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Through a Post-Transcriptional Mechanism1
*
Department of Basic Medical Sciences, University of Missouri, and
Saint Luke’s Hospital, Kansas City, MO 64111
LPS is well recognized for its potent capacity to activate mouse
macrophages to produce TNF-
, an important inflammatory mediator in
bacterial infection-related diseases such as septic shock. We
demonstrate here that while inducing only low levels of TNF- alone,
DNA from both Gram-negative and Gram-positive bacteria synergizes with
subthreshold concentrations of LPS (0.3 ng/ml) to induce TNF- in the
RAW 264.7 macrophage-like cell line. The bacterial DNA effects are
mimicked by synthetic CpG-containing oligodeoxynucleotides, but not
non-CpG-containing oligodeoxynucleotides. Pretreatment of macrophages
with either DNA for 2–8 h inhibits macrophage TNF- production in
responses to DNA/LPS. However, when pretreatment was extended to
24 h, DNA/LPS synergy on TNF- is further enhanced. RT-PCR
analysis indicates that mRNA levels of the TNF- gene, however, are
not synergistically induced by bacterial DNA and LPS. Analyses of the
half-life of TNF- mRNA indicate that TNF- message has a longer
half-life in bacterial DNA- and LPS-treated macrophages than that in
bacterial DNA- or LPS-treated macrophages. These findings indicate that
the temporally controlled, synergistic induction of TNF- by
bacterial DNA and LPS is not mediated at the transcriptional level.
Instead, this synergy may occur via a post-transcriptional
mechanism.
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