title: Regulation of CD59 expression on K562 cells: effects of phorbol myristate acetate, cross-linking antibody and non-lethal complement attack.
abstract: CD59 is the major membrane attack complex of complement (MAC) inhibiting protein on human cells. Its regulation is therefore an important factor in determining the fate of cells at sites of complement activation. We have chosen the K562 erythroleukaemia cell line as a model for studies of the regulation of CD59 expression, because it has previously been reported that phorbol 12-myristate 13-acetate (PMA) caused a 15-fold up-regulation of CD59 mRNA in these cells, implying a substantial capacity for CD59 synthesis. However, no assessment of CD59 protein expression was made in these studies. We show here that surface expression of CD59, as assessed by flow cytometry, was increased four-fold over a 16-hr incubation with PMA, whereas surface expression of decay-accelerating factor (DAF) (CD55) and membrane cofactor protein (MCP) (CD46) was not altered. The newly expressed CD59 was functionally active and anchored through glycosyl-phosphatidylinositol (GPI). Increased expression was dependent upon de novo protein synthesis. CD59 released into cell supernatant was also increased seven-fold by PMA, this 'secreted' CD59 retained its GPI anchor. Non-lethal complement attack did not alter CD59 expression but antibody cross-linking of CD59 caused a rapid loss of the CD59-antibody complexes. However, CD59 was quickly restored to pre-attack levels. This rapid restoration was not dependent upon protein synthesis, suggesting release from preformed stores.
authors.affiliation: Department of Medical Biochemistry, University of Wales College of Medicine, Cardiff, UK.
authors.forename: K J
authors.initials: KJ
authors.lastname: Marchbank
authors.forename: B P
authors.initials: BP
authors.lastname: Morgan
authors.forename: C W
authors.initials: CW
authors.lastname: van den Berg
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D015153:Blotting, Western; D018957:CD59 Antigens; D003169:Complement Inactivator Proteins / Q000378:metabolism*; D003165:Complement System Proteins / Q000276:immunology*; D003513:Cycloheximide / Q000494:pharmacology; D003602:Cytotoxicity, Immunologic; D003609:Dactinomycin / Q000494:pharmacology; D006801:Humans; D007942:Leukemia, Experimental / Q000276:immunology; D008562:Membrane Glycoproteins / Q000378:metabolism*; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010727:Phosphoric Diester Hydrolases / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000187:drug effects / Q000276:immunology*
chemical_list: D015703:Antigens, CD; D018957:CD59 Antigens; D003169:Complement Inactivator Proteins; D008562:Membrane Glycoproteins; D003609:Dactinomycin; D003165:Complement System Proteins; D003513:Cycloheximide; D010727:Phosphoric Diester Hydrolases; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D013755:Tetradecanoylphorbol Acetate