title: Endothelial cells potentiate oxidant-mediated Kupffer cell phagocytic killing.
abstract: Phagocytosis and killing of circulating organisms by Kupffer cells (KCs) are discrete, important components of host defense. However, the killing mechanism(s) are not fully understood, and the potential role of adjacent nonparenchymal cells such as hepatic endothelial cells has not been defined. Rat KCs -/+ an hepatic endothelial cell enriched cellular fraction (HECEF) were incubated with Candida parapsilosis and assayed for phagocytosis and phagocytic killing by validated fluorochromatic vital staining. The role of reactive oxygen metabolites in KC phagocytic functions was examined by inhibition with superoxide dismutase and/or catalase. Diphenyleneiodonium and allopurinol were used to examine the potential roles of NADPH oxidase and xanthine oxidase, respectively, in generating these toxic oxidants. Coculture with HECEF increased KC phagocytic activity (from 75% to 88%) and candidacidal activity (from 20% to 31%). Superoxide dismutase, catalase, diphenyleneiodonium, or allopurinol caused inhibition of candidacidal activity, but did not affect phagocytosis, and did not block the potentiation of phagocytosis or of killing caused by coculture with HECEF. Reactive oxygen intermediates generated by both NADPH oxidase and xanthine oxidase-dependent pathways are important in KC killing of Candida parapsilosis. In vitro, KC phagocytosis and killing are potentiated (via a non-oxidant-mediated mechanism) by coculture with a preparation of hepatic non-parenchymal cells composed primarily of endothelial cells.
authors.affiliation: Department of Surgery, Johns Hopkins University School of Medicine, Baltimore, MD 21287-8611, USA.
authors.forename: D A
authors.initials: DA
authors.lastname: Potoka
authors.forename: S
authors.initials: S
authors.lastname: Takao
authors.forename: T
authors.initials: T
authors.lastname: Owaki
authors.forename: G B
authors.initials: GB
authors.lastname: Bulkley
authors.forename: A S
authors.initials: AS
authors.lastname: Klein
mesh_terms: D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D002175:Candida / Q000276:immunology; D002374:Catalase / Q000494:pharmacology; D018920:Coculture Techniques; D004727:Endothelium / Q000166:cytology / Q000502:physiology; D005609:Free Radicals / Q000378:metabolism; D007728:Kupffer Cells / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D008297:Male; D019255:NADPH Oxidases / Q000378:metabolism; D009861:Onium Compounds / Q000494:pharmacology; D016877:Oxidants / Q000378:metabolism*; D010587:Phagocytosis / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D011917:Rats, Inbred Lew; D017382:Reactive Oxygen Species / Q000378:metabolism; D013482:Superoxide Dismutase / Q000494:pharmacology; D014969:Xanthine Oxidase / Q000378:metabolism
chemical_list: D005609:Free Radicals; D009861:Onium Compounds; D016877:Oxidants; D017382:Reactive Oxygen Species; D000493:Allopurinol; C007517:diphenyleneiodonium; D002374:Catalase; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase; D019255:NADPH Oxidases