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Fas ligand induced T-cell lymphocyte apoptosis
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Abstracts / Digestive and Liver Disease 46 (2014) e18–e42
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HUMAN BILIARY TREE STEM/PROGENITOR CELLS (HBTSCS) FROM PERIBILIARY GLANDS (PBGS) OF ADULT LIVER DISPLAY IMMUNOMODULATORY PROPERTIES THROUGH FAS/FAS LIGAND INDUCED T-CELL LYMPHOCYTE APOPTOSIS
SEX HORMONES DIFFERENTLY REGULATE HEPATIC HEPCIDIN EXPRESSION AND SYSTEMIC IRON HOMEOSTASIS IN VIVO
G. Carnevale 1 , M. Riccio 1 , V. Cardinale 2 , L. Gibelini 1 , S. De Biasi 1 , A. Pisciotta 1 , G. Carpino 3 , R. Gentile 2 , P.B. Berloco 4 , R. Brunelli 5 , C. Bastianelli 5 , A. Cossarizza 1 , E. Gaudio 6 , D. Alvaro 2 , A. De Pol 1 1
Department of Surgery, Medicine, Dentistry and Morphological Sciences, University of Modena and Reggio Emilia, Modena, Italy 2 Department of Medico-Surgical Sciences and Biotechnologies, Polo Pontino, Sapienza University of Rome, Rome, Italy 3 Department of Movement, Human and Health Sciences, Division of Health Sciences, University of Rome “Foro Italico”, Rome, Italy 4 Department of General Surgery and Organ Transplantation, Sapienza University of Rome, Rome, Italy 5 Department of Obstetrics and Gynecology, Sapienza University of Rome, Rome, Italy 6 Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy Introduction: hBTSCs have the potential for regenerative medicine in liver and pancreas diseases. T-cell control was recently demonstrated for mesenchymal stem cells. Aim: The aims of this study w ere to evaluate Fas-L expression within the stem cell niches of adult biliary tree (PBGs), and to study the interaction between hBTSCs and human lymphocytes. Materials and methods: HLA antigens, Fas and Fas-L expression were evaluated by immunofluorescence and Western blotting (WB) in cells of human biliary tree in comparison with fibroblast cells, dental pulp stem cells and bone marrow mesenchymal stem cells. The influence of hBTSCs on lymphocytes’ activation and apoptosis w ere assessed by co-culturing experiments. Results: Adult hBTSCs expressed both class I and class II HLA antigens, whereas fetal hBTSCs only class I HLA antigens. 10 to 30% of the hBTSCs in PBGs were positive for Fas-L. Fas-L+ cells were mostly located at the bottom of PBGs and co-expressed EpCAM (Epithelial-Cell-Adhesion-Molecule) and proliferation marker (PCNA: Proliferating-Cell-Nuclear-Antigen). Mature cells at the bile duct surface epithelium (mature cholangiocytes) were almost all negative for Fas-L. In culture experiments confocal microscopy demonstrated that Fas-L expression was restricted to EpCAM+/LGR5+ (a marker associated with endodermal stem cells) hBTSCs. WB confirmed that hBTSCs constitutively expressed high level of Fas-L which increased after co-culture with T-cells. Fluorescence-activated cell sorting (FACS) analysis revealed that activated CD4+ and CD8+ T-cells co-cultured with hBTSCs underwent to a massive induction of apoptosis. Fas receptor appeared over-expressed in T-cells co-cultured with hBTSCs respect to resting T-cells. Conclusions: Our data demonstrated that hBTSCs can induce “premature” apoptosis in T-cells trough the activation of Fas/Fas-L pathway. http://dx.doi.org/10.1016/j.dld.2014.01.069
C. Garuti 1,2 , G. Montosi 1,2 , S. Barelli 1,2 , A. Pietrangelo 1,2 , E. Corradini 1,2 1
Internal Medicine 2 and Center for Hemochromatosis, University Hospital of Modena, Italy 2 Department of Medical and Surgical Science for Children and Adults, University of Modena and Reggio Emilia, Italy Introduction: Iron loading has been associated with the progression of several liver diseases. Moreover, a number of studies indicate that female and male exhibit differences in the progression of chronic liver diseases, including hereditary hemochromatosis. Better understanding of the mechanisms associated with genderrelated differences in iron homeostasis may help to develop new therapeutic strategies in hepatic diseases. Hepcidin, a