An experimental model for the determination of immunomodulating effects by volatile compounds

An in vitro exposure system was developed to enable simultaneous exposure of primary cells or cell lines to defined concentrations of volatile organic compounds (VOC) without the necessity of a constant-flow exposure system. Toluene was used as model VOC and administered via the gas phase. CD3/CD28-stimulated human peripheral blood mononuclear cells (PBMC) were used as indicator cells. Vitality/proliferation of PBMC was tested using the MTT assay and their functional reactivity using cytokine ELISA for interferon-gamma (IFN-gamma), interleukin-4 (IL-4), IL-13, and tumor-necrosis-factor-alpha (TNF-alpha). Chemical analysis using headspace gas chromatography confirmed that this new method guaranties reproducible VOC exposure (R2 = 0.995 for the correlation between external toluene concentration and toluene in the cell culture). While cytotoxic effects were not observed, dose-dependent toluene effects on functional reactivity of PBMC were found. The secretion of IFN-gamma, IL-4, and IL-13 was inhibited at concentrations of 72.5 g/m3 and above, whereas the TNF-alpha production was increased. Since the presented in vitro model ensures toluene exposure in concentrations comparable to the real situation, and allows the investigation of dose-dependent immunomodulatory toluene effects in concentrations without cytotoxicity, this method first described here is introduced as useful tool in analysis of VOC-triggered effects on immune cells.