The IL-17F signaling pathway is involved in the induction of IFN-γ–inducible protein 10 in bronchial epithelial cells Mio Kawaguchi, MD, Fumio Kokubu,

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Presentation transcript:

The IL-17F signaling pathway is involved in the induction of IFN-γ–inducible protein 10 in bronchial epithelial cells Mio Kawaguchi, MD, Fumio Kokubu, MD, Shau-Ku Huang, PhD, Tetsuya Homma, MD, Miho Odaka, MD, Shin Watanabe, MD, Shintaro Suzuki, MD, Koushi Ieki, MD, Satoshi Matsukura, MD, Masatsugu Kurokawa, MD, Mitsuru Adachi, MD Journal of Allergy and Clinical Immunology Volume 119, Issue 6, Pages 1408-1414 (June 2007) DOI: 10.1016/j.jaci.2007.02.036 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 A, IP-10 gene expression by IL-17F in NHBEs. ∗P < .05 versus control; ∗∗P < .05 versus 10 ng/mL IL-17F–stimulated cells (n = 3). B, Time course of IP-10 gene expression. ∗P < .05 versus the level at the 0.5-hour time point (n = 3). C and D, IP-10 protein levels in lysates (Fig 1, C) and supernatants (Fig 1, D). ∗P < .05 versus medium control; ∗∗P < .05 versus 10 ng/mL IL-17F–stimulated cells (n = 6). The values are expressed as means ± SD. Journal of Allergy and Clinical Immunology 2007 119, 1408-1414DOI: (10.1016/j.jaci.2007.02.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Effect of other cytokines on IL-17F–induced IP-10 expression. NHBEs were stimulated as indicated, and IP-10 protein levels in supernatants were measured. The values are expressed as means ± SD (n = 3). ∗P < .05 versus control; ∗∗P < .05 versus IL-17F–stimulated cells. Journal of Allergy and Clinical Immunology 2007 119, 1408-1414DOI: (10.1016/j.jaci.2007.02.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Effect of PD98059 (PD), U0126, Raf1 kinase inhibitor I (Raf I), SB202190, and SP600125 on IP-10 gene (A) and protein (B) expression in NHBEs. The values are expressed as means ± SD (n = 4). ∗P < .05 versus IL-17F–stimulated cells in the absence of the inhibitor; ∗∗P < .05 versus the presence of an individual inhibitor. Journal of Allergy and Clinical Immunology 2007 119, 1408-1414DOI: (10.1016/j.jaci.2007.02.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Effect of overexpression of Raf1 dominant-negative mutants on IP-10 gene (A) and protein (B) expression in BEAS-2B cells. The values are expressed as means ± SD (n = 3). ∗P < .05 versus IL-17F–stimulated cells without vector. Journal of Allergy and Clinical Immunology 2007 119, 1408-1414DOI: (10.1016/j.jaci.2007.02.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 A, Kinetic activation of p90RSK by IL-17F in NHBEs. Western blotting was performed with antibodies against p90RSK and p-p90RSK. B, Effect of PD98059 on IL-17F–induced phosphorylation of p90RSK. Western blotting analysis was performed with antibodies against p90RSK and p-p90RSK. The results shown are representative of 3 separate experiments. Journal of Allergy and Clinical Immunology 2007 119, 1408-1414DOI: (10.1016/j.jaci.2007.02.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Regulation of IL-17F–induced IP-10 by p90RSK. BEAS-2B cells were transfected with or without siRNA for p90RSK, and its blocking was validated by means of Western blotting. The results shown are representative of 3 separate experiments (A). The levels of IP-10 gene expression (B) and protein production (C) were measured. The values are expressed as means ± SD (n = 3). ∗P < .05 versus nontransfected cells. Journal of Allergy and Clinical Immunology 2007 119, 1408-1414DOI: (10.1016/j.jaci.2007.02.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 7 A, Kinetic activation of CREB by IL-17F in NHBEs. Western blotting was performed with antibodies against CREB and p-CREB. B, Effect of siRNA for p90RSK on IL-17F–induced phosphorylation of CREB in BEAS-2B cells. Western blotting analysis was performed with antibodies against CREB and p-CREB. The results shown are representative of 3 separate experiments. Journal of Allergy and Clinical Immunology 2007 119, 1408-1414DOI: (10.1016/j.jaci.2007.02.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 8 Regulation of IL-17F–induced IP-10 by siRNA for CREB. The validation of its blocking by siRNA for CREB was performed by using Western blotting. The results shown are representative of 3 separate experiments (A). The levels of IP-10 gene expression (B) and protein production (C) were measured. The values are expressed as means ± SD (n = 3). ∗P < .05 versus nontransfected cells. Journal of Allergy and Clinical Immunology 2007 119, 1408-1414DOI: (10.1016/j.jaci.2007.02.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions