title: Differential induction of MyD88- and TRIF-dependent pathways in equine monocytes by Toll-like receptor agonists.
abstract: Our understanding of the innate immune response in the horse has been limited by a lack of definitive data concerning cell signaling in response to microbial products. Toll-like receptors (TLRs) recognize conserved molecular motifs of microbes and elicit immune responses through their coupling with intracellular adaptor molecules, particularly MyD88 and TRIF. To provide a more definitive characterization of TLR signaling in the horse, the objectives of this study were to: (1) characterize the responses of equine monocytes to TLR ligands that signal through MyD88, TRIF or both in other species, and (2) determine the profiles of gene expression initiated utilizing these adaptor molecules. Monocytes were used to establish concentration response curves for Escherichia coli lipopolysaccharide (LPS; TLR4 ligand) and N-palmitoyl-S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-[R]-cysteinyl-[S]-seryl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysine x 3 HCl (Pam(3)CSK(4); TLR2 ligand) based on expression of procoagulant activity (PCA) and production of tumor necrosis factor-alpha (TNF-alpha); effects of polyinosine-polycytidylic acid (Poly I:C; TLR3 ligand) were determined by quantifying expression of mRNA for interferon-beta (IFN-ss). Expression of genes associated with the MyD88- (TNF-alpha, IL-1ss, IL-6 and IL-10) and TRIF-dependent pathways (IFN-ss, IP-10, RANTES and TRAF1) were measured at intervals spanning 20 h. LPS and Pam(3)CSK(4) induced significantly higher expression of TNF-alpha, IL-1ss, and IL-10 than did Poly I:C. Poly I:C induced significantly higher expression of IFN-ss, IP-10 and RANTES than did either the TLR2 or TLR4 ligands. High concentrations of E. coli LPS did not significantly increase expression of genes associated with the TRIF-dependent pathway. The results of this study suggest that equine monocytes utilize a common intracellular pathway in response to TLR2 and TLR4 ligands, but a distinct pathway in response to TLR3 ligands.
authors.affiliation: Department of Large Animal Medicine, College of Veterinary Medicine, The University of Georgia, Athens, GA 30602-7385, USA. mf239@uga.edu
authors.forename: Monica D
authors.initials: MD
authors.lastname: Figueiredo
authors.forename: Michel L
authors.initials: ML
authors.lastname: Vandenplas
authors.forename: David J
authors.initials: DJ
authors.lastname: Hurley
authors.forename: James N
authors.initials: JN
authors.lastname: Moore
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D018946:Chemokine CCL5 / Q000235:genetics; D054357:Chemokine CXCL10 / Q000235:genetics; D016207:Cytokines / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D020869:Gene Expression Profiling; D006736:Horses / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000235:genetics; D008024:Ligands; D055666:Lipopeptides / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D009000:Monocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D053594:Myeloid Differentiation Factor 88 / Q000378:metabolism*; D011070:Poly I-C / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D051193:Toll-Like Receptors / Q000819:agonists*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D018946:Chemokine CCL5; D054357:Chemokine CXCL10; D016207:Cytokines; D017931:DNA Primers; D008024:Ligands; D055666:Lipopeptides; D008070:Lipopolysaccharides; D053594:Myeloid Differentiation Factor 88; C508075:Pam(3)CSK(4) peptide; D012333:RNA, Messenger; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D011070:Poly I-C