title: Murine J774 macrophages recognize LPS/IFN-g, non-CpG DNA or two-CpG DNA-containing sequences as immunologically distinct.
abstract: Specific bacterial lipopolysaccharides (LPS), IFN-gamma, and unmethylated cytosine or guanosine-phosphorothioate containing DNAs (CpG) activate host immunity, influencing infectious responses. Macrophages detect, inactivate and destroy infectious particles, and synthetic CpG sequences invoke similar responses of the innate immune system. Previously, murine macrophage J774 cells treated with CpG induced the expression of nitric oxide synthase 2 (NOS2) and cyclo-oxygenase 2 (COX2) mRNA and protein. In this study murine J774 macrophages were exposed to vehicle, interferon gamma+lipopolysaccharide (IFN-g/LPS), non-CpG (SAK1), or two-CpG sequence-containing DNA (SAK2) for 0-18h and gene expression changes measured. A large number of immunostimulatory and inflammatory changes were observed. SAK2 was a stronger activator of TNFalpha- and chemokine expression-related changes than LPS/IFN-g. Up regulation included tumor necrosis factor receptor superfamily genes (TNFRSF's), IL-1 receptor signaling via stress-activated protein kinase (SAPK), NF-kappaB activation, hemopoietic maturation factors and sonic hedgehog/wingless integration site (SHH/Wnt) pathway genes. Genes of the TGF-beta pathway were down regulated. In contrast, LPS/IFN-g-treated cells showed increased levels for TGF-beta signaling genes, which may be linked to the observed up regulation of numerous collagens and down regulation of Wnt pathway genes. SAK1 produced distinct changes from LPS/IFN-g or SAK2. Therefore, J774 macrophages recognize LPS/IFN-g, non-CpG DNA or two-CpG DNA-containing sequences as immunologically distinct.
authors.affiliation: University of Tennessee Health Sciences Center, Department of Physiology, Memphis, TN 38163, USA. lcrosby@uthsc.edu
authors.forename: Lynn
authors.initials: L
authors.lastname: Crosby
authors.forename: Warren
authors.initials: W
authors.lastname: Casey
authors.forename: Kevin
authors.initials: K
authors.lastname: Morgan
authors.forename: Hong
authors.initials: H
authors.lastname: Ni
authors.forename: Lawrence
authors.initials: L
authors.lastname: Yoon
authors.forename: Marilyn
authors.initials: M
authors.lastname: Easton
authors.forename: Mary
authors.initials: M
authors.lastname: Misukonis
authors.forename: Gary
authors.initials: G
authors.lastname: Burleson
authors.forename: Dipak K
authors.initials: DK
authors.lastname: Ghosh
mesh_terms: D000818:Animals; D002460:Cell Line; D016000:Cluster Analysis; D018899:CpG Islands / Q000276:immunology*; D004247:DNA / Q000276:immunology*; D007150:Immunohistochemistry; D007371:Interferon-gamma / Q000276:immunology*; D008070:Lipopolysaccharides / Q000276:immunology*; D008264:Macrophages / Q000166:cytology / Q000276:immunology*; D051379:Mice; D009711:Nucleotides / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D008070:Lipopolysaccharides; D009711:Nucleotides; D007371:Interferon-gamma; D004247:DNA