small hormone synthesized mainly by the liver, is the central regulator of iron homeostasis. The BMP(6)-SMAD signaling cascade is the main pathway involved in hepcidin expression regulation. In humans and animal models, gender influences hepcidin and body iron levels. So far data on the role of sex hormones in the control of hepcidin transcription and iron homeostasis have been controversial. Aim: To study the effects of androgens and estrogens on the regulation of hepcidin expression and systemic iron status in vivo. Materials and methods: Male and female mice underwent orchiectomy or ovariectomy. Furthermore, male and female mice were parenterally treated with androgens or estrogens. Results: Orchiectomized mice showed higher hepcidin transcription, a significant activation of the BMP6-SMAD pathway, and lower liver iron content in comparison to sham-operated mice. This was not the case for ovariectomized mice. Yet, androgens administration led to a 50% decrease of hepcidin mRNA and significant spleen iron depletion. In contrast, estrogens administration, while decreasing hepcidin transcription, was associated with spleen iron retention. Conclusions: Our data indicate that both male and female sex hormones control hepatic hepcidin expression; estrogens seem to act on systemic iron status not only via direct inhibition of hepcidin transcription but also through a non-hepcidin driven effect involving tissue iron re-distribution. http://dx.doi.org/10.1016/j.dld.2014.01.070 T-31 SIRT1 ACTIVITY IN HEPATIC STELLATE CELLS DURING LIVER INJURY IS A TARGET FOR THE MODULATION OF THE FIBROGENIC PROCESS M. Tarocchi 1 , G. Marroncini 2 , T. Mello 1 , S. Polvani 1 , S. Tempesti 1 , E. Ceni 1 , A. Galli 1,2 1 Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy 2 Pharmacogenomic Foundation FiorGen, Florence, Italy
Introduction: Hepatic stellate cells (HSC) represent 15% of the total resident cells in normal liver. Following liver injury, HSC transform into proliferating profibrogenic myofibroblasts. The activation of SIRT1, a NAD-dependent histone deacetylase, is correlated
Abstracts / Digestive and Liver Disease 46 (2014) e18–e42
T-29
T-30
HUMAN BILIARY TREE STEM/PROGENITOR CELLS (HBTSCS) FROM PERIBILIARY GLANDS (PBGS) OF ADULT LIVER DISPLAY IMMUNOMODULATORY PROPERTIES THROUGH FAS/FAS LIGAND INDUCED T-CELL LYMPHOCYTE APOPTOSIS
SEX HORMONES DIFFERENTLY REGULATE HEPATIC HEPCIDIN EXPRESSION AND SYSTEMIC IRON HOMEOSTASIS IN VIVO
G. Carnevale 1 , M. Riccio 1 , V. Cardinale 2 , L. Gibelini 1 , S. De Biasi 1 , A. Pisciotta 1 , G. Carpino 3 , R. Gentile 2 , P.B. Berloco 4 , R. Brunelli 5 , C. Bastianelli 5 , A. Cossarizza 1 , E. Gaudio 6 , D. Alvaro 2 , A. De Pol 1 1
Department of Surgery, Medicine, Dentistry and Morphological Sciences, University of Modena and Reggio Emilia, Modena, Italy 2 Department of Medico-Surgical Sciences and Biotechnologies, Polo Pontino, Sapienza University of Rome, Rome, Italy 3 Department of Movement, Human and Health Sciences, Division of Health Sciences, University of Rome “Foro Italico”, Rome, Italy 4 Department of General Surgery and Organ Transplantation, Sapienza University of Rome, Rome, Italy 5 Department of Obstetrics and Gynecology, Sapienza University of Rome, Rome, Italy 6 Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, Sapienza University of Rome, Rome, Italy Introduction: hBTSCs have the potential for regenerative medicine in liver and pancreas diseases. T-cell control was recently demonstrated for mesenchymal stem cells. Aim: The aims of this study w ere to evaluate Fas-L expression within the stem cell niches of adult biliary tree (PBGs), and to study the interaction between hBTSCs and human lymphocytes. Materials and methods: HLA antigens, Fas and Fas-L expression were evaluated by immunofluorescence and Western blotting (WB) in cells of human biliary tree in comparison with fibroblast cells, dental pulp stem cells and bone marrow mesenchymal stem cells. The influence of hBTSCs on lymphocytes’ activation and apoptosis w ere assessed by co-culturing experiments. Results: Adult hBTSCs expressed both class I and class II HLA antigens, whereas fetal hBTSCs only class I HLA antigens. 