Examining the response of primary Human Bronchial Epithelial Cells in an Air liquid interface culture system after stimulation with concentrated Exhaled Breath Condensate

Introduction: Exhaled breath condensate (EBC) is a noninvasive method to analyze inflammatory mediators present in the lower respiratory system. EBC contains aerosolized particles reflecting the airway lining fluid with variations of mediators from different lung diseases. However there are limitations to EBC measurements due to the low concentration of proteins and biomarkers. In different lung diseases, such as alpha-1 antitrypsin deficiency (AATD), chronic obstructive pulmonary disease (COPD) and asthma, proteins and cytokines represented in air liquid interface (ALI) system after EBC stimulation are expected to be different.

Objective: The objective of this study is the implementation of a method to analyze the different contents of low concentrated proteins and cytokines in EBC.

Methods: The approach is the concentration of EBC with differing columns, and the subsequent stimulation of differentiated primary human bronchial epithelial cells (pHBECs) in the ALI system with concentrated EBC as an airway epithelial lining fluid. Accordingly, we analyze the inflammatory response of the HBEC in the ALI reflected in the regulation of Interleukin 6 (IL-6) and Monocyte Colony stimulating Protein (MCP) as a pilot approach. Following incubation we expect to see cell transcription profile responses as well as cytokine secretion in the culture medium.

Result: Preliminary results in the first experiment with stimulation of unconcentrated EBC suggest measurable differences in MCP-1and IL-6 regulation between healthy controls, smokers and asthma patients.

Conclusion: Response examination of the pHBEC after stimuli with concentrated EBC may contribute to the understanding of the pathophysiology in lung diseases. The non invasive approach combined with a cell culture model offers new opportunities for future experiments.