title: A role for microRNA-155 modulation in the anti-HIV-1 effects of Toll-like receptor 3 stimulation in macrophages.
abstract: HIV-1 infection of macrophages plays a key role in viral pathogenesis and progression to AIDS. Polyinosine-polycytidylic acid (poly(I:C); a synthetic analog of dsRNA) and bacterial lipopolysaccharide (LPS), the ligands for Toll-like receptors (TLR) TLR3 and TLR4, respectively, are known to decrease HIV-1 infection in monocyte-derived macrophages (MDMs), but the mechanism(s) are incompletely understood. We found that poly(I:C)- and LPS-stimulation of MDMs abrogated infection by CCR5-using, macrophage-tropic HIV-1, and by vesicular stomatitis virus glycoprotein-pseudotyped HIV-1 virions, while TLR2, TLR7 or TLR9 agonists only partially reduced infection to varying extent. Suppression of infection, or lack thereof, did not correlate with differential effects on CD4 or CCR5 expression, type I interferon induction, or production of pro-inflammatory cytokines or β-chemokines. Integrated pro-viruses were readily detected in unstimulated, TLR7- and TLR9-stimulated cells, but not in TLR3- or TLR4-stimulated MDMs, suggesting the alteration of post-entry, pre-integration event(s). Using microarray analysis and quantitative reverse transcription (RT)-PCR, we found increased microRNA (miR)-155 levels in MDMs upon TLR3/4- but not TLR7-stimulation, and a miR-155 specific inhibitor (but not a scrambled control) partially restored infectivity in poly(I:C)-stimulated MDMs. Ectopic miR-155 expression remarkably diminished HIV-1 infection in primary MDMs and cell lines. Furthermore, poly(I:C)-stimulation and ectopic miR-155 expression did not alter detection of early viral RT products, but both resulted in an accumulation of late RT products and in undetectable or extremely low levels of integrated pro-viruses and 2-LTR circles. Reduced mRNA and protein levels of several HIV-1 dependency factors involved in trafficking and/or nuclear import of pre-integration complexes (ADAM10, TNPO3, Nup153, LEDGF/p75) were found in poly(I:C)-stimulated and miR-155-transfected MDMs, and a reporter assay suggested they are authentic miR-155 targets. Our findings provide evidence that miR-155 exerts an anti-HIV-1 effect by targeting several HIV-1 dependency factors involved in post-entry, pre-integration events, leading to severely diminished HIV-1 infection.
authors.affiliation: Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, Pennsylvania, USA.
authors.forename: Gokul
authors.initials: G
authors.lastname: Swaminathan
authors.forename: Fiorella
authors.initials: F
authors.lastname: Rossi
authors.forename: Luz-Jeannette
authors.initials: LJ
authors.lastname: Sierra
authors.forename: Archana
authors.initials: A
authors.lastname: Gupta
authors.forename: Sonia
authors.initials: S
authors.lastname: Navas-Martín
authors.forename: Julio
authors.initials: J
authors.lastname: Martín-García
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism; D000072197:ADAM10 Protein; D053829:Amyloid Precursor Protein Secretases / Q000235:genetics / Q000378:metabolism; D015704:CD4 Antigens / Q000096:biosynthesis; D002478:Cells, Cultured; D019742:Chemokines, CC / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D057809:HEK293 Cells; D015497:HIV-1 / Q000276:immunology / Q000502:physiology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D007370:Interferon Type I / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000378:metabolism / Q000494:pharmacology; D008264:Macrophages / Q000276:immunology* / Q000821:virology*; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism; D011070:Poly I-C / Q000378:metabolism / Q000494:pharmacology; D034622:RNA Interference; D034741:RNA, Small Interfering; D019713:Receptors, CCR5 / Q000096:biosynthesis; D051195:Toll-Like Receptor 2 / Q000276:immunology; D051196:Toll-Like Receptor 3 / Q000276:immunology*; D051197:Toll-Like Receptor 4 / Q000276:immunology; D051199:Toll-Like Receptor 7 / Q000276:immunology; D051217:Toll-Like Receptor 9 / Q000276:immunology; D014759:Viral Envelope Proteins
chemical_list: D015704:CD4 Antigens; D019742:Chemokines, CC; D016207:Cytokines; C028391:G protein, vesicular stomatitis virus; D036341:Intercellular Signaling Peptides and Proteins; D007370:Interferon Type I; D008070:Lipopolysaccharides; C524942:MIRN155 microRNA, human; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D035683:MicroRNAs; C079377:NUP153 protein, human; D028861:Nuclear Pore Complex Proteins; D034741:RNA, Small Interfering; D019713:Receptors, CCR5; C495342:TLR2 protein, human; C495344:TLR3 protein, human; C495345:TLR4 protein, human; C495348:TLR7 protein, human; C495371:TLR9 protein, human; D051195:Toll-Like Receptor 2; D051196:Toll-Like Receptor 3; D051197:Toll-Like Receptor 4; D051199:Toll-Like Receptor 7; D051217:Toll-Like Receptor 9; D014759:Viral Envelope Proteins; C119380:lens epithelium-derived growth factor; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human; D011070:Poly I-C