1. Influence of endogenous pro-inflammatory cytokines on neutrophil-mediated damage of Candida albicans pseudohyphae, quantified in a modified tetrazolium dye assay.
- Author
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Vonk AG, Wieland CW, Versteegen M, Verschueren IC, Netea MG, Joostent LA, Verweij PE, and Kullberg BJ
- Subjects
- Animals, Hyphae immunology, Indicators and Reagents, Interferon-gamma deficiency, Interleukins deficiency, Lymphotoxin-alpha deficiency, Mice, Mice, Inbred C57BL, Mice, Knockout, Specific Pathogen-Free Organisms, Statistics, Nonparametric, Tetrazolium Salts, Tumor Necrosis Factor-alpha deficiency, Candida albicans immunology, Interferon-gamma immunology, Interleukins immunology, Lymphotoxin-alpha immunology, Neutrophils immunology, Tumor Necrosis Factor-alpha immunology
- 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.
- Published
- 2005
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