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O028 The signaling properties of Interleukin-30 underline plasticity and cross-talk among the Interleukin-6 family of cytokines
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Abstract / Cytokine 59 (2012) 509–510
Methods. Mice carrying epidermal-specific deletion of Prdm1, the gene encoding Blimp-1, in adult stage in an inducible manner were generated for this study. Results. We first show that Blimp-1 expression is reduced in stimulated mouse keratinocytes. Epidermal-specific deletion of Prdm1 in adult mice caused spontaneously inflamed skin characterized by massive dermal infiltration of neutrophils/macrophages and development of chronic inflammation, leading to increased release of cytokines/chemokines, including G-CSF, and enhanced granulopoiesis in bone marrow. Deletion of Blimp-1 in the epidermis of adult mice also led to sensitized inflammatory reactions in a disease model of contact dermatitis. Systemic increases in G-CSF contributed to the inflammatory responses because neutralization of G-CSF or deletion of the G-CSF gene, Csf3, corrected neutrophilia and partially ameliorated the inflamed skin in Prdm1-deficient mice. Conclusion. We show a novel function for Blimp-1 acting in repressing cytokine/ chemokine genes in restraining steady-state epidermal immune reactions. Disclosure of interest: None declared. http://dx.doi.org/10.1016/j.cyto.2012.06.060
O028 The signaling properties of Interleukin-30 underline plasticity and cross-talk among the Interleukin-6 family of cytokines C. Garbers 1, S. Aparicio-Siegmund 1, B. Spudy 2, G.H. Waetzig 3, I. Lorenzen 2, J. Grötzinger 2, S. Rose-John 2, J. Scheller 1, 1 Institute of Biochemistry and Molecular Biology II, Heinrich-Heine-University of Duesseldorf, Duesseldorf, Germany, 2 Institute of Biochemistry, Christian-Albrechts-University of Kiel, Germany, 3 Conaris Research Institute AG, Kiel, Germany Introduction . The anti-inflammatory Interleukin (IL)-6/IL-12 cytokine family member IL-27 consists of the cytokine Interleukin-30 (IL-30, also referred to as IL-27p28)
and the non-signaling a-receptor subunit Epstein-Barr virus induced gene 3 (EBI3). Antigen-presenting cells are the major cellular source of both proteins, and IL-27 has been shown to engage signaling via a heterodimer of the two transmembrane receptors WSX-1 and gp130. Besides this well-established function, several studies suggest that IL-30 has signaling properties on its own. Recently, IL-30 was shown to form a novel cytokine complex with the non-signaling membrane-bound IL-6 receptor a (IL-6R). Methods. Since free, endogenous IL-30 is not efficiently secreted, we developed a strategy for bacterial expression, purification and renaturation of murine IL-30 (mIL-30) for in vitro and in vivo analysis. Furthermore, we designed a Hyper-cytokine, consisting of the extracellular/soluble part of the IL-6R fused via a peptide linker to IL30, and expressed the IL-30-sIL-6R fusion protein in mammalian cells. Results. We show that mIL-30 can induce STAT-dependent proliferation of Ba/F3gp130 cells expressing murine or human IL-6R, indicating that in contrast to murine IL-6, murine IL-30 possesses no species specificity. Furthermore, we identified the two transmembrane proteins that function as the signal transducing receptors of the IL-30/IL-6R complex. IL-30 was also able to induce IL-30-trans-signaling using the soluble IL-6R (sIL-6R), a characteristic property that was initially described for IL-6 signaling via the sIL-6R. We further show how STAT phosphorylation and proliferation of Ba/F3-gp130 cells induced by IL-30-sIL-6R can be efficiently inhibited. Finally, mIL-30 but not IL-6, was able to induce STAT-dependent proliferation of Ba/F3-gp130 cells without the need of the p28-a receptors IL-6R and EBI3, albeit at higher concentrations of mIL-30 in comparison to IL-30-sIL-6R. Conclusion. Our study enlarges the spectrum of IL-30-dependent receptor activation pathways and might have major implications for the biological role of Interleukin-30 in vivo. Disclosure of interest: None declared. http://dx.doi.org/10.1016/j.cyto.2012.06.061
