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Autocrine–paracrine prostaglandin E2 signaling restricts TLR4 internalization and TRIF signaling

Nature Immunology volume 19, pages 1309–1318 (2018)Cite this article

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Abstract

The unique cell biology of Toll-like receptor 4 (TLR4) allows it to initiate two signal-transduction cascades: a signal dependent on the adaptors TIRAP (Mal) and MyD88 that begins at the cell surface and regulates proinflammatory cytokines, and a signal dependent on the adaptors TRAM and TRIF that begins in the endosomes and drives the production of type I interferons. Negative feedback circuits to limit TLR4 signals from both locations are necessary to balance the inflammatory response. We describe a negative feedback loop driven by autocrine–paracrine prostaglandin E2 (PGE2) and the PGE2 receptor EP4 that restricted TRIF-dependent signals and the induction of interferon-β through the regulation of TLR4 trafficking. Inhibition of PGE2 production or antagonism of EP4 increased the rate at which TLR4 translocated to endosomes and amplified TRIF-dependent activation of the transcription factor IRF3 and caspase-8. This PGE2-driven mechanism restricted TLR4–TRIF signaling in vitro after infection of macrophages by the Gram-negative pathogens Escherichia coli or Citrobacter rodentium and protected mice against mortality induced by Salmonella enteritidis serovar Typhimurium. Thus, PGE2 restricted TLR4–TRIF signaling specifically in response to lipopolysaccharide.

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Fig. 1: Rapid LPS-dependent PGE2 limits TLR4-mediated production of IFN-β.
Fig. 2: PGE2 activation of EP4 receptor restricts IRF3 activation.
Fig. 3: The PGE2–EP4 axis regulates TLR4 signaling via TRIF.
Fig. 4: PGE2–EP4 restricts the LPS-induced, TRIF-dependent activation of caspase-8 and atypical processing of IL-1β.
Fig. 5: cAMP-dependent effector molecules are negative regulators of the TLR4–TRIF pathway.
Fig. 6: PGE2 restricts IRF3 activation and IFN-β production in response to Gram negative pathogens in vitro and in vivo.

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Data availability

The data that support the findings of this study are available from the corresponding author upon request. Microarray data associated with Fig. 1 have been deposited win the GEO database with accession code GSE111826.

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Acknowledgements

We thank E. Hewlett (University of Virginia) for purified AC from B. pertussis; L. Wahl (National Institute of Dental and Craniofacial Research, US National Institutes of Health) for blood from healthy human volunteers at the Department of Transfusion Medicine of the US National Institutes of Health; R. Ernst (University of Maryland, Baltimore) for Salmonella Typhimurium strain SL1344; J.B. Kaper (University of Maryland, Baltimore) for enterohemorrhagic and enteropathogenic E. coli and C. rodentium; R. Rajaiah for technical assistance with the TLR4-internalization assay; and J.C. Blanco and D. Prantner for critique of this manuscript. Flow cytometry and microarray analyses were performed at the Center for Innovative Biomedical Resources at the University of Maryland, School of Medicine. Supported by the US National Institutes of Health (AI123371 and AI125215 to S.N.V.; and AR061491 to B.K.).

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  1. Department of Microbiology and Immunology, University of Maryland, Baltimore, School of Medicine, Baltimore, MD, USA

    Darren J. Perkins, Katharina Richard, Anne-Marie Hansen, Wendy Lai, Shreeram Nallar & Stefanie N. Vogel

  2. Department of Genetics, UNC School of Medicine, Chapel Hill, NC, USA

    Beverly Koller

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Contributions

D.J.P. performed the majority of the experiments; K.R. performed TLR4-translocation experiments; A.-M.H. performed infection with E. coli and C. rodentium; W.L. performed ELISA of IFN-β and analyzed the data; S.N. bred and genotyped all mice with targeted mutations; B.K. developed, bred, genotyped and isolated femurs from Ptger4–/– mice and their littermates; and D.J.P. and S.N.V. conceived of the study, designed the experiments and wrote the manuscript.

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Correspondence to Darren J. Perkins or Stefanie N. Vogel.

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Perkins, D.J., Richard, K., Hansen, AM. et al. Autocrine–paracrine prostaglandin E2 signaling restricts TLR4 internalization and TRIF signaling. Nat Immunol 19, 1309–1318 (2018). https://doi.org/10.1038/s41590-018-0243-7

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