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Comparative Study
. 2014;163(2):154-62.
doi: 10.1159/000357036. Epub 2013 Dec 17.

Comparison of the interleukin-1β-inducing potency of allergenic spores from higher fungi (basidiomycetes) in a cryopreserved human whole blood system

Félix E Rivera-Mariani  1 , Kranthi Vysyaraju, Jesse Negherbon, Estelle Levetin, W Elliot Horner, Thomas Hartung, Patrick N Breysse
Affiliations
Comparative Study

Comparison of the interleukin-1β-inducing potency of allergenic spores from higher fungi (basidiomycetes) in a cryopreserved human whole blood system

Félix E Rivera-Mariani et al. Int Arch Allergy Immunol. 2014.

Abstract

Background: Spores from basidiomycete fungi (basidiospores) are highly prevalent in the atmosphere of urban and rural settings. Studies have confirmed their potential to affect human health as allergens. Less is known about their potential to serve as stimuli of the innate immune system and induce proinflammatory reactions.

Methods: In this study, we evaluated the proinflammatory potential of spores from 11 allergenic basidiomycete species (gilled: Pleurotus ostreatus,Oudemansiella radicata,Armillaria tabescens,Coprinus micaceus,Pluteus cervinus, and Chlorophyllum molybdites, and nongilled: Pisolithus arhizus,Merulius tremellosus,Calvatia cyathiformis,Lycoperdon pyriforme, andBoletus bicolor) based on their potency to induce the release of the proinflammatory cytokine interleukin (IL)-1β in a cryopreserved human whole blood system. In addition, the roles of morphological features of the spores (surface area, shape, and pigmentation) were examined for their role in the IL-1β-including potency of spores. Peripheral blood from healthy volunteers was collected, pooled, and cryopreserved. After stimulating the cryopreserved pooled blood with 10(6) to 10(3) basidiospores/ml, the concentration of IL-1β in culture supernatants was determined with ELISA.

Results: Basidiospores manifested concentration-dependent IL-1β-inducing potency, which was more marked among basidiospores from gilled basidiomycetes. At higher concentrations of basidiospores, the IL-1β-inducing potency could be differentiated in the cryopreserved human whole blood system. Morphological features did not correlate with the IL-1β-inducing potency of the basidiospores, suggesting that nonmorphological properties modulate the IL-1β-inducing potency.

Conclusion: Our data provide evidence of the proinflammatory potential of basidiospores, and the utility of cryopreserved human whole blood as a human-based in vitro system to study the immune reactivity of allergenic basidiospores.

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Conflict of interest statement

Disclosure of Conflict of Interest

The authors have no conflict of interest to disclose.

Figures

Figure 1
Figure 1
Images of basidiospores. (A) Pisolithus arizus, (B) Pleurotus ostreatus, (C) Oudemansiella radicata, (D) Armillaria tabescens, (E) Merulius tremullosus, (F) Calvatia cyathiformis, (G) Coprinus micaceus, (H) Lycoperdon pyriforme, (I) Pluteus cervinus, (J) Chlorophyllum molybdites, (K) Boletus bicolor. Images were acquired at 1000×.
Figure 2
Figure 2
Concentration-dependent responses of LPS and basidiospores in cryopreserved human whole blood. Cryopreserved human whole blood was stimulated with (A) LPS, (B) spores from gilled, (C) puffball and polypore basidiomycetes and the concentration of the proinflammatory cytokine IL-1β in culture supernatants determined with ELISA. Long-dash and dash lines represent the IL-1β-inducing potency of 5 and 2.5 EU/ml, respectively. Representative data from three independent experiments are shown. **p < 0.05 vs 5, 2.5 EU/ml of LPS; *p <0.05 vs 2.5 EU/ml of LPS.
Figure 3
Figure 3
Higher variability IL-1β-inducing potency at higher concentrations of basidiospores. Levene’s test of equal variances was performed to determine the concentration of basidiospores that, overall, induced the highest variability in the IL-1β-inducing after stimulation in the human whole blood. *p < 0.05 vs variance of 103, 104, 105 basidiospores/ml.
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