly expressed in psoriatic lesions, SLURP1 mRNA expression was significantly up-regulated by IL-22 but not the other two cytokines. The stimulatory effect of IL-22 was completely suppressed in NHEKs treated with a STAT3 inhibitor or transfected with siRNA targeting STAT3. Because IL-22 induces production of antimicrobial proteins in epithelial cells, the antibacterial activity of SLURP1 was assessed against Staphylococcus aureus (S. aureus), which is known to be associated with disease severity in psoriasis. SLURP1 significantly suppressed the growth of S. aureus.

Conclusions: These results indicate SLURP1 participates in pathophysiology of psoriasis by regulating keratinocyte proliferation and differentiation, and by suppressing the growth of S. aureus.

No MeSH data available.


Related in: MedlinePlus