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production of this cytokine, however, is associated with skin cancer development. Plant compounds have been shown to prevent chronic inflammation and cancer development in skin by suppressing the pathophysiological production of IL-1a in vivo [4,5]. An in vitro UVB cell model has also been developed to monitor the anti-inflammatory efficacy of plants and their compounds against IL-a for skin che- moprevention [6]. One of the plants that has been identified for potential use against skin inflammation during chemoprevention is Salsola tuberculata, due the potent anti- inflammatory activity of a synthetic analogue, Compound A [7,8]. However, the anti- inflammatory properties of S. tuberculata in skin still need to be elucidated. Aim: To investigate the effect of S. tuberculata extracts on icIL-1a production in the in vitro UVB skin cell model of inflammation and chemoprevention and compare activity to Compound A. Methods: Dichloromethane (DCM) and methanol extracts were prepared from S. tuberculata. The extracts and Compound A were screened for activity (P450 enzyme spectral assay) and cytotoxicity (cell viability assay). Sub-cytotoxic concentrations (IC 50 ) of the extracts and Compound A were used to determine the effect on icIL-1a production, cell viability, proliferation and apoptosis. Results: The DCM extract had no significant effect in cells. The methanol extract sig- nificantly (P < 0.05) enhanced UVB induced-icIL-1a production and promoted cyto- toxicity. However, the methanol extract had no significant effect on icIL-1a production in non-exposed cells. In contrast, Compound A inhibited UVB induced icIL-1a but also enhanced cytotoxicity. Conclusion: Contrary to the anti-inflammatory properties of its synthetic analogue (Compound A), S. tuberculata possesses pro-inflammatory properties that may not be effective against the early stages of inflammation in skin chemoprevention. References [1] Dinarello CA. Biologic basis for interleukin-1 in disease. Blood 1996;87(6):2095–147. [2] Lee WY, Lockniskar MF, Fischer SM. Interleukin-1 alpha mediates phorbol ester- induced inflammation and epidermal hyperplasia. Faseb J 1994;8(13):1081–7. [3] Li X, Eckard J, Shah R, Malluck C, Frenkel K. Interleukin-1alpha up-regulation in vivo by a potent carcinogen 7, 12-dimethylbenz (a) anthracene (DMBA) and control of DMBA-induced inflammatory responses. Can Res 2002;62(2):417–23. [4] Katiyar SK, Rupp CO, Korman NJ, Agarwal R, Mukhtar H. Inhibition of 12-O- tetradecanoylphorbol-13-acetate and other skin tumor-promoter-caused induc- tion of epidermal interleukin-1 alpha mRNA and protein expression in SENCAR mice by green tea polyphenols. J Invest Dermat 1995;105(3):394–8. [5] Katiyar SK, Mukhtar H. Inhibition of phorbol ester tumor promoter 12-O- tetradecanoylphorbol-13-acetate-caused inflammatory responses in SENCAR mouse skin by black tea polyphenols. Carcinogenesis 1997;18(10):1911–6. [6] Magcwebeba T, Riedel S, Swanevelder S, Bouic P, Swart P, Gelderblom W. Interleukin-1a induction in human keratinocytes (HaCaT): an in vitro model for chemoprevention in skin. J Skin Can 2012;2012:393681. doi:http://dx.doi.org/ 10.1155/2012/393681. [7] Kowalczyk P, Kowalczyk MC, Junco JJ, Tolstykh O, Kinjo T, Truong H, et al.. The possible separation of 12-O-tetradecanoylphorbol-13-acetate-induced skin inflammation and hyperplasia by compound A. Mol Carcin 2013;52(6):488–96. [8] Swart P, Swart AC, Louw A, van der Merwe KJ. Biological activities of the shrub Salsola tuberculatiformis Botsch: contraceptive or stress alleviator? Bioessays 2003;25(6):612–9. http://dx.doi.org/10.1016/j.cyto.2014.07.128 122 Intestinal epithelium is an autonomous lambda interferon-dependent cell com- partment that is calibrated by intestinal microflora Tanel Mahlakoiv 1 , Pedro Hernandez 2 , Andreas Diefenbach 3 , Peter Staeheli 1 , 1 Institute for Virology, University Medical Center Freiburg, Freiburg, Germany, 2 Institute for Medical Microbiology and Hygiene, University