Determination of inflammatory responses to Aspergillus versicolor and endotoxin with human cryo-preserved blood as a suitable tool

The aim of this study was to characterize the mediator release in the whole blood assay induced by stimulation with an extract of the indoor mold A. versicolor. In addition, the effect of concomitant stimulation with E. coli endotoxin and A. versicolor, and the involvement of the relevant Toll-like receptors were investigated. The stimulation of cryo-preserved whole blood with 400pg/ml E. coli endotoxin induced a significant IL-1β release (976±133pg/ml), whereas, none of the tested A. versicolor concentrations (10–1000mg/ml) caused IL-1β release. However, stimulation with A. versicolor resulted in a maximum IL-8 release of 6200pg/ml. The simultaneous stimulation with 5pg/ml E. coli endotoxin and A. versicolor induced a synergistic effect on both IL-1β and IL-8 release. Addition of anti-TLR-4 reduced the release of IL-1β and IL-8 up to 56% and 43%, respectively. In summary, the whole blood assay is useful to characterize the inflammatory potential of A. versicolor, especially when the release of the pro-inflammatory chemokine IL-8 is included. Additionally, it is also possible to study the influence of cell-surface receptors of the innate immunity involved in the effects of specific microbial stimuli.