Comparison of inflammatory responses in human cells caused by lipopolysaccharides from Escherichia coli and from indigenous bacteria in aquatic environment

Abstract

The endotoxic activities of lipopolysaccharides (LPSs) in water samples are usually determined using a Limulus amoebocyte lysate (LAL) assay, but it is known that the determined activities do not always represent their inflammatory potency in humans. In this investigation, the inflammatory responses in three different human cells stimulated with Escherichia coli LPS, keratinocyte, CD14⁺ monocyte, and THP-1, were compared using cytokine secretion as biomarkers to develop novel in vitro assay systems for detecting changes in inflammatory potencies of endotoxins in aquatic environment. Only THP-1 with 6-h stimulation showed dose-dependent responses in the range of normal endotoxin levels in aquatic environment. Then, the inflammatory potency of environmental LPS, which was purified from river water, was tested using THP-1. The levels and patterns of cytokine secretion after the environmental LPS stimulation were completely different from E. coli LPS. Interleukin 8 (IL-8) secretions after the environmental LPS stimulation were approximately 10-fold higher than those after E. coli LPS stimulation. The environmental LPS also induced much higher levels of TNF-α secretions in THP-1. These results suggest that a diversity of LPS structures in aquatic environment could contribute to stronger and different inflammatory responses. This investigation indicated that the proposed THP-1 assay system could be useful for detecting the changes in inflammatory potencies caused by aquatic bacteria.

Keywords:

• Lipopolysaccharide (LPS)
• endotoxin
• human keratinocyte
• human blood cells
• cytokine secretion
• aquatic environment