title: Mechanisms for macrophage-mediated HIV-1 induction.
abstract: Viral latency is a long-term pathogenic condition in patients infected with HIV-1. Low but sustained virus replication in chronically infected cells can be activated by stimulation with proinflammatory cytokines such as TNF-alpha, IL-1 beta, or other host factors. However, the precise mechanism by which cellular activation induces latently infected cells to produce virions has remained unclear. In the present report, we present evidence that activation of HIV-1 replication in latently infected U1 or ACH2 cells by human macrophages is mediated by a rapid nuclear localization of NF-kappaB p50/p65 dimer with concomitant increased expression of proinflammatory cytokines. Multiplexed RT-PCR amplification of mRNA isolated from cocultures of macrophages and U1 and ACH2 cells showed significant induction of IL-1beta, IL-6, IL-8, TNF-alpha, and TGF-beta expression within 3 h of coincubation. Fixation of macrophages, U-1, or ACH2 cells with paraformaldehyde before coculture completely abrogated the induction of NF-kappaB subunits and HIV-1 replication, suggesting that cooperative interaction between the two cell types is an essential process for cellular activation. Pretreatment of macrophage-U1 or macrophage-ACH2 cocultures with neutralizing anti-TNF-alpha Ab down-regulated the replication of HIV-1. In addition, pretreatment of macrophage-U1 or macrophage-ACH2 cocultures with the NF-kappaB inhibitor (E)3-[(4-methylphenyl)sulfonyl]-2-propenenitrile (BAY 11-7082) prevented the induction of cytokine expression, indicating a pivotal role of NF-kappaB-mediated signaling in the reactivation of HIV-1 in latently infected cells by macrophages. These results provide a mechanism by which macrophages induce HIV-1 replication in latently infected cells.
authors.affiliation: Immunopathogenesis Section, Laboratory of Molecular Virology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, USA. devadas@cber.fda.gov
authors.forename: Krishnakumar
authors.initials: K
authors.lastname: Devadas
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Hardegen
authors.forename: Larry M
authors.initials: LM
authors.lastname: Wahl
authors.forename: Indira K
authors.initials: IK
authors.lastname: Hewlett
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Clouse
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Yamada
authors.forename: Subhash
authors.initials: S
authors.lastname: Dhawan
mesh_terms: D002450:Cell Communication / Q000276:immunology; D002460:Cell Line; D018920:Coculture Techniques; D016207:Cytokines / Q000096:biosynthesis; D015497:HIV-1 / Q000276:immunology / Q000502:physiology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000821:virology; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism / Q000821:virology*; D009000:Monocytes / Q000276:immunology / Q000821:virology; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D009570:Nitriles / Q000494:pharmacology; D013450:Sulfones / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology; D020298:U937 Cells; D014775:Virus Activation / Q000276:immunology*; D017735:Virus Latency / Q000276:immunology; D014779:Virus Replication / Q000276:immunology
chemical_list: C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D016207:Cytokines; D016328:NF-kappa B; D009570:Nitriles; D013450:Sulfones; D014409:Tumor Necrosis Factor-alpha