title: Influence of endogenous pro-inflammatory cytokines on neutrophil-mediated damage of Candida albicans pseudohyphae, quantified in a modified tetrazolium dye assay.
abstract: For quantitative assessment of polymorphonuclear granulocyte (PMN)-mediated pseudohyphal damage, an improved tetrazolium (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide; XTT) dye assay was developed. The modified assay proved to be a reliable indicator of viable pseudohyphal inoculum sizes. In addition, the influence of various endogenous pro-inflammatory cytokines on the capacity of PMN to damage Candida albicans pseudohyphae was investigated. PMN obtained from mice in which the genes encoding for tumor necrosis factor-alpha/lymphotoxin-alpha (TNF/LT), interferon-gamma (IFNgamma), interleukin (IL)-1alpha, or IL-1beta were disrupted, showed a significantly reduced pseudohyphal damage capacity in comparison with control PMN. The reduction amounted 25% for TNF-/- LT-/-, 11% for IFNgamma-/-, 21% for IL-1alpha-/-, and 34% for IL-1alpha-/-beta-/- PMN. In contrast, deficiency of IL-12 or IL-18 did not result in a diminished capacity to damage pseudohyphae and the capacity of PMN to damage Candida pseudohyphae was even slightly increased by 10% in IL-18-/- mice. These data suggest that endogenous pro-inflammatory cytokines are able to modulate antihyphal activity of PMN, the main effector cells against disseminated candidiasis by virtue of their capacity to kill both Candida blastoconidia and pseudohyphae.
authors.affiliation: Department of Medicine, Division of General Internal Medicine, Radboud University Nijmegen Medical Center, The Netherlands.
authors.forename: Alieke G
authors.initials: AG
authors.lastname: Vonk
authors.forename: Catharina W
authors.initials: CW
authors.lastname: Wieland
authors.forename: Marieke
authors.initials: M
authors.lastname: Versteegen
authors.forename: Ineke C
authors.initials: IC
authors.lastname: Verschueren
authors.forename: Mihai G
authors.initials: MG
authors.lastname: Netea
authors.forename: Leo A B
authors.initials: LA
authors.lastname: Joostent
authors.forename: Paul E
authors.initials: PE
authors.lastname: Verweij
authors.forename: Bart Jan
authors.initials: BJ
authors.lastname: Kullberg
mesh_terms: D000818:Animals; D002176:Candida albicans / Q000276:immunology*; D025301:Hyphae / Q000276:immunology; D007202:Indicators and Reagents; D007371:Interferon-gamma / Q000172:deficiency / Q000276:immunology*; D007378:Interleukins / Q000172:deficiency / Q000276:immunology*; D008233:Lymphotoxin-alpha / Q000172:deficiency / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009504:Neutrophils / Q000276:immunology*; D013047:Specific Pathogen-Free Organisms; D018709:Statistics, Nonparametric; D013778:Tetrazolium Salts; D014409:Tumor Necrosis Factor-alpha / Q000172:deficiency / Q000276:immunology*
chemical_list: D007202:Indicators and Reagents; D007378:Interleukins; D008233:Lymphotoxin-alpha; D013778:Tetrazolium Salts; D014409:Tumor Necrosis Factor-alpha; C059087:2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide; D007371:Interferon-gamma