Medical Center Freiburg, Freiburg, Germany, 3 Institute of Medical Microbiology and Hygiene, University of Mainz Medical Centre, Mainz, Germany Type I interferon (IFN-a/b) represents the key element of the antiviral defense mechanisms against most viruses, however, rotaviruses that infect the gut epithe- lium, display little sensitivity to type I IFN. Here, we report that the intestinal epithe- lium is a unique cell compartment in the organism that does not depend on type I IFN in antiviral defenses. Type I IFN was unable to induce antiviral gene expression in intestinal epithelial cells (IEC) that correlated well with low epithelial expression of both chains of the IFN-a/b receptor complex. In stark contrast, IECs strongly responded to IFN-k on baseline, upon IFN treatment and virus challenge. Commensal microflora was found to establish baseline IFN-k signaling in IECs and enhance resis- tance to enteric virus infection. In adult mice lacking functional receptors for IFN-k, human reovirus selectively replicated in IECs. By contrast, these cells remained virus-free in IFN-a/b-deficient mice that developed deadly systemic disease after oral reovirus infection. In suckling mice with IFN-k receptor deficiency, reovirus not only replicated in the gut epithelium but also infected epithelial cells lining the bile ducts, thereby inducing a fatal liver disease. Interestingly, in response to synthetic double- stranded RNA or enteric virus infection, gut epithelial cells readily produced IFN-k but not IFN-a or IFN-b. Taken together, it appears that the intestinal epithelium has evolved mechanisms to specifically produce and respond to IFN-k, whereas systemic antiviral defenses depend on IFN-a/b produced predominantly by lymphoid cells. Our data suggest that such strict separation of the two IFN systems has evolved to avoid unnecessarily frequent triggering of the IFN-a/b system by commensals, which would activate systemic IFN pathways and thereby induce exacerbated inflammation. http://dx.doi.org/10.1016/j.cyto.2014.07.129 123 Functional analysis of CISH (Cytokine inducible SH2-containing protein) Saeed Mahmoudi , Daniel McCulloch, Alister Ward, School of Medicine, Strategic Research Centre in Molecular and Medical Research, Deakin University, Geelong, Victoria, Australia Introduction: The evolution of multicellular organisms has required the develop- ment of systems that allow cells to respond to external signals. Cytokine signalling represents one of these, and is involved in key biological processes, including blood and immune development as well as function. The ability to subsequently extinguish these signals is important to avoid pathogenic outcomes. The Suppressor of cytokine signalling (Socs) family of proteins represents an important mechanism to achieve this. The Cytokine Inducible SH2 domain-containing (CISH) protein was the first iden- tified member of the Socs family. In vitro studies have suggested the involvement of CISH as a negative regulator of signalling by EPO, IL-2, GH and GCSF. Recent genetic studies have implicated CISH in the susceptibility of humans to infectious disease and mice to allergic airway inflammation, as well as for a zebrafish homologue (Cish.a) in regulating embryonic haematopoiesis. Aim: This study has employed CISH targeted mice as a relevant model to further investigate the role of CISH in regulating development and disease. Methods: Mice were generated from ES cells in which the Cish locus was targeted with LacZ. These have been analysed by b-galactosidase staining and histology. Results: b-gal staining of Cish Lacz /+ mice embryos revealed expression from 11.5 post-conception, continuing in adult mice, across a wide range of different organs. Among haematopoietic organs CISH expression was predominantly observed in the thymus. Among visceral organs, expression was observed in kidney, stomach and liver. CISH expression was also observed in lung, heart, and brain. Sex-specific changes in expression were noted. Conclusion: CISH