Abstract / Cytokine 59 (2012) 509–510
Methods. Mice carrying epidermal-specific deletion of Prdm1, the gene encoding Blimp-1, in adult stage in an inducible manner were generated for this study. Results. We first show that Blimp-1 expression is reduced in stimulated mouse keratinocytes. Epidermal-specific deletion of Prdm1 in adult mice caused spontaneously inflamed skin characterized by massive dermal infiltration of neutrophils/macrophages and development of chronic inflammation, leading to increased release of cytokines/chemokines, including G-CSF, and enhanced granulopoiesis in bone marrow. Deletion of Blimp-1 in the epidermis of adult mice also led to sensitized inflammatory reactions in a disease model of contact dermatitis. Systemic increases in G-CSF contributed to the inflammatory responses because neutralization of G-CSF or deletion of the G-CSF gene, Csf3, corrected neutrophilia and partially ameliorated the inflamed skin in Prdm1-deficient mice. Conclusion. We show a novel function for Blimp-1 acting in repressing cytokine/ chemokine genes in restraining steady-state epidermal immune reactions. Disclosure of interest: None declared. http://dx.doi.org/10.1016/j.cyto.2012.06.060
O028 The signaling properties of Interleukin-30 underline plasticity and cross-talk among the Interleukin-6 family of cytokines C. Garbers 1, S. Aparicio-Siegmund 1, B. Spudy 2, G.H. Waetzig 3, I. Lorenzen 2, J. Grötzinger 2, S. Rose-John 2, J. Scheller 1, 1 Institute of Biochemistry and Molecular Biology II, Heinrich-Heine-University of Duesseldorf, Duesseldorf, Germany, 2 Institute of Biochemistry, Christian-Albrechts-University of Kiel, Germany, 3 Conaris Research Institute AG, Kiel, Germany Introduction . The anti-inflammatory Interleukin (IL)-6/IL-12 cytokine family member IL-27 consists of the cytokine Interleukin-30 (IL-30, also referred to as IL-27p28)
and the non-signaling a-receptor subunit Epstein-Barr virus induced gene 3 (EBI3). Antigen-presenting cells are the major cellular source of both proteins, and IL-27 has been shown to engage signaling via a heterodimer of the two transmembrane receptors WSX-1 and gp130. Besides this well-established function, several studies suggest that IL-30 has signaling properties on its own. Recently, IL-30 was shown to form a novel cytokine complex with the non-signaling membrane-bound IL-6 receptor a (IL-6R). Methods. Since free, endogenous IL-30 is not efficiently secreted, we developed a strategy for bacterial expression, purification and renaturation of murine IL-30 (mIL-30) for in vitro and in vivo analysis. Furthermore, we designed a Hyper-cytokine, consisting of the extracellular/soluble part of the IL-6R fused via a peptide linker to IL30, and expressed the IL-30-sIL-6R fusion protein in mammalian cells. Results. We show that mIL-30 can induce STAT-dependent proliferation of Ba/F3gp130 cells expressing murine or human IL-6R, indicating that in contrast to murine IL-6, murine IL-30 possesses no species specificity. Furthermore, we identified the two transmembrane proteins that function as the signal transducing receptors of the IL-30/IL-6R complex. IL-30 was also able to induce IL-30-trans-signaling using the soluble IL-6R (sIL-6R), a characteristic property that was initially described for IL-6 signaling via the sIL-6R. We further show how STAT phosphorylation and proliferation of Ba/F3-gp130 cells induced by IL-30-sIL-6R can be efficiently inhibited. Finally, mIL-30 but not IL-6, was able to induce STAT-dependent proliferation of Ba/F3-gp130 cells without the need of the p28-a receptors IL-6R and EBI3, albeit at higher concentrations of mIL-30 in comparison to IL-30-sIL-6R. Conclusion. Our study enlarges the spectrum of IL-30-dependent receptor activation pathways and might have major implications for the biological role of Interleukin-30 in vivo. Disclosure of interest: None declared. http://dx.doi.org/10.1016/j.cyto.2012.06.061