D Klein1, F Timoneri, H Ichii, C Ricordi, R L Pastori. 1. Diabetes Research Institute, Leonard M. Miller School of Medicine, University of Miami, 1450 NW 10th Avenue, Miami, FL 33136, USA.
Abstract
AIMS/HYPOTHESIS: CD40 expression on non-haematopoietic cells is linked to inflammation. We previously reported that CD40 is expressed on isolated human and non-human primate islets and its activation results in secretion of IL-8, macrophage inflammatory protein 1-beta (MIP-1beta) and monocyte chemoattractant protein-1 (MCP-1) through nuclear factor-kappaB and extracellularly regulated kinases 1/2 pathways. The objective of this study was to identify the pattern of gene expression, and to study viability and functionality affected by CD40-CD40 ligand (CD40L) interaction in human islets. Furthermore, we have studied the CD40-mediated cytokine/chemokine profile in pancreatic ductal cells, as they are always present in human islet transplant preparations and express CD40 constitutively. METHODS: CD40-CD40L gene expression modulation was studied by microarray on islet cells depleted of ductal cells. Selected genes were validated by quantitative RT-PCR. The cytokine profile in purified ductal cells was evaluated by Luminex technology, based on the use of fluorescent-coated beads, known as microspheres, and capable of multiplex detection of proteins from a single sample. Glucose-stimulated insulin secretion and islet viability were assessed by perifusion and 7-aminoactinomycin D membrane exclusion, respectively. RESULTS: Statistical analysis of microarrays identified 30 genes exhibiting at least a 2.5-fold increase across all replicate arrays. The majority of them were related to oxidative stress/inflammation. Prominently upregulated were chemokine C-X-C motif ligand 1 (CXCL1), CXCL2 and CXCL3 belonging to the CXC family of chemokines related to IL-8. CD40-mediated CXCL1 secretion was confirmed by ELISA. The viability or in vitro function was not affected by CD40 activation. In addition to previously reported IL-8, MIP-1beta and MCP-1, CD40 stimulation in ductal cells produced IL-1beta, IFN-gamma, TNF-alpha, granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. CONCLUSIONS/ INTERPRETATION: CD40 activation in islets and ductal cells produces cytokines/chemokines with a broad-spectrum range of biological functions.
AIMS/HYPOTHESIS: CD40 expression on non-haematopoietic cells is linked to inflammation. We previously reported that CD40 is expressed on isolated human and non-human primate islets and its activation results in secretion of IL-8, macrophage inflammatory protein 1-beta (MIP-1beta) and monocyte chemoattractant protein-1 (MCP-1) through nuclear factor-kappaB and extracellularly regulated kinases 1/2 pathways. The objective of this study was to identify the pattern of gene expression, and to study viability and functionality affected by CD40-CD40 ligand (CD40L) interaction in human islets. Furthermore, we have studied the CD40-mediated cytokine/chemokine profile in pancreatic ductal cells, as they are always present in human islet transplant preparations and express CD40 constitutively. METHODS:CD40-CD40L gene expression modulation was studied by microarray on islet cells depleted of ductal cells. Selected genes were validated by quantitative RT-PCR. The cytokine profile in purified ductal cells was evaluated by Luminex technology, based on the use of fluorescent-coated beads, known as microspheres, and capable of multiplex detection of proteins from a single sample. Glucose-stimulated insulin secretion and islet viability were assessed by perifusion and 7-aminoactinomycin D membrane exclusion, respectively. RESULTS: Statistical analysis of microarrays identified 30 genes exhibiting at least a 2.5-fold increase across all replicate arrays. The majority of them were related to oxidative stress/inflammation. Prominently upregulated were chemokine C-X-C motif ligand 1 (CXCL1), CXCL2 and CXCL3 belonging to the CXC family of chemokines related to IL-8. CD40-mediated CXCL1 secretion was confirmed by ELISA. The viability or in vitro function was not affected by CD40 activation. In addition to previously reported IL-8, MIP-1beta and MCP-1, CD40 stimulation in ductal cells produced IL-1beta, IFN-gamma, TNF-alpha, granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. CONCLUSIONS/ INTERPRETATION:CD40 activation in islets and ductal cells produces cytokines/chemokines with a broad-spectrum range of biological functions.
Authors: Edward B Alabraba; Vincent Lai; Louis Boon; Stephen J Wigmore; David H Adams; Simon C Afford Journal: Hepatology Date: 2008-02 Impact factor: 17.425
Authors: O Vosters; C Beuneu; N Nagy; B Movahedi; E Aksoy; I Salmon; D Pipeleers; M Goldman; V Verhasselt Journal: Diabetologia Date: 2004-04 Impact factor: 10.122
Authors: Thomas Enzler; Silke Gillessen; Michael Dougan; James P Allison; Donna Neuberg; Darryl A Oble; Martin Mihm; Glenn Dranoff Journal: Blood Date: 2007-05-04 Impact factor: 22.113
Authors: Arne Egesten; Mette Eliasson; Anders I Olin; Jonas S Erjefält; Anders Bjartell; Per Sangfelt; Marie Carlson Journal: Int J Colorectal Dis Date: 2007-08-17 Impact factor: 2.571
Authors: Pascale Ribaux; Jan A Ehses; Nathalie Lin-Marq; Fabio Carrozzino; Marianne Böni-Schnetzler; Eva Hammar; Jean-Claude Irminger; Marc Y Donath; Philippe A Halban Journal: Endocrinology Date: 2007-08-16 Impact factor: 4.736
Authors: Nikolaos T Georgopoulos; Alison Merrick; Nigel Scott; Peter J Selby; Alan Melcher; Ludwik K Trejdosiewicz Journal: Int J Cancer Date: 2007-09-15 Impact factor: 7.396
Authors: D Jacobs-Tulleneers-Thevissen; M Chintinne; Z Ling; P Gillard; L Schoonjans; G Delvaux; B L Strand; F Gorus; B Keymeulen; D Pipeleers Journal: Diabetologia Date: 2013-04-26 Impact factor: 10.122
Authors: Marta Bagnati; Babatunji W Ogunkolade; Catriona Marshall; Carmen Tucci; Katie Hanna; Tania A Jones; Marco Bugliani; Belinda Nedjai; Paul W Caton; Julius Kieswich; Muhammed M Yaqoob; Graham R Ball; Piero Marchetti; Graham A Hitman; Mark D Turner Journal: Cell Death Dis Date: 2016-08-11 Impact factor: 8.469