Cellular reporter for inflammation.

Abstract

Inflammatory response in cells are often measured through expression of a key transcription factor called NF-B. Currently, there are commercial green fluorescent protein (GFP) assays available for in vitro cell lines only. There are no in vivo systems that can report NF-B activation in hematopoietic stem cells. As such, this project is aimed at trying to develop such an in vivo assay, albeit at a preliminary stage. We have a retroviral reporter for NF-B in which we conducted experiments to test the workability of this reporter. Through a process called retroviral-mediated gene transfer, we transduced bone marrow cells with the reporter and tested for GFP expression in presence of pro-inflammatory stimuli. We had found that our reporter could clearly show GFP expression well when the transduced bone marrow cells were induced to undergo inflammation. However, upon differentiating the transduced bone marrow cells into macrophage and stimulating to induce inflammation, GFP expression was low which we suspect was due to a silencing of the reporter during differentiation. Further experiments need to be done in vivo to confirm whether the reporter is able to express GFP in presence of pro-inflammatory stimuli.