expression in a wide range of organs is consistent with the broad expression of other cytokine receptor signalling components, including Jak2 and Stat5a/Stat5b. Positive b-gal staining in the liver, kidneys, thymus, and brain support a regulatory role for CISH in these organs to control relevant cytokine signalling, such as by growth hormone, erythropoietin, IL-4 and IL-2, through the Jak/Stat signalling pathway. http://dx.doi.org/10.1016/j.cyto.2014.07.130 124 Immunotolerance-inducing IDO-1 and Th1-cytokines are upregulated in subcu- taneous panniculitis-like T-cell lymphoma; signs of an immunosuppressive microenvironment Pilvi Maliniemi 1 , Sonja Hahtola 1 , Kristian Ovaska 2 , Leila Jeskanen 1 , Liisa Väkevä 1 , Kirsi Niiranen 1 , Rudolf Stadler 3 , David Michonneau 4 , Sylvie Fraitag 5 , Sampsa Hautaniemi 2 , Annamari Ranki 1 , 1 University of Helsinki, Helsinki, Finland, 2 Computational Systems Biology Laboratory, Institute of Biomedicine and Genome-Scale Biology Program, University of Helsinki, Helsinki, Finland, 3 Johannes-Wesling-Klinikum Minden, Akademisches Lehrkrankenhaus der Medizinischen Hochschule Hannover, Minden, Germany, 4 Département d’immunologie, Equipe Dynamique des réponses immunes, 25 Rue du Docteur Roux, Institut Pasteur, Paris, France, 5 hôpital Necker- Enfants-Malades, AP-HP, 149 Rue de Sèvres, d’anatomie et de cytologie pathologiques, Paris, France Aims: Subcutaneous panniculitis-like T cell lymphomas represent a rare and diffi- cult to diagnose entity of cutaneous T cell lymphomas. SPTL affects predominantly young adults and presents with multifocal subcutaneous nodules and frequently associated autoimmune features. The pathogenesis of SPTL is not completely under- stood. The aim of this study is to unravel molecular pathways critical to the pathogen- esis for the first time. Abstract / Cytokine 70 (2014) 28–79 57
Transcript of 123
production of this cytokine, however, is associated with skin cancer development.Plant compounds have been shown to prevent chronic inflammation and cancerdevelopment in skin by suppressing the pathophysiological production of IL-1ain vivo [4,5]. An in vitro UVB cell model has also been developed to monitor theanti-inflammatory efficacy of plants and their compounds against IL-a for skin che-moprevention [6]. One of the plants that has been identified for potential use againstskin inflammation during chemoprevention is Salsola tuberculata, due the potent anti-inflammatory activity of a synthetic analogue, Compound A [7,8]. However, the anti-inflammatory properties of S. tuberculata in skin still need to be elucidated.
Aim: To investigate the effect of S. tuberculata extracts on icIL-1a production in thein vitro UVB skin cell model of inflammation and chemoprevention and compareactivity to Compound A.
Methods: Dichloromethane (DCM) and methanol extracts were prepared from S.tuberculata. The extracts and Compound A were screened for activity (P450 enzymespectral assay) and cytotoxicity (cell viability assay). Sub-cytotoxic concentrations(IC50) of the extracts and Compound A were used to determine the effect on icIL-1aproduction, cell viability, proliferation and apoptosis.
Results: The DCM extract had no significant effect in cells. The methanol extract sig-nificantly (P < 0.05) enhanced UVB induced-icIL-1a production and promoted cyto-toxicity. However, the methanol extract had no significant effect on icIL-1aproduction in non-exposed cells. In contrast, Compound A inhibited UVB inducedicIL-1a but also enhanced cytotoxicity.
Conclusion: Contrary to the anti-inflammatory properties of its synthetic analogue(Compound A), S. tuberculata possesses pro-inflammatory properties that may not beeffective against the early stages of inflammation in skin chemoprevention.
References
[1] Dinarello CA. Biologic basis for interleukin-1 in disease. Blood1996;87(6):2095–147.
[2] Lee WY, Lockniskar MF, Fischer SM. Interleukin-1 alpha mediates phorbol ester-induced inflammation and epidermal hyperplasia. Faseb J 1994;8(13):1081–7.
