Abstract
Gram-positive bacteria induce the production of several cytokines in human leukocytes. The molecular mechanisms involved in Gram-positive bacteria-induced cytokine production have been poorly characterized. In this work we demonstrate that both nonpathogenic Lactobacillus rhamnosus GG and pathogenic Streptococcus pyogenes (group A streptococci) induce NF-kappa B and STAT DNA-binding activity in human primary macrophages as analyzed by EMSA. NF-kappa B activation was rapid and was not inhibited by a protein synthesis inhibitor cycloheximide, suggesting that these bacteria could directly activate NF-kappa B. STAT1, STAT3, and IFN regulatory factor-1 DNA binding was induced by both bacteria with delayed kinetics compared with NF-kappa B. In addition, streptococci induced the formation of IFN-alpha-specific transcription factor complex and IFN-stimulated gene factor-3 (ISGF3). STAT1 and STAT3 activation and ISGF3 complex formation were inhibited by cycloheximide or by neutralization with IFN-alpha/beta-specific Abs. Streptococci were more potent than lactobacilli in inducing STAT1, ISGF3, and IFN regulatory factor-1 DNA binding. Accordingly, only streptococci induced IFN-alpha production. The activation of the IFN-alpha signaling pathway by streptococci could play a role in the pathogenesis of these bacteria. These results indicate that extracellular Gram-positive bacteria activate transcription factors involved in cytokine signaling by two mechanisms: directly, leading to NF-kappa B activation, and indirectly via cytokines, leading to STAT activation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Binding Sites / immunology
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Child
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DNA / metabolism
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DNA-Binding Proteins / antagonists & inhibitors
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DNA-Binding Proteins / biosynthesis
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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DNA-Binding Proteins / physiology
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Humans
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Immune Sera / pharmacology
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Interferon Regulatory Factor-1
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Interferon-Stimulated Gene Factor 3
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Interferon-Stimulated Gene Factor 3, gamma Subunit
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Interferon-alpha / biosynthesis
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Interferon-alpha / immunology
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Interleukin-6 / biosynthesis
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Lactobacillus / drug effects
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Lactobacillus / immunology*
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Macrophages / drug effects
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Macrophages / immunology*
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Macrophages / metabolism
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Macrophages / microbiology*
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NF-kappa B / metabolism*
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NF-kappa B / physiology
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Phosphoproteins / biosynthesis
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Phosphoproteins / genetics
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Phosphoproteins / metabolism
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Protein Synthesis Inhibitors / pharmacology
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RNA, Messenger / biosynthesis
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STAT1 Transcription Factor
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STAT3 Transcription Factor
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Signal Transduction / drug effects
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Signal Transduction / immunology*
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Streptococcus pyogenes / drug effects
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Streptococcus pyogenes / immunology*
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Trans-Activators / antagonists & inhibitors
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Trans-Activators / metabolism*
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Trans-Activators / physiology
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Transcription Factors / metabolism
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Tumor Necrosis Factor-alpha / biosynthesis
Substances
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DNA-Binding Proteins
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IRF1 protein, human
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IRF9 protein, human
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Immune Sera
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Interferon Regulatory Factor-1
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Interferon-Stimulated Gene Factor 3
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Interferon-Stimulated Gene Factor 3, gamma Subunit
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Interferon-alpha
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Interleukin-6
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NF-kappa B
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Phosphoproteins
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Protein Synthesis Inhibitors
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RNA, Messenger
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STAT1 Transcription Factor
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STAT1 protein, human
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STAT3 Transcription Factor
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STAT3 protein, human
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Trans-Activators
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Transcription Factors
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Tumor Necrosis Factor-alpha
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DNA