Human inflammatory response of endotoxin affected by particulate matter-bound transition metals.

Abstract Bacterial endotoxins are a component of particulate matter (PM) with anticipated health implications, yet we know little about how host reception of endotoxin through toll-like receptor 4 (TLR4) is affected by its association with other PM components. Subsequently, we investigated the relationship between endotoxin concentration (recombinant Factor C (rFC) assay) and host recognition (HEK Blue-TLR4 NF-kB reporter cell line based assay) in various compositions of urban PM, including road traffic, industrial and urban green land use classes. While the assays did not correlate strongly between each other, the TLR4 reporter cell line was found to be better correlated to the IL-8 response of PM. Furthermore, the ability of the quantified endotoxin (rFC assay) to stimulate the TLR4/MD-2 complex was significantly affected by the urban land use class, where traffic locations were found to be significantly higher in bioactive endotoxin than the industrial and green locations. We subsequently turned our attention to PM composition and characterized the samples based on transition metal content (through ICP-MS). The effect of nickel and cobalt – previously reported to activate the hTLR4/MD-2 complex – was found to be negligible in comparison to that of iron. Here, the addition of iron as a factor significantly improved the regression model between the two endotoxin assays, explaining 77% of the variation of the TLR4 stimulation and excluding the significant effect of land use class. Moreover, the effect of iron proved to be more than a correlation, since dosing LPS with Fe²⁺ led to an increase up to 64% in TLR4 stimulation, while Fe²⁺ without LPS was unable to stimulate a response. This study shows that endotoxin quantification assays (such as the rFC assay) may not always correspond to human biological recognition of endotoxin in urban PM, while its toxicity can be synergistically influenced by the associated PM composition. Graphical abstract Image 1 Highlights • Quantified endotoxin not strongly correlated to host recognition through hTLR4. • HEK TLR4 cell line better assay for IL8 response than rFC-quantified endotoxin. • Bioactivity of atmospheric endotoxin influenced by land-use class. • Ferrous iron modulates host endotoxin response. Although endotoxins are typically quantified in ambient air, its toxicity is significantly influenced by the associated PM composition (e.g. as shown in this study with iron, one of the most abundant transition metals found in urban PM). [ABSTRACT FROM AUTHOR]