Human TLR7 Or TLR8 Independently Confer Responsiveness to the Antiviral Compound R-848
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CORRESPONDENCE bers of this family (TLR1 to In the quest to identify ligands for TLRs, Human TLR7 or TLR8 TLR10) have been reported to we also screened immunostimulatory synthet- date. TLR2, TLR4 and TLR5 ic compounds for their potential to activate are crucial for the recognition HEK293 cells that were transiently transfect- independently confer of peptidoglycan, lipopolysac- ed with TLR cDNAs and a NF-κB luciferase charide and flagellin, respec- reporter plasmid. We found that R-848 responsiveness to the tively3–5. TLR6 associates with induced NF-κB activation in HEK293 cells TLR2 and recognizes lipopro- transfected with human TLR8 (GenBank antiviral compound teins from mycoplasma6. TLR9 accession number AF245703) in a dose- detects bacterial DNA contain- dependent manner (Fig. 1). In accordance ing unmethylated CpG motifs with the findings of Hemmi et al., human and R-848 and TLR3 activates immune murine TLR7 (GenBank accession numbers cells in response to double- AF240467 and AY035889) also mediate 1, 2, 1 MARION JURK *, FLORIAN HEIL *, JÖRG VOLLMER , stranded RNA7–9. The natural R-848 recognition (Fig. 1)10. TLR7 showed a CHRISTIAN SCHETTER1,ARTHUR M. KRIEG1,HERMANN ligands for TLR1, TLR7, higher sensitivity to R-848, but TLR8 was WAGNER2,GRAYSON LIPFORD1 AND STEFAN BAUER2 TLR8 and TLR10 are not able to induce NF-κB more effectively than known, although a synthetic TLR7 when higher concentrations of R-848 1Coley Pharmaceutical GmbH, Elisabeth-Selbert-Strasse 9, 40764 Langenfeld, Germany and Coley Pharmaceutical Group, Inc., 93 Worcester compound with antiviral activi- were used (Fig. 1). In contrast, HEK293 cells St.,Wellesley, MA 02481, USA. 2Institute for Medical Microbiology, Hygiene ty has now been described as a transiently transfected with murine TLR8 and Immunology,Trogerstrasse 9, 81675 Munich, Germany. *These ligand for TLR710. (GenBank accession number AY035890) did authors contributed equally to this work. ([email protected]) Hemmi et al. reported in not activate NF-κB after stimulation with Nature Immunology that, in R-848 (data not shown), which suggests that Toll-like receptors (TLRs) play an important experiments that used gene-deficient mice, the TLR8 is nonfunctional in mice. This mute role in the innate immune response to antiviral imidazoquinoline resiquimod (R-848) phenotype is in accordance with the observa- pathogens1. TLRs detect PAMPs (pathogen- activates immune cells via the TLR7 MyD88- tion that TLR7-deficient mice do not respond associated molecular patterns) and stimulate dependent signaling pathway10. They showed to R-848, even though TLR8 is present10. immune cells via the MyD88-dependent inter- that macrophages from MyD88- and TLR7- These results show that both human TLR7 leukin 1 receptor (IL-1R)–TLR signaling deficient mice do not respond to R-848 stimu- and TLR8 can independently mediate recogni- pathway, which leads to activation of the tran- lation, whereas macrophages from wild-type tion of the same antiviral compound, imidazo- scription factor NF-κB2. In humans, ten mem- mice strongly induce the transcription factor quinoline R-848, which suggests a possible NF-κB and the secretion of proinflammatory redundancy among these receptors. The differ- cytokines, such as tumor necrosis factor-α, as ential dose response observed between these © http://immunol.nature.com Group 2002 Nature Publishing 20 Human TLR8 well as the regulatory cytokine IL-12. In addi- receptors may allow fine-tuning of the 15 tion, Hemmi et al. showed, by genetic com- immune response to high or low concentra- 10 plementation in HEK293 cells, that human tions of this molecule as well as other ligands. 5 TLR7 confers responsiveness to R-84810. Further studies will be necessary to identify 0 the natural ligands for both receptors and clar- Human TLR7 ify the status and function of murine TLR8. 5 Figure 1. R-848 induces NF-κB activation via B induction κ TLR8 or TLR7. HEK293 cells were transfected by elec- troporation with vectors expressing human TLR8,human TLR7 or murine TLR7 and a NF-kB luciferase reporter 1. Medzhitov, R. Nature Rev. Immunol. 1, 135–145 (2001). Fold NF- 0 2. Medzhitov, R. et al. Mol. Cell 2, 253–258 (1998). plasmid. Sixteen hours after transfection, cells were stim- 3. Takeuchi, O. et al. Immunity 11, 443–451 (1999). Murine TLR7 ulated with R-848 (commercially synthesized by 4. Poltorak,A. et al. Science 282, 2085–2088 (1998). 5 GLSynthesis,Worcester,MA) at the indicated concentra- 5. Hayashi, F. et al. Nature 410, 1099–1103 (2001). tions for a further 7 h.TLR2-,TLR3- and TLR4-transfect- 6. Ozinsky,A. et al. Proc. Natl.Acad. Sci. USA 97, 13766–13771 (2000). 7. Hemmi, H. et al. Nature 408, 740–745 (2000). ed cells did not induce NF-kB activation after stimulation 8. Bauer, S. et al. Proc. Natl.Acad. Sci. USA 98, 9237–9242 (2001). 0 with R-848; a negative result was also scored for murine 9. Alexopoulou, L., Holt, H. C., Medzhitov, R. & Flavell, R.A. Nature -3 10 413, 732–738 (2001). R-848 0 10 10-2 10-1 1 10 TLR8 (data not shown ). Data are mean±s.d. from one (µg/ml) representative of three independent experiments. 10. Hemmi, H. et al. Nature Immunol. 3, 196–200 (2002). http://immunol.nature.com • june 2002 • volume 3 no 6 • nature immunology 499.