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The Wistar Institute of Anatomy and Biology, Philadelphia, PA 19104
Exogenously added IL-10 rapidly inhibited Staphylococcus
aureus- or LPS-induced cytokine mRNA expression in human PBMCs
and monocytes, with a maximal effect observed when IL-10 was added from
20 h before until 1 h after the addition of the inducers.
Nuclear run-on assays revealed that the inhibition of IL-12 p40, IL-12
p35, and TNF-
was at the gene transcriptional level and that the
addition of IL-10 to S. aureus- or LPS-treated PBMCs did
not affect mRNA stability. The inhibitory activity of IL-10 was
abrogated by cycloheximide (CHX), suggesting the involvement of a newly
synthesized protein(s). The addition of CHX at 2 h before S.
aureus or LPS also inhibited the accumulation of IL-12 p40 mRNA,
but did not inhibit IL-12 p35 and TNF- mRNA. This finding suggests
that p40 transcription is regulated through a de novo synthesized
protein factor(s), whereas the addition of CHX at 2 h after
S. aureus activation caused superinduction of the
IL-12 p40, IL-12 p35, and TNF-
genes. These results indicate that in human monocytes, the mechanism(s)
of IL-10 suppression of both IL-12 p40 and IL-12
p35 genes is primarily seen at the transcriptional level, and
that the induction of the IL-12 p40 and p35
genes have different requirements for de novo protein synthesis.
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