title: Early innate recognition of herpes simplex virus in human primary macrophages is mediated via the MDA5/MAVS-dependent and MDA5/MAVS/RNA polymerase III-independent pathways.
abstract: Innate recognition of viruses is mediated by pattern recognition receptors (PRRs) triggering expression of antiviral interferons (IFNs) and proinflammatory cytokines. In mice, Toll-like receptor 2 (TLR2) and TLR9 as well as intracellular nucleotide-sensing pathways have been shown to recognize herpes simplex virus (HSV). Here, we describe how human primary macrophages recognize early HSV infection via intracellular pathways. A number of inflammatory cytokines, IFNs, and IFN-stimulated genes were upregulated after HSV infection. We show that early recognition of HSV and induction of IFNs and inflammatory cytokines are independent of TLR2 and TLR9, since inhibition of TLR2 using TLR2 neutralizing antibodies did not affect virus-induced responses and the macrophages were unresponsive to TLR9 stimulation. Instead, HSV recognition involves intracellular recognition systems, since induction of tumor necrosis factor alpha (TNF-α) and IFNs was dependent on virus entry and replication. Importantly, expression of IFNs was strongly inhibited by small interfering RNA (siRNA) knockdown of MAVS, but this MAVS-dependent IFN induction occurred independently of the recently discovered polymerase III (Pol III)/RIG-I DNA sensing system. In contrast, induction of TNF-α was largely independent of MAVS, suggesting that induction of inflammatory cytokines during HSV infection proceeds via a novel pathway. Transfection with ODN2006, a broad inhibitor of intracellular nucleotide recognition, revealed that nucleotide-sensing systems are employed to induce both IFNs and TNF-α. Finally, using siRNA knockdown, we found that MDA5, but not RIG-I, was the primary mediator of HSV recognition. Thus, innate recognition of HSV by human primary macrophages occurs via two distinct intracellular nucleotide-sensing pathways responsible for induction of IFNs and inflammatory cytokine expression, respectively.
authors.affiliation: Department of Infectious Diseases, Aarhus University Hospital Skejby, and Department of Medical Microbiology and Immunology, University of Aarhus, Brendstrupgaardsvej 100, DK-8200 Aarhus N, Denmark. jespmelc@rm.dk
authors.forename: Jesper
authors.initials: J
authors.lastname: Melchjorsen
authors.forename: Johanna
authors.initials: J
authors.lastname: Rintahaka
authors.forename: Stine
authors.initials: S
authors.lastname: Søby
authors.forename: Kristy A
authors.initials: KA
authors.lastname: Horan
authors.forename: Alina
authors.initials: A
authors.lastname: Poltajainen
authors.forename: Lars
authors.initials: L
authors.lastname: Østergaard
authors.forename: Søren R
authors.initials: SR
authors.lastname: Paludan
authors.forename: Sampsa
authors.initials: S
authors.lastname: Matikainen
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007113:Immunity, Innate; D000072640:Interferon-Induced Helicase, IFIH1; D007372:Interferons / Q000096:biosynthesis; D008264:Macrophages / Q000821:virology*; D012320:RNA Polymerase III / Q000378:metabolism; D011971:Receptors, Immunologic; D015398:Signal Transduction / Q000276:immunology; D018139:Simplexvirus / Q000276:immunology*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D016207:Cytokines; C505371:MAVS protein, human; D011971:Receptors, Immunologic; D007372:Interferons; D012320:RNA Polymerase III; C452820:RIGI protein, human; C452815:IFIH1 protein, human; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases; D000072640:Interferon-Induced Helicase, IFIH1