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* Children’s Foundation Research Center at Le Bonheur Children’s Hospital, and Department of Pediatrics, University of Tennessee Health Science Center, Memphis, TN 38103;
Department of Molecular Sciences, University of Tennessee Health Science Center, Memphis, TN 38163;
Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN 46202; and
Walther Oncology Center, Walther Cancer Institute, Indianapolis, IN 46208
Exposure of macrophages to LPS induces a state of
hyporesponsiveness to subsequent challenge with LPS. It has not been
known whether previous exposure to CpG DNA induces a similar
suppressive response to subsequent stimulation with CpG DNA. In the
present study, we demonstrate that pretreatment with CpG DNA induces
suppression of cytokine release in a murine macrophage-like cell
RAW264.7 in response to subsequent challenge by CpG DNA. Additionally,
CpG DNA-mediated activation of mitogen-activated protein kinases,
including c-Jun NH2-terminal kinase, extracellular
signal-regulated kinase, and p38, and activation of transcription
factors AP-1, CREB, NF-
B, and STAT1 are greatly suppressed in the
cells pre-exposed to CpG DNA. Pretreatment with CpG DNA also partially
inhibited LPS-mediated production of cytokines and activation of
mitogen-activated protein kinases and transcription factors. Neither
LPS nor CpG DNA treatment inhibited Toll-like receptor 4, MD2,
Toll-like receptor 9, myeloid differentiation factor 88, Toll/IL-1R
domain-containing adaptor protein, Tollip, and TNF-
receptor-associated factor 6 expression. Interestingly, CpG DNA or LPS
stimulation led to the inhibition of IL-1R-associated kinase
expression. These results indicate that CpG DNA-induced refractory of
RAW264.7 cells may be, at least in part, due to suppressed
IL-1R-associated kinase expression.
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