Abstract
SUMMARYMacrophages initiate inflammatory responses via the transcription factor NFκB. The temporal pattern of NFκB activity determines which genes are expressed and thus, the type of response that ensues. Here, we examined how information about the stimulus is encoded in the dynamics of NFκB activity. We generated an mVenus-RelA reporter mouse line to enable high-throughput live-cell analysis of primary macrophages responding to host- and pathogen-derived stimuli. An information-theoretic workflow identified six dynamical features—termed signaling codons—that convey stimulus information to the nucleus. In particular, oscillatory trajectories were a hallmark of responses to cytokine but not pathogen-derived stimuli. Single-cell imaging and RNA sequencing of macrophages from a mouse model of Sjögren’s syndrome revealed inappropriate responses to stimuli, suggestive of confusion of two NFκB signaling codons. Thus, the dynamics of NFκB signaling classify immune threats through six signaling codons, and signal confusion based on defective codon deployment may underlie the etiology of some inflammatory diseases.
Highlights
Autoimmune pathologies are characterized by the presence of auto-antibodies and immune attack of specific tissues, but the etiology is not uniform (Marshak-Rothstein, 2006)
Macrophages, differentiated from primary bone-marrow cells derived from homozygous Relav/v mice, showed normal levels of nuclear NFkB binding activity (Figure S1C)
Upon stimulation with a variety of different ligands and doses, and time-lapse imaging over 21 h (Figure 1B), the amount of nuclear NFkB fluorescence was quantitated in single cells using a fully automated imageprocessing pipeline that enabled tracking of live cells using minimal levels of a nuclear marker (Selimkhanov et al, 2014; Zambrano et al, 2016) and label-free identification and segmentation of cell cytoplasm
Summary
Autoimmune pathologies are characterized by the presence of auto-antibodies and immune attack of specific tissues, but the etiology is not uniform (Marshak-Rothstein, 2006). Sjogren’s syndrome (SS) is a systemic autoimmune disorder that is characterized by progressive destruction of tissues exposed to the environment, such as eye, mouth and throat, and skin rashes (Marshak-Rothstein, 2006). Several genetic variants in regulators of the transcription factor NFkB are associated with SS patients (Lisi et al, 2012; Nordmark et al, 2013; Ou et al, 2008; Sisto et al, 2013), and a mouse strain containing similar variants recapitulates some of the SS pathognomonic characteristics (Peng et al, 2010). It remains unknown how these alleles affect NFkB dynamics
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