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IFN-β Expression Is Directly Activated in Human Neutrophils Transfected with Plasmid DNA and Is Further Increased via TLR-4–Mediated Signaling

Abstract : Upon LPS binding, TLR4 activates a MyD88-dependent pathway leading to the transcriptional activation of proinflammatory genes, as well as a MyD88-independent/TRIF-dependent pathway, responsible for the transcriptional induction of IFN-b. Previous findings delineated that human neutrophils are unable to induce the transcription of IFN-b in response to TLR4 stimulation. Because neutrophils do not express protein kinase C «, a molecule recently reported as essential for initiating the MyD88independent/TRIF-dependent pathway, we optimized an electroporation method to transfect PKC« into neutrophils with very high efficiency. By doing so, a significant IFN-b mRNA expression was induced, in the absence of LPS stimulation, not only in PKC«-overexpressing neutrophils but also in cells transfected with a series of empty DNA plasmids; however, LPS further upregulated the IFN-b transcript levels in plasmid-transfected neutrophils, regardless of PKC« overexpression. Phosphoimmunoblotting studies, as well as chromatin immunoprecipitation assays targeting the IFN-b promoter, revealed that IFN-b mRNA induction occurred through the cooperative action of IRF3, activated by transfected DNA, and NF-kB, activated by LPS. Additional immunoblotting and coimmunoprecipitation studies revealed that neutrophils constitutively express various cytosolic DNA sensors, including IFN-inducible protein 16, leucine-rich repeat (in Flightless I) interacting protein-1, and DDX41, as well as that IFN-inducible protein 16 is the intracellular receptor recognizing transfected DNA. Consistently, infection of neutrophils with intracellular pathogens, such as Bartonella henselae, Listeria monocytogenes, Legionella pneumophila, or adenovirus type 5, promoted a marked induction of IFN-b mRNA expression. Taken together, these data raise questions about the role of PKC« in driving the MyD88-independent/TRIF-dependent response and indicate that human neutrophils are able to recognize and respond to microbial cytosolic DNA.
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https://hal.uvsq.fr/hal-03712461
Contributor : Vincent LE MOIGNE Connect in order to contact the contributor
Submitted on : Tuesday, July 5, 2022 - 10:23:52 PM
Last modification on : Tuesday, July 5, 2022 - 10:23:52 PM

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Nicola Tamassia, Flavia Bazzoni, Vincent Le Moigne, Federica Calzetti, Caterina Masala, et al.. IFN-β Expression Is Directly Activated in Human Neutrophils Transfected with Plasmid DNA and Is Further Increased via TLR-4–Mediated Signaling. Journal of Immunology, 2012, 189 (3), pp.1500 - 1509. ⟨10.4049/jimmunol.1102985⟩. ⟨hal-03712461⟩

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