Effects of Antifungal Agents on the Production of Cytokines from Macrophages Stimulated by Aspergillus fumigatus conidia

ABSTRACT

BACKGROUND: We wanted to evaluate the immunomodulatory effect of three antifungal agents, amphotericin B (AmB), micafungin (MF), itraconazole (ITZ), in the aspect of cytokine production and expression of nuclear factor-kappaB (NF-kappaB) in the Aspergillus fumigatus conidia treated-RAW264.7 cells, a murine alveolar macrophage cell line. MATERIALS AND METHODS: We evaluated cytotoxic effect of antifungal agents using commercial cell proliferation assay. Tumor necrosis factor (TNF)-alpha and interleukin (IL)-10 production according to stimulation (control, A. fumigatus conidia only, conidia + antifungal drug, conidia + antifungal drug + G(M)-CSF) was evaluated and compared each using commercial ELISA method. NF-kappaB activation was evaluated by western blot analysis. RESULTS: AmB, MF, ITZ showed dose-dependent cytotoxic effect on the tested cells. Stimulation of cells by A. fumigatus conidia induced TNF-alpha production. Pretreatment of AmB at concentration not affecting cellular survival did not change the production of TNF-alpha compared to conidia treated cells, but pretreament of MF or ITZ showed reduced amount of TNF-alpha production compared to conidia treated cells. AmB also showed synergistic effect on TNF-alpha production when simultaneously treated with GM-CSF. IL-10 production was markedly increased when the cells were treated with AmB with conidia. MF and ITZ induced less increase of IL-10 production than AmB. AmB also showed synergistic effect on the production of IL-10 when treated with GM-CSF simultaneously. A. fumigatus conidia enhanced expression of NF-kappaB. The degree of NF-kappaB expression was associated with the amount of TNF-alpha and IL-10 produced. CONCLUSION: The antifungal agents we used in this experiment showed that decreased TNF-alpha production and increased IL-10 production from the RAW264.7 cells stimulated by A. fumigatus conidia after pretreatment of antifungal agents. But more studies, such as the association between immunomodulatory effect, antifungal activity and difference of signal pathway of cellular activation according to drugs should be performed.