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CUTTING EDGE |
Departments of
*
Microbiology, Molecular Genetics, and Immunology and
Biochemistry, University of Kansas Medical Center, Kansas City, KS 66160;
U.S. Human Health Division, Merck & Company Inc., West Point, PA 19044;
§
Department of Basic Medical Sciences, University of Missouri, Kansas City, MO 64108; and
¶
Saint Luke’s Hospital of Kansas City, Kansas City, MO 64111
LPS is well recognized for its potent capacity to activate mouse macrophages to produce NO, an important inflammatory mediator in innate host defense. We demonstrate here that, although inducing little NO alone, DNA from both Gram-negative and Gram-positive bacteria synergizes with subthreshold concentrations of LPS (0.3 ng/ml) to induce NO in cultures of RAW 264.7 macrophages. The effects of the DNA are mimicked by synthetic CpG-containing oligodeoxynucleotides but not by non-CpG-containing oligodeoxynucleotides. This synergistic activity is not inhibited by neutralizing Abs against IFN. Preincubation of macrophages with DNA for 8–24 h suppresses subsequent synergistic macrophage responses to DNA/LPS, whereas prolonged pretreatment with LPS enhances synergy. RT-PCR analysis indicates that the mRNA levels of the inducible NO synthase gene are also coordinately suppressed or induced. These findings indicate that temporally controlled, synergistic interactions exist between microbial DNA and LPS in the induction of macrophage NO via enhanced inducible NO synthase gene expression.
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