Measurement of IL-6 levels in live cells using RNA detection probe with Imaging Flow Cytometer

Abstract

Abstract Acute inflammatory responses to infection and tissue damage are orchestrated by release of cytokines, primarily from macrophages and mast cells, but also by endothelium and tumor cells. A key initiator of the inflammatory response is tumor necrosis factor alpha (TNF-alpha), which binds to its receptor on target cells and activates a pro-inflammatory transcriptional program via the transcription factor NFkB. A critical downstream target of NFkB is the gene encoding interleukin-6, which stimulates an array of anti-infection processes, including synthesis of acute-phase proteins by the liver, proliferation of B cells, neutrophil production in bone marrow, and differentiation of Th17 helper T cells. Although beneficial in the context of attacking infection, dysregulation of IL-6 plays a deleterious role in autoimmune disease and cancer. In this study, we introduce a new method for measuring NFkB translocation by looking at the increase in IL-6 mRNA levels in the THP-1 suspension cell line using SmartFlare™ RNA detection probes. The ability to detect RNA in live cells and to acquire multi-spectral images of large numbers of cells allows for the accurate assessment of IL-6 mRNA levels in THP-1 cells.