ABSTRACT
No disponible
Subject(s)
Humans , Male , Female , Liver Transplantation/classification , Liver Transplantation/instrumentation , Tissue Donors , Living Donors/classification , Digestive System Surgical Procedures/classification , Digestive System Surgical Procedures/instrumentation , Bile Ducts, Extrahepatic/surgery , Bile Ducts, Extrahepatic/transplantationABSTRACT
BACKGROUND/AIMS: Graft versus host disease (GVHD) following histoincompatible bone marrow transplantation may be modelled experimentally using irradiated metallothionein promoter-H-2Kb transgenic mice (MET-Kb mice), reconstituted with syngeneic non-transgenic spleen and lymph node (LN) cells. In this model, inflammation peaks at 3 weeks post-reconstitution, but resolves by 3 months, and is focussed on portal tracts and bile ducts (BD). The aim of this study was to determine if transgene-expressing hepatocytes play a role in the immune response, why portal tracts are selectively targeted, and which cell types are involved. METHODS: Intrahepatic BD (IHBD) with attached hepatocytes, or extrahepatic BD (EHBD) devoid of hepatocytes, were isolated from MET-Kb mice and implanted under the kidney capsule of transgenic (syngeneic) and congenic (allogeneic) mice. Three weeks post-implantation, BD were scored histologically for rejection or survival, and stained for various cell-surface molecules. RESULTS: Generally, IHBD survived better than EHBD, and T cells were the predominant infiltrating cell type in both implants. Both types of implants undergoing rejection expressed intercellular adhesion molecule-1 (ICAM-1) and leukocyte function antigen-1 (LFA-1) at high density; BD and the underlying kidney parenchyma also expressed class I and II major histocompatibility complex (MHC). CONCLUSIONS: The rejection of both groups of implants by congenic recipients suggests that BD from MET-Kb mice express the transgene, but the reason for the selective targeting of portal tracts rather than transgene-expressing hepatocytes remains unclear. One possible explanation is that dendritic cells/antigen-presenting cells (DC/APC) in portal tracts, which express high levels of MHC and co-stimulatory molecules, are the primary targets, and that BD are infiltrated and destroyed as 'bystanders'.
Subject(s)
Bile Ducts, Extrahepatic/transplantation , Bile Ducts, Intrahepatic/transplantation , Graft vs Host Disease/etiology , Kidney/surgery , Animals , Bile Ducts, Extrahepatic/metabolism , Bile Ducts, Extrahepatic/pathology , Bile Ducts, Intrahepatic/metabolism , Bile Ducts, Intrahepatic/pathology , CD4-Positive T-Lymphocytes/pathology , CD8-Positive T-Lymphocytes/pathology , Graft Rejection/etiology , Graft Rejection/metabolism , Graft Rejection/pathology , Graft vs Host Disease/metabolism , Graft vs Host Disease/pathology , H-2 Antigens/metabolism , Histocompatibility Antigens Class I/metabolism , Histocompatibility Antigens Class II/metabolism , Immunoenzyme Techniques , Intercellular Adhesion Molecule-1/metabolism , Lymphocyte Function-Associated Antigen-1/metabolism , Mice , Mice, TransgenicABSTRACT
The present paper describes the establishment of a method for simultaneous culturing of biliary epithelial cells (BECs) from the gall bladder (GB), extrahepatic bile duct (EBD) and intrahepatic bile duct (IBD) of the hamster. GB, EBD and IBD were cut from the biliary tree after collagenase perfusion of the liver. These biliary segments were minced into fragments. The fragments were embedded in collagen gel and cultured in Dulbecco's modified Eagle medium/HamF12 medium containing 10% fetal bovine serum. The various cells subsequently spread from the fragments and formed cellular sheets. After the fragments and flattened cells were removed with the aid of a Pasteur pipette under phase-contrast microscopy, the sheets remaining were found to be composed of cuboidal cells. These cuboidal cells were shown to express gamma glutamyl transpeptidase and cytokeratin 7, which are known to be specific markers of BECs. Ultrastructurally, a large number of microvilli were observed on the luminal surface and junctional complex and interdigitation was identifiable on the lateral surfaces. BEC cultures were subcultured by digestion with collagenase and dispase and then dissociated by subsequent digestion in trypsin and ethylenediaminetetraacetic acid and then maintained on collagen gel for up to 8 weeks. After several passages, the BECs in culture eventually increased in size and showed vacuoles in the cytoplasm. They demonstrated irreversible growth arrest at 9 weeks. The BECs tended to form cystic structures when the BECs with collagen gel were transplanted into the interscapular fat pads of syngeneic hamsters. We established a method for culturing and transplanting biliary cells from syrian golden hamsters. This method may help to clarify the mechanism of hepatobiliary diseases.