[3] Li X, Eckard J, Shah R, Malluck C, Frenkel K. Interleukin-1alpha up-regulationin vivo by a potent carcinogen 7, 12-dimethylbenz (a) anthracene (DMBA) andcontrol of DMBA-induced inflammatory responses. Can Res 2002;62(2):417–23.
[4] Katiyar SK, Rupp CO, Korman NJ, Agarwal R, Mukhtar H. Inhibition of 12-O-tetradecanoylphorbol-13-acetate and other skin tumor-promoter-caused induc-tion of epidermal interleukin-1 alpha mRNA and protein expression in SENCARmice by green tea polyphenols. J Invest Dermat 1995;105(3):394–8.
[5] Katiyar SK, Mukhtar H. Inhibition of phorbol ester tumor promoter 12-O-tetradecanoylphorbol-13-acetate-caused inflammatory responses in SENCARmouse skin by black tea polyphenols. Carcinogenesis 1997;18(10):1911–6.
[6] Magcwebeba T, Riedel S, Swanevelder S, Bouic P, Swart P, Gelderblom W.Interleukin-1a induction in human keratinocytes (HaCaT): an in vitro model forchemoprevention in skin. J Skin Can 2012;2012:393681. doi:http://dx.doi.org/10.1155/2012/393681.
[7] Kowalczyk P, Kowalczyk MC, Junco JJ, Tolstykh O, Kinjo T, Truong H, et al.. Thepossible separation of 12-O-tetradecanoylphorbol-13-acetate-induced skininflammation and hyperplasia by compound A. Mol Carcin 2013;52(6):488–96.
[8] Swart P, Swart AC, Louw A, van der Merwe KJ. Biological activities of the shrubSalsola tuberculatiformis Botsch: contraceptive or stress alleviator? Bioessays2003;25(6):612–9.
http://dx.doi.org/10.1016/j.cyto.2014.07.128
122Intestinal epithelium is an autonomous lambda interferon-dependent cell com-partment that is calibrated by intestinal microflora
Tanel Mahlakoiv 1, Pedro Hernandez 2, Andreas Diefenbach 3, Peter Staeheli 1,1 Institute for Virology, University Medical Center Freiburg, Freiburg, Germany,2 Institute for Medical Microbiology and Hygiene, University Medical Center Freiburg,Freiburg, Germany, 3 Institute of Medical Microbiology and Hygiene, University of MainzMedical Centre, Mainz, Germany
Type I interferon (IFN-a/b) represents the key element of the antiviral defensemechanisms against most viruses, however, rotaviruses that infect the gut epithe-lium, display little sensitivity to type I IFN. Here, we report that the intestinal epithe-lium is a unique cell compartment in the organism that does not depend on type I IFNin antiviral defenses. Type I IFN was unable to induce antiviral gene expression inintestinal epithelial cells (IEC) that correlated well with low epithelial expression ofboth chains of the IFN-a/b receptor complex. In stark contrast, IECs stronglyresponded to IFN-k on baseline, upon IFN treatment and virus challenge. Commensalmicroflora was found to establish baseline IFN-k signaling in IECs and enhance resis-tance to enteric virus infection. In adult mice lacking functional receptors for IFN-k,human reovirus selectively replicated in IECs. By contrast, these cells remained
virus-free in IFN-a/b-deficient mice that developed deadly systemic disease after oralreovirus infection. In suckling mice with IFN-k receptor deficiency, reovirus not onlyreplicated in the gut epithelium but also infected epithelial cells lining the bile ducts,thereby inducing a fatal liver disease. Interestingly, in response to synthetic double-stranded RNA or enteric virus infection, gut epithelial cells readily produced IFN-k butnot IFN-a or IFN-b. Taken together, it appears that the intestinal epithelium hasevolved mechanisms to specifically produce and respond to IFN-k, whereas systemicantiviral defenses depend on IFN-a/b produced predominantly by lymphoid cells. Ourdata suggest that such strict separation of the two IFN systems has evolved to avoidunnecessarily frequent triggering of the IFN-a/b system by commensals, which wouldactivate systemic IFN pathways and thereby induce exacerbated inflammation.
