ID: 183: Overcoming cell-to-cell heterogeneity reveals novel dynamic properties of IRF-3 and IFN-β

Ulfert Rand, David Spiller, Antony Adamson, Luka Cicin-Sain, Michael White

Research output: Contribution to journalArticle

Abstract

The type-I Interferon (IFN) response is an early innate immunity component and is antagonised by factors of a broad range of pathogenic viruses. The inherent dynamics of this cellular immune response are not well understood for two major reasons:
i.
Type-I IFN activation underlies temporal and quantitative cell-to-cell heterogeneity vastly restricting the validity of cell population-level studies.

ii.
Experimental stimulation of cellular receptors that lead to IFN activation is either achieved via viral infection or via endocytic uptake of synthetic compounds. Both approaches result in heterogeneity in response timing and response magnitude within the treated/infected cell population.


Using novel approaches comprising a dual reporter cell line (IRF-3-TagRFP, IFN-β-TurboGFP) and the electroporation of adherent cells together with extensive live-cell imaging, we identify dynamic properties of the central transcription factor IRF-3 at single-cell level upon stimulating the RIG-I receptor with synthetic dsRNA.

IRF-3 shows surprising dynamical properties controlling initial and subsequent IFN-β responses in single cells. IRF-3 contributes to the randomisation of IFN-β expression rates between cells upon initial stimulation. Furthermore, our reporter cell line visualises concentration-dependent effects of IRF-3 activation and IFN-β expression and the impact of TNF-α and IFN-β pre-treatment.

In summary, single-cell studies and defined intracellular receptor stimulation have unravelled response patterns and dynamic properties of the antiviral type-I IFN activation pathway that can now serve to characterise virus-activated responses in detail or design optimised vaccine adjuvants.
Original languageUndefined
Pages (from-to)98
Number of pages1
JournalCytokine
Volume76
Issue number1
DOIs
Publication statusPublished - 2015

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