The newly identified CpG-N ODN208 protects mice from challenge with CpG-S ODN by decreasing TNF-alpha release.
Administration of an excess of oligodeoxynucleotides containing immunostimulatory CpG motifs (CpG-S ODNs) may induce systemic inflammatory response syndrome (SIRS) and sepsis. Therefore, it is important to develop neutralizing CpG ODNs (CpG-N ODNs), which can be used to reduce the release of cytokines induced by the presence of CpG-S ODNs. In the present study, CpG-N ODN208 (5'-TGCCGCGGCAGA-3'), a neutralizing twelve-oligodeoxynucleotide molecule recently identified in our laboratory, inhibited TNF-alpha release from human peripheral blood mononuclear cells (hPBMCs) and murine RAW264.7 cells induced by CpG-S ODN exposure in a dose- and time-dependent manner. Flow cytometry revealed that CpG-N ODN208 decreased cell-surface binding and internalization of 6-FAM-CpG-S ODN. However, the decreased cell-surface binding and internalization of CpG-S ODN could not completely account for the decreased TNF-alpha release. RT-PCR experiments revealed that CpG-N ODN treatment could down-regulate the CpG-S ODN-induced upregulation of Toll-like receptor 9 (TLR9) mRNA expression. This finding suggested that the decreased cytokine release following CpG-N ODN treatment might be related to decreased TLR9 mRNA expression. In in vivo experiments, no protection was found when the ratio of CpG-N ODN to CpG-S ODN delivered to mice was 3:1. However, at a 5:1 ratio, CpG-N ODN208 could protect mice from an ordinarily lethal dose of CpG-S ODN. Furthermore, we found that CpG-N ODN208 treatment decreased serum TNF-alpha levels in mice injected with sublethal doses of CpG-S ODN whether the CpG-N ODN208 was added prior to or concurrent with the CpG-S ODN. Our results demonstrated that CpG-N ODN-mediated protection against a lethal challenge by CpG-S ODN was associated with the reduction of TNF-alpha release.
Wang
Liangxi
L
Department of Pharmacology, College of Medicine, The Third Military Medical University, Chongqing 400038, PR China.
Jiang
Weiwei
W
Ding
Guofu
G
Cao
Hongwei
H
Lu
Yongling
Y
Luo
Ping
P
Zhou
Hong
H
Zheng
Jiang
J
D000256:Adenoviridae / Q000737:chemistry; D000818:Animals; D002469:Cell Separation; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004279:DNA, Viral / Q000737:chemistry; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D007202:Indicators and Reagents; D051379:Mice; D008815:Mice, Inbred Strains; D009000:Monocytes / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000037:antagonists & inhibitors* / Q000494:pharmacology / Q000633:toxicity; D012333:RNA, Messenger / Q000096:biosynthesis; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018805:Sepsis / Q000188:drug therapy / Q000473:pathology; D051217:Toll-Like Receptor 9 / Q000096:biosynthesis; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000378:metabolism
C520220:CpG-N ODN208; D004279:DNA, Viral; D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D051217:Toll-Like Receptor 9; D014409:Tumor Necrosis Factor-alpha