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¹ Schering-Plough, Laboratory for Immunological Research, 69571 Dardilly, France
² Department of Microbiology, Medicine, McGill University, Montreal, Quebec H3T 1E2, Canada
³ Department of Immunobiology, Medicine, McGill University, Montreal, Quebec H3T 1E2, Canada

CORRESPONDENCE Christophe Caux: cauxc{at}lyon.fnclcc.fr

Dendritic cells (DC) produce interleukin-12 (IL-12) in response to Toll-like receptor (TLR) activation. Two major TLR signaling pathways participate in the response to pathogens: the nuclear factor-B (NF-B)–dependent pathway leading to inflammatory cytokine secretion including IL-12 and the interferon (IFN)-dependent pathway inducing type I IFN and IFN-regulated genes. Here we show that the two pathways cooperate and are likely both necessary for inducing an optimal response to pathogens. R-848/Resiquimod (TLR7 ligand in the mouse and TLR7/8 ligand in human) synergized with poly(I:C) (TLR3 ligand) or lipopolysaccharide (LPS; TLR4 ligand) in inducing high levels of bioactive IL-12p70 secretion and IFN-ß mRNA accumulation by mouse bone marrow–derived DC (BM-DC). Strikingly, IL-12p70 but not IL-12p40 secretion was strongly reduced in BM-DC from STAT1^–/– and IFNAR^–/– mice. STAT1 tyrosine-phosphorylation, IL-12p35, and IFN-ß mRNA accumulation were strongly inhibited in IFNAR^–/– BM-DC activated with the TLR ligand combinations. Similar observation were obtained in human TLR8-expressing monocyte-derived DC (moDC) using neutralizing anti-IFNAR2 antibodies, although results also pointed to a possible involvement of IFN-1 (also known as IL-29). This suggests that TLR engagement on DC induces endogenous IFNs that further synergize with the NF-B pathway for optimal IL-12p70 secretion. Moreover, analysis of interferon regulatory factors (IRF) regulation in moDC suggests a role for IRF7/8 in mediating IRF3-independent type I IFN and possibly IL-12p35 synthesis in response to TLR7/8.

Abbreviations used: CHX, cycloheximide; IRF-3, IFN regulatory factor 3; LRR, leucine-rich repeats; moDC, monocyte-derived DC; NF-B, nuclear factor-B; pDC, plasmacytoid DC; TICAM, TIR-containing adaptor molecule; TIR, Toll/IL-1 receptor; TLR, toll-like receptor; TRIF, TIR-containing adaptor inducing IFN-ß.

M. Humbert's present address is INSERM-U624, 13288 Marseille, France.

G. Trinchieri's present address is Laboratory of Parasitic Diseases, NIAID/NIH, Bethesda, MD 20892-8003.

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