http://dx.doi.org/10.1016/j.cyto.2014.07.129
123Functional analysis of CISH (Cytokine inducible SH2-containing protein)
Saeed Mahmoudi, Daniel McCulloch, Alister Ward, School of Medicine, StrategicResearch Centre in Molecular and Medical Research, Deakin University, Geelong, Victoria,Australia
Introduction: The evolution of multicellular organisms has required the develop-ment of systems that allow cells to respond to external signals. Cytokine signallingrepresents one of these, and is involved in key biological processes, including bloodand immune development as well as function. The ability to subsequently extinguishthese signals is important to avoid pathogenic outcomes. The Suppressor of cytokinesignalling (Socs) family of proteins represents an important mechanism to achievethis. The Cytokine Inducible SH2 domain-containing (CISH) protein was the first iden-tified member of the Socs family. In vitro studies have suggested the involvement ofCISH as a negative regulator of signalling by EPO, IL-2, GH and GCSF. Recent geneticstudies have implicated CISH in the susceptibility of humans to infectious diseaseand mice to allergic airway inflammation, as well as for a zebrafish homologue(Cish.a) in regulating embryonic haematopoiesis.
Aim: This study has employed CISH targeted mice as a relevant model to furtherinvestigate the role of CISH in regulating development and disease.
Methods: Mice were generated from ES cells in which the Cish locus was targetedwith LacZ. These have been analysed by b-galactosidase staining and histology.
Results: b-gal staining of Cish Lacz �/+ mice embryos revealed expression from 11.5post-conception, continuing in adult mice, across a wide range of different organs.Among haematopoietic organs CISH expression was predominantly observed in thethymus. Among visceral organs, expression was observed in kidney, stomach andliver. CISH expression was also observed in lung, heart, and brain. Sex-specificchanges in expression were noted.
Conclusion: CISH expression in a wide range of organs is consistent with the broadexpression of other cytokine receptor signalling components, including Jak2 andStat5a/Stat5b. Positive b-gal staining in the liver, kidneys, thymus, and brain supporta regulatory role for CISH in these organs to control relevant cytokine signalling, suchas by growth hormone, erythropoietin, IL-4 and IL-2, through the Jak/Stat signallingpathway.
http://dx.doi.org/10.1016/j.cyto.2014.07.130
124Immunotolerance-inducing IDO-1 and Th1-cytokines are upregulated in subcu-taneous panniculitis-like T-cell lymphoma; signs of an immunosuppressivemicroenvironment
Pilvi Maliniemi 1, Sonja Hahtola 1, Kristian Ovaska 2, Leila Jeskanen 1, Liisa Väkevä 1,Kirsi Niiranen 1, Rudolf Stadler 3, David Michonneau 4, Sylvie Fraitag 5, SampsaHautaniemi 2, Annamari Ranki 1, 1 University of Helsinki, Helsinki, Finland,2 Computational Systems Biology Laboratory, Institute of Biomedicine and Genome-ScaleBiology Program, University of Helsinki, Helsinki, Finland, 3 Johannes-Wesling-KlinikumMinden, Akademisches Lehrkrankenhaus der Medizinischen Hochschule Hannover,Minden, Germany, 4 Département d’immunologie, Equipe Dynamique des réponsesimmunes, 25 Rue du Docteur Roux, Institut Pasteur, Paris, France, 5 hôpital Necker-Enfants-Malades, AP-HP, 149 Rue de Sèvres, d’anatomie et de cytologie pathologiques,Paris, France
Aims: Subcutaneous panniculitis-like T cell lymphomas represent a rare and diffi-cult to diagnose entity of cutaneous T cell lymphomas. SPTL affects predominantlyyoung adults and presents with multifocal subcutaneous nodules and frequentlyassociated autoimmune features. The pathogenesis of SPTL is not completely under-stood. The aim of this study is to unravel molecular pathways critical to the pathogen-esis for the first time.
Abstract / Cytokine 70 (2014) 28–79 57
