title: Hypochlorite- and hypobromite-mediated radical formation and its role in cell lysis.
abstract: Activated leukocytes generate the potent oxidants HOCl and HOBr via the formation of H(2)O(2) and the release of peroxidase enzymes (myeloperoxidase, eosinophil peroxidase). HOCl and HOBr are potent microbiocidal agents, but excessive or misplaced production can cause tissue damage and cell lysis. In this study it is shown that HOBr induces red blood cell lysis at approximately 10-fold lower concentrations than HOCl, whereas with monocyte (THP1) and macrophage (J774) cells HOCl and HOBr induce lysis at similar concentrations. The role of radical formation during lysis has been investigated by EPR spin trapping, and it is shown that reaction of both oxidants with each cell type generates cell-derived radicals. Red blood cells exposed to nonlytic doses of HOCl generate novel nitrogen-centered radicals whose formation is GSH dependent. In contrast, HOBr gives rise to nitrogen-centered, membrane-derived protein radicals. With lytic doses of either oxidant, protein (probably hemoglobin)-derived, nitrogen-centered radicals are observed. Unlike the red blood cells, treatment of monocytes and macrophages with HOCl gives significant radical formation only under conditions where cell lysis occurs concurrently. These radicals are nitrogen-centered, cell-protein-derived species and have parameters identical to those detected with red blood cells and HOBr. Exposure of these cells to HOBr did not give detectable radicals. Overall these experiments demonstrate that HOCl and HOBr react with different selectivity with cellular targets, and that this can result in radical formation. This radical generation can precede, and may play a role in, cell lysis.
authors.affiliation: The Heart Research Institute, Camperdown, Sydney, New South Wales 2050, Australia.
authors.forename: C L
authors.initials: CL
authors.lastname: Hawkins
authors.forename: B E
authors.initials: BE
authors.lastname: Brown
authors.forename: M J
authors.initials: MJ
authors.lastname: Davies
mesh_terms: D001959:Bromates / Q000378:metabolism*; D002460:Cell Line; D004578:Electron Spin Resonance Spectroscopy; D004912:Erythrocytes / Q000378:metabolism; D005609:Free Radicals; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D006997:Hypochlorous Acid / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism; D009000:Monocytes / Q000378:metabolism; D009584:Nitrogen / Q000737:chemistry / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism; D011485:Protein Binding; D019097:Spin Trapping; D013997:Time Factors
chemical_list: D001959:Bromates; D005609:Free Radicals; D006997:Hypochlorous Acid; D006861:Hydrogen Peroxide; C027664:hypobromous acid; D009584:Nitrogen; D010100:Oxygen