10 to 30% of the hBTSCs in PBGs were positive for Fas-L. Fas-L+ cells were mostly located at the bottom of PBGs and co-expressed EpCAM (Epithelial-Cell-Adhesion-Molecule) and proliferation marker (PCNA: Proliferating-Cell-Nuclear-Antigen). Mature cells at the bile duct surface epithelium (mature cholangiocytes) were almost all negative for Fas-L. In culture experiments confocal microscopy demonstrated that Fas-L expression was restricted to EpCAM+/LGR5+ (a marker associated with endodermal stem cells) hBTSCs. WB confirmed that hBTSCs constitutively expressed high level of Fas-L which increased after co-culture with T-cells. Fluorescence-activated cell sorting (FACS) analysis revealed that activated CD4+ and CD8+ T-cells co-cultured with hBTSCs underwent to a massive induction of apoptosis. Fas receptor appeared over-expressed in T-cells co-cultured with hBTSCs respect to resting T-cells. Conclusions: Our data demonstrated that hBTSCs can induce “premature” apoptosis in T-cells trough the activation of Fas/Fas-L pathway. http://dx.doi.org/10.1016/j.dld.2014.01.069
C. Garuti 1,2 , G. Montosi 1,2 , S. Barelli 1,2 , A. Pietrangelo 1,2 , E. Corradini 1,2 1
Internal Medicine 2 and Center for Hemochromatosis, University Hospital of Modena, Italy 2 Department of Medical and Surgical Science for Children and Adults, University of Modena and Reggio Emilia, Italy Introduction: Iron loading has been associated with the progression of several liver diseases. Moreover, a number of studies indicate that female and male exhibit differences in the progression of chronic liver diseases, including hereditary hemochromatosis. Better understanding of the mechanisms associated with genderrelated differences in iron homeostasis may help to develop new therapeutic strategies in hepatic diseases. Hepcidin, a small hormone synthesized mainly by the liver, is the central regulator of iron homeostasis. The BMP(6)-SMAD signaling cascade is the main pathway involved in hepcidin expression regulation. In humans and animal models, gender influences hepcidin and body iron levels. So far data on the role of sex hormones in the control of hepcidin transcription and iron homeostasis have been controversial. Aim: To study the effects of androgens and estrogens on the regulation of hepcidin expression and systemic iron status in vivo. Materials and methods: Male and female mice underwent orchiectomy or ovariectomy. Furthermore, male and female mice were parenterally treated with androgens or estrogens. Results: Orchiectomized mice showed higher hepcidin transcription, a significant activation of the BMP6-SMAD pathway, and lower liver iron content in comparison to sham-operated mice. This was not the case for ovariectomized mice. Yet, androgens administration led to a 50% decrease of hepcidin mRNA and significant spleen iron depletion. In contrast, estrogens administration, while decreasing hepcidin transcription, was associated with spleen iron retention. Conclusions: Our data indicate that both male and female sex hormones control hepatic hepcidin expression; estrogens seem to act on systemic iron status not only via direct inhibition of hepcidin transcription but also through a non-hepcidin driven effect involving tissue iron re-distribution. http://dx.doi.org/10.1016/j.dld.2014.01.070 T-31 SIRT1 ACTIVITY IN HEPATIC STELLATE CELLS DURING LIVER INJURY IS A TARGET FOR THE MODULATION OF THE FIBROGENIC PROCESS M. Tarocchi 1 , G. Marroncini 2 , T. Mello 1 , S. Polvani 1 , S. Tempesti 1 , E. Ceni 1 , A. Galli 1,2 1 Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy 2 Pharmacogenomic Foundation FiorGen, Florence, Italy
Introduction: Hepatic stellate cells (HSC) represent 15% of the total resident cells in normal liver. Following liver injury, HSC transform into proliferating profibrogenic myofibroblasts. The activation of SIRT1, a NAD-dependent histone deacetylase, is correlated