Expression of adhesion molecules is specific and time-dependent in cytokine-stimulated endothelial cells in culture.
The time course of expression of the adhesion molecules E-selectin, VCAM-1, ICAM-1 and PECAM-1 was studied in interleukin-1 beta-stimulated human umbilical vein cells (HUVEC) and the subcellular sites of synthesis were determined by means of fluorescence immunohistochemistry. The maximal number of cells labelled for E-selectin was observed at 2-4 h, for VCAM-1 at 4-8 h and ICAM-1 at 6-72 h. At 8 h, E-selectin and VCAM-1 started to disappear, but ICAM-1-positive cells persisted. PECAM-1 was constitutively expressed. De novo synthesis for E-selectin started at 1 h and for both, VCAM-1 and ICAM-1 at 1.5-2 h. Maximal synthetic activity was observed at 2.5-4 h for E-selectin and at 4-6 h for VCAM-1 and ICAM-1; thereafter, synthesis slowly decreased. Transport granules occurred at 1.5 h for E-selectin and 4 h for VCAM-1; they were absent for ICAM-1. Diffuse cellular and membrane labelling indicative of the functional activity of the adhesion molecules began at 2-4 h for E-selectin, and 4 h for VCAM, but was constitutively present for ICAM-1. In conclusion, each adhesion molecule shows a specific time-dependent course of appearance and disappearance in interleukin-1 beta-stimulated HUVECs in accordance with their physiological role in vivo. These morphological results confirm data obtained by flow cytometry and Western blotting, but they provide new information about the behaviour of individual cells with regard to the sites of synthesis and cellular localization of the adhesion molecules.
Scholz
D
D
Max-Planck-Institute, Department of Experimental Cardiology, Bad Nauheim, Germany. dscholz@kerckhoff.mpg.de
Devaux
B
B
Hirche
A
A
Pötzsch
B
B
Kropp
B
B
Schaper
W
W
Schaper
J
J
D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis / Q000096:biosynthesis* / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000096:biosynthesis* / Q000187:drug effects; D002478:Cells, Cultured / Q000737:chemistry / Q000166:cytology / Q000187:drug effects; D016207:Cytokines / Q000494:pharmacology*; D019040:E-Selectin / Q000032:analysis / Q000096:biosynthesis / Q000187:drug effects; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D018799:Intercellular Adhesion Molecule-1 / Q000032:analysis / Q000096:biosynthesis / Q000187:drug effects; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology; D019010:Vascular Cell Adhesion Molecule-1 / Q000032:analysis / Q000096:biosynthesis / Q000187:drug effects
D015214:Antigens, Differentiation, Myelomonocytic; D015815:Cell Adhesion Molecules; D016207:Cytokines; D019040:E-Selectin; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1