ABSTRACT
Epithelial cells are positioned in close proximity to endothelial cells. A non-contact coculture system was used to investigate whether colonic epithelial cells activated with various cytokines are able to provide signals that can modulate ICAM-1 and VCAM-1 expression on endothelial cells. Coculture of human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC-1) with TNF-alpha/IFN-gamma-stimulated human colon epithelial cell lines led to a significant up-regulation of endothelial ICAM-1 and VCAM-1 expression. Increased ICAM-1 and VCAM-1 expression by endothelial cells was accompanied by an increase in endothelial cell NF-kappaB p65 and NF-kappaB-DNA-binding activity. Inhibition of endothelial NF-kappaB activation using the proteosome inhibitors MG-132 and BAY 11-7082 resulted in a significant decrease of ICAM-1 expression, indicating an important role for NF-kappaB in this response. This cross-talk may represent a biological mechanism for the gut epithelium to control the colonic inflammatory response and the subsequent immune cell recruitment during inflammation.
Subject(s)
Colon/physiology , Endothelium, Vascular/physiology , Intercellular Adhesion Molecule-1/biosynthesis , Intestinal Mucosa/physiology , NF-kappa B/metabolism , Vascular Cell Adhesion Molecule-1/biosynthesis , Caco-2 Cells , Cell Communication , Cells, Cultured , Coculture Techniques , Colon/cytology , Endothelium, Vascular/cytology , Flow Cytometry , Humans , Intestinal Mucosa/cytology , Microcirculation/cytology , NF-kappa B/antagonists & inhibitorsSubject(s)
Isoantibodies/analysis , Lewis Blood Group Antigens , Antibody Specificity , Carbohydrate Sequence , Epitopes , HumansABSTRACT
A semi-automated system for determining the ABO group and Rh type of blood samples has been developed using a commercially available automated microplate (ELISA) reader and a microcomputer. Optimization of serologic, measurement and interpretation parameters was accomplished without significantly changing an existing manual procedure. The first pass noninterpretation rate of this system in the laboratory prior to field trials is 7.1%. A commercial system of this type should be cost-effective as a primary instrument for small to medium sized blood centers and transfusion services.
