Cattle feedlot dust: Solubility in lung simulant fluid and stimulation of cytokine release from lung epithelial cells

Abstract

Beef cattle feed lots produce significant, local point source pollution of the atmosphere. The dusts generated in the CAFOs are complex mixture of fine and ultra fine particles, organic compounds, transition metals, and adsorbed toxic gases. Since each component is toxic in itself, we do not fully understand the relative importance of each component in the dust and their interactions to inducing inflammatory changes in the lung. We did extensive literature searches to understand the mechanism of dust toxicity in respiratory system. This lead to focusing on solubility of dust in lung simulant fluid, and in-vitro study of release of two common biomarkers of inflammatory processes IL-6 and IL-8 from lung epithelial cells. Various concentrations (1 to 50%) of the dust extract induced release of IL-6, and IL-8 from lung epithelial cell as indicators of pro-inflammatory changes (IL-6), and amplification and maintenance of inflammation (IL-8). IL-6 release had dose dependence; peak production was seen with 25% dust extract. IL-8 production went down as the concentration of the dust extract increased from 1% to 25%. However, 50% dust extract was cytotoxic to the cell leading to 10-15% cell viability. At non-cytotoxic concentrations for lung epithelial cells, production of IL-8 was reduced. These findings suggested that higher exposure concentration were required to initiate inflammation as indicated by IL-6 release. Lower exposure concentrations (1 and 5% extracts) were related to optimal release of IL-8 needed to amplify and maintain the inflammatory response. Inhibition of endotoxin didn't significantly change the pattern of IL-6 or IL-8 release from epithelial cells. This finding suggested that at least a portion of the mechanism by which particle induced cytokine release from the lung epithelial cells was not endotoxin dependent. Heating samples at 1200C for 5 minutes modified some of the toxic properties of the dust extracts but didn't completely detoxify it. We observed that longer incubation period was required to peak release for both IL-6 and IL-8. However, the higher concentration of sample (50% extract) found to be cytotoxic in non-heat treated sample was no longer cytotoxic and induced both IL-6 and IL-8 release from the lung epithelial cells. This result suggested that heat treatment could reduce some of the dust extract's cytotoxic properties. However, the extract's potential to induce peak cytokine release increased.