A sensitive detection of neutrophil activation by fluorescence quenching of membrane inserted singlet oxygen probe.
Fluorescence emission intensity of 1,3-diphenylisobenzofuran incorporated in polymorphonuclear granulocytes plasma membranes was investigated in basal conditions and during stimulation with different activators. Phorbol myristate acetate, known as the most effective "oxygen burst" inducer, produced a larger decrease in 1,3-diphenylisobenzofuran fluorescence intensity than 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine (platelet activating factor) and N-formyl-methionyl-leucyl-phenylalanine, known as weak stimulants of oxygen uptake. Diphenyl iodonium an inhibitor of leukocyte NADPH oxidase, and the singlet oxygen selective trap alpha-terpinene inhibited the quenching effect of phorbol myristate acetate on 1,3-diphenylisobenzofuran fluorescence. These data suggest formation of singlet oxygen in activated leukocytes and demonstrate that measurement of 1,3-diphenylisobenzofuran fluorescence intensity provides a new sensitive method of detection of neutrophils activation.
Tanfani
F
F
Institute of Biochemistry, Medical School, University of Ancona, Italy.
Fiorini
R
R
Tartaglini
E
E
Kantar
A
A
Wozniak
M
M
Antosiewicz
J
J
Bertoli
E
E
D001572:Benzofurans / Q000737:chemistry*; D001713:Biphenyl Compounds / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D000081005:Cyclohexane Monoterpenes; D017665:Hydroxyl Radical / Q000378:metabolism; D039821:Monoterpenes; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009504:Neutrophils / Q000378:metabolism*; D009861:Onium Compounds / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism*; D010777:Photochemistry; D010972:Platelet Activating Factor / Q000494:pharmacology; D026082:Singlet Oxygen; D013050:Spectrometry, Fluorescence; D013729:Terpenes / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D001572:Benzofurans; D001713:Biphenyl Compounds; D000081005:Cyclohexane Monoterpenes; D039821:Monoterpenes; D009861:Onium Compounds; D010972:Platelet Activating Factor; D013729:Terpenes; C031291:diphenyliodonium; D026082:Singlet Oxygen; D017665:Hydroxyl Radical; C018669:gamma-terpinene; C011238:1,3-diphenylisobenzofuran; D009240:N-Formylmethionine Leucyl-Phenylalanine; D013755:Tetradecanoylphorbol Acetate; D010100:Oxygen