Role of cytoskeletal elements in expression of monocyte urokinase plasminogen activator receptor, activation-associated antigen Mo3.
Peripheral blood monocytes exposed to bacterial products, phorbol esters, cyclic AMP, and cyclic AMP analogs express cell surface activation protein Mo3, which is the human urokinase plasminogen activator receptor (uPA-R). uPA-R is expressed by circulating monocytes from patients with multiple sclerosis (MS). We examined the role of cytoskeletal elements in the surface expression and subcellular distribution of uPA-R in nonactivated and lipopolysaccharide-activated monocytes and in monocytes from patients with MS. By using immunofluorescence techniques and confocal laser microscopy, we found that in unactivated monocytes, cytoplasmic uPA-R is found to one side of the nucleus, colocalizing with the Golgi. Upon activation with lipopolysaccharide, cytoplasmic Mo3-uPA-R becomes dispersed throughout the cytoplasm and projections concomitant with an increase in the monocyte perimeter (spreading). Cytoplasmic dispersion, as well as cell surface deposition, is dependent on microtubule integrity. Cell surface deposition of uPA-R upon activation is reduced by colchicine, which disrupts microtubules; however, once associated at the cell surface, uPA-R becomes associated with microfilaments via vinculin. Disruption of microfilaments with cytochalasin also alters surface expression of immunologically reactive uPA-R, as well as the distribution pattern. Monocytes from patients with MS display the uPA-R distribution pattern characteristic of an activated monocyte.
Washington
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Department of Neurology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
Dore-Duffy
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D008841:Actin Cytoskeleton / Q000201:enzymology; D000941:Antigens / Q000276:immunology; D002478:Cells, Cultured / Q000201:enzymology / Q000276:immunology; D003599:Cytoskeleton / Q000201:enzymology* / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008070:Lipopolysaccharides; D018613:Microscopy, Confocal; D008870:Microtubules / Q000201:enzymology; D009000:Monocytes / Q000201:enzymology* / Q000276:immunology; D009103:Multiple Sclerosis / Q000201:enzymology / Q000276:immunology; D014568:Urokinase-Type Plasminogen Activator / Q000276:immunology*
D000941:Antigens; D008070:Lipopolysaccharides; D014568:Urokinase-Type Plasminogen Activator