Inhibitors of TLR-9 act on multiple cell subsets in mouse and man in vitro and prevent death in vivo from systemic inflammation.
In parallel with the discovery of the immunostimulatory activities of CpG-containing oligodeoxynucleotides, several groups have reported specific DNA sequences that could inhibit activation by CpG-containing oligodeoxynucleotides in mouse models. We show that these inhibitory sequences, termed IRS, inhibit TLR-9-mediated activation in human as well as mouse cells. This inhibitory activity includes proliferation and IL-6 production by B cells, and IFN-alpha and IL-12 production by plasmacytoid dendritic cells. Our studies of multiple cell types in both mice and humans show the optimal IRS to contain a GGGG motif within the sequence, and the activity to require a phosphorothioate backbone. Although the GGGG motif readily itself leads to formation of a tetrameric oligodeoxynucleotide structure, inhibitory activity resides exclusively in the single-stranded form. When coinjected with a CpG oligodeoxynucleotide in vivo, IRS were shown to inhibit inflammation through a reduction in serum cytokine responses. IRS do not need to be injected at the same site to inhibit, demonstrating that rapid, systemic inhibition of TLR-9 can be readily achieved. IRS can also inhibit a complex pathological response to ISS, as shown by protection from death after massive systemic inflammation induced by a CpG-containing oligodeoxynucleotides.
Duramad
Omar
O
Dynavax Technologies, Berkeley, CA 94710, USA.
Fearon
Karen L
KL
Chang
Bonnie
B
Chan
Jean H
JH
Gregorio
Josh
J
Coffman
Robert L
RL
Barrat
Franck J
FJ
D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000494:pharmacology; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000008:administration & dosage / Q000494:pharmacology; D002469:Cell Separation; D018899:CpG Islands / Q000276:immunology; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000502:physiology; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism; D006151:Guanosine / Q000008:administration & dosage / Q000494:pharmacology; D006801:Humans; D007279:Injections, Subcutaneous; D016898:Interferon-alpha / Q000037:antagonists & inhibitors; D016753:Interleukin-10 / Q000502:physiology; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D016131:Lymphocyte Subsets / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage* / Q000494:pharmacology*; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors* / Q000502:physiology; D018805:Sepsis / Q000276:immunology / Q000401:mortality* / Q000473:pathology / Q000517:prevention & control*; D013154:Spleen / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D013873:Thionucleotides / Q000008:administration & dosage / Q000494:pharmacology; D051217:Toll-Like Receptor 9
D000276:Adjuvants, Immunologic; D000894:Anti-Inflammatory Agents, Non-Steroidal; C408982:CPG-oligonucleotide; D004268:DNA-Binding Proteins; D016898:Interferon-alpha; D009838:Oligodeoxyribonucleotides; D011956:Receptors, Cell Surface; C495371:TLR9 protein, human; D013873:Thionucleotides; C495370:Tlr9 protein, mouse; D051217:Toll-Like Receptor 9; D006151:Guanosine; D016753:Interleukin-10