title: Detection of the halogenating activity of heme peroxidases in leukocytes by aminophenyl fluorescein.
abstract: The formation of hypochlorous and hypobromous acids by heme peroxidases is a key property of certain immune cells. These products are not only involved in defense against pathogenic microorganisms and in regulation of inflammatory processes, but contribute also to tissue damage in certain pathologies. After a short introduction about experimental approaches for the assessment of the halogenating activity in vitro and in cell suspensions, we are focusing on novel applications of fluorescent dye systems to detect the formation of hypochlorous acid (HOCl) in leukocytes. Special attention is directed to properties and applications of the non-fluorescent dye aminophenyl fluorescein that is converted by HOCl, HOBr, and other strong oxidants to fluorescein. This dye allows the detection of the halogenating activity in samples containing free myeloperoxidase and eosinophil peroxidase as well as in intact granulocytes using fluorescence spectroscopy and flow cytometry, respectively.
authors.affiliation: Institute for Medical Physics and Biophysics, University of Leipzig , Leipzig , Germany.
authors.forename: J
authors.initials: J
authors.lastname: Flemmig
authors.forename: J
authors.initials: J
authors.lastname: Remmler
authors.forename: J
authors.initials: J
authors.lastname: Zschaler
authors.forename: J
authors.initials: J
authors.lastname: Arnhold
mesh_terms: D000814:Aniline Compounds / Q000737:chemistry*; D000818:Animals; D047209:Eosinophil Peroxidase / Q000032:analysis / Q000378:metabolism*; D005434:Flow Cytometry; D005452:Fluoresceins / Q000737:chemistry*; D005456:Fluorescent Dyes; D054879:Halogenation; D006801:Humans; D006997:Hypochlorous Acid / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D007962:Leukocytes / Q000201:enzymology*; D009195:Peroxidase / Q000032:analysis / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: D000814:Aniline Compounds; D005452:Fluoresceins; D005456:Fluorescent Dyes; C508569:aminophenyl fluorescein; D006997:Hypochlorous Acid; D047209:Eosinophil Peroxidase; D009195:Peroxidase
keywords: aminophenyl fluorescein; eosinophil peroxidase; eosinophils; hypobromous acid; hypochlorous acid; myeloperoxidase; neutrophils