title: A sensitive and specific assay for superoxide anion released by neutrophils or macrophages based on bioluminescence of polynoidin.
abstract: Using the luminescent protein polynoidin, present in the bioluminescent system isolated from the marine annelid Harmothoe lunulata, we have developed a new method to measure, specifically, superoxide anion (O2-) released by macrophages or neutrophils. A small quantity of an aqueous crude extract of polynoidin is used to detect O2- released by stimulated cells. Light emission is linearly dependent on the number of cells over a wide range (10(3) to 10(7) cells), and the assay is thus more sensitive than either luminol or ferricytochrome c reduction. Luminescence is enhanced 20% by mannitol, 80% by catalase, and is totally quenched by superoxide dismutase. For the same number of cells, neutrophils showed a threefold higher release of O2- and a twofold faster first-order light decay than stimulated macrophages, in accordance with data obtained by other methods.
authors.affiliation: Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.
authors.forename: P
authors.initials: P
authors.lastname: Colepicolo
authors.forename: V C
authors.initials: VC
authors.lastname: Camarero
authors.forename: M T
authors.initials: MT
authors.lastname: Nicolas
authors.forename: J M
authors.initials: JM
authors.lastname: Bassot
authors.forename: M L
authors.initials: ML
authors.lastname: Karnovsky
authors.forename: J W
authors.initials: JW
authors.lastname: Hastings
mesh_terms: D000818:Animals; D000845:Annelida / Q000032:analysis; D002374:Catalase / Q000494:pharmacology; D003574:Cytochrome c Group; D005260:Female; D005609:Free Radicals; D006168:Guinea Pigs; D008163:Luminescent Measurements; D008164:Luminescent Proteins / Q000302:isolation & purification; D008165:Luminol; D008264:Macrophages / Q000378:metabolism*; D008353:Mannitol / Q000494:pharmacology; D051379:Mice; D009504:Neutrophils / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000494:pharmacology; D013481:Superoxides / Q000032:analysis*
chemical_list: D003574:Cytochrome c Group; D005609:Free Radicals; D008164:Luminescent Proteins; D013481:Superoxides; D008353:Mannitol; D008165:Luminol; D002374:Catalase; D013482:Superoxide Dismutase