title: TLR7 and TLR8 ligands and antiphospholipid antibodies show synergistic effects on the induction of IL-1beta and caspase-1 in monocytes and dendritic cells.
abstract: TLRs represent the first line of defense against invading pathogens in the innate immune system. Certain cytokines are important mediators and essentially necessary to assure an appropriately regulated immune response. Recent data gave initial evidence that IL-1beta is one of the most relevant members of these regulating cytokines. We investigated the induction of IL-1beta production in monocytes and pDCs stimulated with ligands for TLR7 and TLR8 and with antiphospholipid antibodies (aPL). Using human monocytes and pDCs for stimulation with specific TLR7 and TLR8 ligands such as resiquimod (R848) and single stranded RNA (RNA42) as well as with a human monoclonal aPL HL5B resulted in a specific upregulation of IL-1beta mRNA and protein in these cells. Determination of expression-levels using real-time RT-PCR showed significantly augmented TLR-dependent IL-1beta and caspase-1 expression. This increase could be substantially enhanced by adding the monoclonal aPL HL5B. To demonstrate the direct dependency between TLR stimulation and IL-1beta production, specific TLR inhibitors were applied and the IL-1beta and caspase-1 secretion could be explicitly decreased. The respective protein levels were determined using Western Blot, FACS analysis or ELISA assays. In conclusion we demonstrated that the downstream signaling pathway of TLR7 and TLR8 in monocytes and pDCs after stimulation with specific ligands included not only the secretion of cytokines such as TNFalpha and IL-1beta but as well the activation of necessary regulating proteins like caspase-1. APL seem to enforce this process hinting that endogenous stimulation of TLRs in the Antiphospholipid Syndrome (APS) patients resulted in enhanced secretion of proinflammatory cytokines.
authors.affiliation: Institute of Clinical Chemistry and Laboratory Medicine, Johannes Gutenberg University, Lagenbeckstr. 1, 55131 Mainz, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Hurst
authors.forename: Nadine
authors.initials: N
authors.lastname: Prinz
authors.forename: Mareike
authors.initials: M
authors.lastname: Lorenz
authors.forename: Stefan
authors.initials: S
authors.lastname: Bauer
authors.forename: Joab
authors.initials: J
authors.lastname: Chapman
authors.forename: Karl J
authors.initials: KJ
authors.lastname: Lackner
authors.forename: Philipp
authors.initials: P
authors.lastname: von Landenberg
mesh_terms: D017152:Antibodies, Antiphospholipid / Q000276:immunology* / Q000378:metabolism; D016736:Antiphospholipid Syndrome / Q000276:immunology; D020170:Caspase 1 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002469:Cell Separation; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D004790:Enzyme Induction / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D053583:Interleukin-1beta / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008024:Ligands; D008297:Male; D009000:Monocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000276:immunology; D012313:RNA / Q000276:immunology; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics / Q000276:immunology; D051199:Toll-Like Receptor 7 / Q000819:agonists / Q000235:genetics / Q000276:immunology*; D051200:Toll-Like Receptor 8 / Q000819:agonists / Q000235:genetics / Q000276:immunology*
chemical_list: D017152:Antibodies, Antiphospholipid; D007093:Imidazoles; D053583:Interleukin-1beta; D008024:Ligands; D016376:Oligonucleotides, Antisense; C495348:TLR7 protein, human; C495349:TLR8 protein, human; D051199:Toll-Like Receptor 7; D051200:Toll-Like Receptor 8; D012313:RNA; D020170:Caspase 1; C402365:resiquimod