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Notch–RBP-J signaling regulates the transcription factor IRF8 to promote inflammatory macrophage polarization

Nature Immunology volume 13, pages 642–650 (2012)Cite this article

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Abstract

Emerging concepts suggest that the functional phenotype of macrophages is regulated by transcription factors that define alternative activation states. We found that RBP-J, the main nuclear transducer of signaling via Notch receptors, augmented Toll-like receptor 4 (TLR4)-induced expression of key mediators of classically activated M1 macrophages and thus of innate immune responses to Listeria monocytogenes. Notch–RBP-J signaling controlled expression of the transcription factor IRF8 that induced downstream M1 macrophage–associated genes. RBP-J promoted the synthesis of IRF8 protein by selectively augmenting kinase IRAK2–dependent signaling via TLR4 to the kinase MNK1 and downstream translation-initiation control through eIF4E. Our results define a signaling network in which signaling via Notch–RBP-J and TLRs is integrated at the level of synthesis of IRF8 protein and identify a mechanism by which heterologous signaling pathways can regulate the TLR-induced inflammatory polarization of macrophages.

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Figure 1: RBP-J controls the expression of prototypical M1 macrophage–associated genes.
Figure 2: Induction of RBP-J-dependent M1 macrophage–associated genes requires canonical Notch signaling.
Figure 3: RBP-J controls the expression and function of IRF8.
Figure 4: IRF8 mediates RBP-J-dependent activation of the expression of M1 macrophage–associated genes.
Figure 5: RBP-J promotes the synthesis of IRF8 protein.
Figure 6: RBP-J augments the TLR4-induced activation of the MAPK-MNK1-eIF4E pathway.
Figure 7: Notch–RBP-J signaling regulates the expression of IRAK2 protein.
Figure 8: The MAPK-MNK1-eIF4E axis promotes the synthesis of IRF8 and the expression of M1 macrophage–associated genes.

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Acknowledgements

We thank T. Honjo (Kyoto University) for Rbpjflox/flox mice; T. Gridley (Maine Medical Center Research Institute) for Notch1+/− mice; R. Kopan (Washington University) for NICD1 expression plasmids; S. Smale (University of California, Los Angeles) for Il12b promoter reporter constructs; M. Takami (Showa University) for the IRF8 expression construct; S. Akira (Osaka University) for IRAK2 retroviral constructs; J.A. Cooper (Fred Hutchinson Cancer Research Center) for MNK1 expression plasmids; E. Kieff (Harvard Medical School); J.C. Aster (Harvard Medical School) for anti-RBP-J rabbit serum; E.G. Pamer for discussions about the L. monocytogenes infection experiments; and K. Au for technical assistance. Supported by the American College of Rheumatology (X.H.) and the US National Institutes of Health (L.B.I. and X.H.).

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Author notes

  1. Haixia Xu, Jimmy Zhu and Sinead Smith: These authors contributed equally to this work.

Authors and Affiliations

  1. Arthritis and Tissue Degeneration Program, Hospital for Special Surgery, New York, New York, USA

    Haixia Xu, Jimmy Zhu, Sinead Smith, Baohong Zhao, Allen Y Chung, Carl P Blobel, Lionel B Ivashkiv & Xiaoyu Hu

  2. Department of Endocrinology, the third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China

    Haixia Xu

  3. Graduate Program in Immunology and Microbial Pathogenesis, Weill Cornell Graduate School of Medical Sciences, New York, New York, USA

    Julia Foldi, Chao Shi & Lionel B Ivashkiv

  4. Herbert Irving Comprehensive Cancer Center, Columbia University, New York, New York, USA

    Hasina Outtz & Jan Kitajewski

  5. Memorial Sloan Kettering Cancer Center, New York, New York, USA

    Chao Shi & Yueming Li

  6. Biochemisches Institute, Christian Albrechts Universität Kiel, Kiel, Germany

    Silvio Weber & Paul Saftig

  7. National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA

    Keiko Ozato

  8. Department of Medicine, Weill Cornell Medical College, New York, New York, USA

    Xiaoyu Hu

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Contributions

H.X., J.Z., J.F., A.Y.C. and C.S. did experiments and analyzed data; S.S. did experiments, analyzed data and prepared the manuscript. B.Z. generated NICD1M mice, provided the IRF8-expressing retroviral vector and assisted with the experiments with Irf8−/− mice; H.O. and J.K. provided Notch1+/− mice; S.W. and P.S. provided ADAM10-deficient mice; Y.L. provided GSI34; K.O. provided Irf8−/− mice; C.P.B. provided mice with loxP-flanked Adam17 alleles and advice about experiments; L.B.I. provided advice about experiments and contributed to manuscript preparation; and X.H. designed research, supervised experiments and prepared the manuscript.

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Correspondence to Xiaoyu Hu.

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Xu, H., Zhu, J., Smith, S. et al. Notch–RBP-J signaling regulates the transcription factor IRF8 to promote inflammatory macrophage polarization. Nat Immunol 13, 642–650 (2012). https://doi.org/10.1038/ni.2304

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