Use of Indicator Cell Lines for Determining Inflammatory Gene Changes and Screening the Inflammatory Potential of Particulate and Non-Particulate Stimuli

Abstract

Ultrafine particulate matter, from environmental or industrial exposure, can induce the expression of inflammatory mediators and promote the production of reactive oxygen species (ROS), which can damage the alveolar epithelium of the lung. Previous studies have shown that various cellular stresses can activate signaling pathways that operate through the specific transcription factors (TF), AP-1, and nuclear factor (NF)-κ B that are known to regulate inflammatory gene expression. Persistent inflammation can induce a cascade of events that precedes the development of both acute and chronic fibrosis. From a murine Type II epithelial cell line, MLE15, a stable luciferase-transfected line, MLE15Luc2, was created. The luciferase reporter, operating under the guidance of a truncated human interleukin (IL)-8 promoter, contains NF-κ B and AP-1 DNA binding sites. MLE15Luc2 cells were exposed to inflammatory or particulate stimuli, of varying size fractions and composition, under standard culture conditions, and inflammatory gene transcription, represented by luciferase enzyme activity, was determined. Iκ Bα degradation appeared to be incongruent to changes in luciferase activity. The results were compared to those obtained using a stable luciferase-transfected human cell line, A549Luc1. Time-course data demonstrated increased luciferase enzyme activity, peaking by 6 h postexposure, and returning to baseline by 24 h, regardless of stimulus, in the absence of enhanced cytotoxicity. This suggests that key regulatory functions in these transfected cell lines are not clearly understood. These transfected cell lines may be useful for determining the inflammatory potential of various types of particulate and/or nonparticulate stimuli; however, conclusive signaling information cannot be gained from their use alone.