Influence of endogenous pro-inflammatory cytokines on neutrophil-mediated damage of Candida albicans pseudohyphae, quantified in a modified tetrazolium dye assay.
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.
Vonk
Alieke G
AG
Department of Medicine, Division of General Internal Medicine, Radboud University Nijmegen Medical Center, The Netherlands.
Wieland
Catharina W
CW
Versteegen
Marieke
M
Verschueren
Ineke C
IC
Netea
Mihai G
MG
Joostent
Leo A B
LA
Verweij
Paul E
PE
Kullberg
Bart Jan
BJ
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*
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